The Effects Of N-Acetylcysteine on Asprosine Immunoreactivity in A Rat Ovarian Torsion-Detorsion Model

The Effects Of N-Acetylcysteine on Asprosine Immunoreactivity in A Rat Ovarian Torsion-Detorsion Model | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article The Effects Of N-Acetylcysteine on Asprosine Immunoreactivity in A Rat Ovarian Torsion-Detorsion Model Gülay BULU, Alpaslan AKYOL, Tuncay KULOĞLU, Aykut BULU, Cengiz ŞANLI This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6623339/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Objective: Ovarian torsion is a life-threatening gynecological emergency that often necessitates surgical intervention. This study aimed to evaluate the effects of N-acetylcysteine (NAC), a potent antioxidant, on ovarian reserve and asprosin levels in a rat model of torsion-detorsion. Materials and Methods: Thirty-five Wistar albino rats were randomly assigned to five groups: Group I (Control), Group II (Sham), Group III (NAC), Group IV (Torsion/Detorsion - T/D), and Group V (Torsion/Detorsion + NAC). Ovarian and blood samples were collected to assess asprosin immunoreactivity in tissue, serum asprosin levels, anti-Müllerian hormone (AMH) concentrations, and total oxidant status (TOS). Histopathological changes in ovarian tissues were evaluated using immunohistochemical techniques. Results: Serum TOS levels were significantly elevated in the T/D group (p = 0.016) compared to the control group, and significantly decreased in the T/D + NAC group (p = 0.032) compared to the T/D group. Serum asprosin levels were significantly lower in the NAC (p = 0.002), T/D (p = 0.003), and T/D + NAC (p = 0.010) groups compared to the control (overall p = 0.016). However, asprosin levels were significantly higher in the T/D + NAC group than in the T/D group (p = 0.008). In ovarian tissue, asprosin immunoreactivity was significantly reduced in the T/D group compared to the control (p = 0.002), but increased in the T/D + NAC group compared to the T/D group (p = 0.002). No statistically significant differences were observed in AMH levels among the groups (p > 0.05). Conclusion: N-acetylcysteine (NAC) effectively reduced oxidative stress and prevented degenerative changes associated with ovarian torsion-detorsion injury. Asprosin levels appeared to reflect ischemia-reperfusion injury, decreasing with torsion and partially recovering with NAC treatment. Ovarian torsion-detorsion asprosin N-acetylcysteine ischemia-reperfusion immunohistochemistry rat model Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Introduction Ovarian torsion, defined as the complete or partial rotation of the ovary, fallopian tube, or both around their vascular axis, is a gynecological emergency with a prevalence of 2.7%. 1 It can affect women of all ages, and delayed diagnosis can negatively impact ovarian reserve. 2 – 3 Common causes include ovarian masses and ovulation induction. 4 However, the exact mechanisms leading to necrosis after torsion—such as the degree and duration of rotation—remain unclear. 5 This is because necrosis may not occur even in the presence of vascular pedicle twisting, whereas infarction and necrosis can sometimes develop due to venous congestion and edema despite preserved arterial blood flow. 5 Excluding prolonged ischemia, many torsion cases have shown preserved ovarian function. 6 – 8 Therefore, early detection and prompt detorsion are the primary goals in the diagnosis and management of ovarian torsion. 9 However, the reperfusion process following detorsion can induce oxidative stress through the activation of free oxygen radicals and inflammatory cells. 10 Reperfusion is believed to be a major mechanism of tissue damage, independent of ischemia itself. 11 Although numerous studies have investigated ovarian torsion, the effects of reperfusion on ovarian reserve and energy metabolism remain insufficiently understood. Anti-Mullerian Hormone (AMH) and antral follicle count are widely used parameters to assess ovarian reserve. 12 – 13 AMH is produced by growing follicles and reflects the functional capacity of the ovaries. 13 – 14 Changes in AMH levels after torsion-detorsion (T/D) can provide insights into the extent of ovarian reserve preservation. Therefore, serum AMH levels can be used as an indicator of ovarian reserve after the ovarian T/D. Asprosin is a recently discovered protein hormone secreted by white adipose tissue and encoded by the Fibrillin 1 (FBN1) gene. 15 It plays a critical role in glucose homeostasis and energy metabolism, particularly in the liver. 16 Studies suggest that asprosin has anti-inflammatory effects mediated by cytokines and may regulate chronic inflammation. 17 Tissue and serum asprosin levels have been associated with metabolic disorders such as insulin resistance, type 2 diabetes, and obesity. 17 – 19 Given its role in energy homeostasis and response to metabolic stress, asprosin may serve as a biomarker in conditions such as ischemia and reperfusion, where cellular energy balance is disrupted. 19 – 20 Therefore, changes in asprosin levels may be valuable for evaluating metabolic stress during ovarian T/D processes. In this context, our study aims to assess tissue and serum asprosin levels to better understand the metabolic effects of T/D. During ischemia and reperfusion, oxidative stress arises through mechanisms such as mitochondrial dysfunction, ATP depletion, intracellular calcium accumulation, and cytoskeletal damage, all of which contribute to increased production of reactive oxygen species (ROS). 21 This oxidative stress damages cell membranes, DNA, and proteins, compromising tissue integrity. Antioxidant agents play a crucial role in preventing such damage. 21 – 22 N-acetylcysteine (NAC), known for its potent antioxidant properties, enhances glutathione synthesis, supports cellular defense mechanisms, and reduces free radical-induced damage. 23 In addition to its antioxidant effects, NAC also exhibits antimutagenic and anti-inflammatory properties and has been used to mitigate ischemia-reperfusion (I/R) injury in various organs. 23 – 24 However, research on the effects of NAC in ovarian T/D models is still insufficient. This study aims to evaluate the protective effects of NAC on ovarian tissue in an experimentally induced rat model of ovarian T/D. In this context, AMH levels are measured to assess ovarian reserve, while tissue and serum asprosin levels, along with total oxidant status (TOS), are evaluated to determine the extent of inflammation and metabolic stress associated with reperfusion. The findings of this study are expected to clarify the effects of NAC on these parameters and support the development of pharmacological strategies to prevent tissue damage following ovarian torsion. Material-Method 1-Animal model: This randomized controlled experimental study was approved by the Experimental Animal Ethics Committee of Fırat University Faculty of Medicine (Number: 2019/02; meeting date: January 30, 2018) and conducted in accordance with ethical guidelines by the Department of Obstetrics and Gynecology at Fırat University. The experimental phase took place in April 2019 at the Experimental Animal Research Center of Fırat University Faculty of Medicine. The animals were obtained and housed by the Experimental Research Center of Fırat University. All rats were kept in specially designed cages under controlled conditions, with a room temperature maintained at 22–25°C, a 12-hour light (07:00–19:00) and 12-hour dark (19:00–07:00) cycle, and daily cleaning of cage floors. Standard rat chow and tap water were provided ad libitum using steel feeders and glass water bottles. Daily care, including grooming and general hygiene, was administered in accordance with the NIH Guide for the Care and Use of Laboratory Animals (Institute of Laboratory Animal Research, National Research Council, Washington, DC, USA). All investigators involved were certified in the use of experimental animals. Experimental protocol: Experimental protocol: Thirty-five female Wistar albino rats, aged 8 to 10 weeks, were randomly divided into five equal groups (n = 7 per group): Group I (Control), Group II (Sham), Group III (NAC), Group IV ( T/D), and Group V (T/D + NAC). For anesthesia, ketamine was administered intraperitoneally at a dose of 50 mg/kg to the subjects. Under anesthesia, the abdominal and cervical regions of the rats were shaved, and antisepsis was achieved using povidone-iodine. All animals subsequently underwent surgical intervention. A midline lower abdominal incision of approximately 2.5–3 cm was made, and the skin and subcutaneous tissues were dissected. In rats with normal-appearing intra-abdominal organs, the intestines were gently retracted laterally to the right to access the ovaries, which were identified by tracing the left uterine horn. Group I (Control group) To establish baseline values, a laparotomy was performed without any additional intervention. After visualization of the uterus and adnexa, the abdomen was closed. Three hours later, a second laparotomy was performed, during which bilateral ovaries were excised and blood samples were collected. The rats were then sacrificed. Group II (Sham group) In this group, the upper and lower poles of the ovaries were sutured. After 3 hours, the sutures were removed. At the 6th hour, bilateral ovaries and blood samples were collected, and the rats were sacrificed. Group III (NAC group) A single intraperitoneal dose of NAC (100 mg/kg) was administered 30 minutes prior to laparotomy. Following visualization of the uterus and adnexa, the abdomen was closed. After 3 hours, a second laparotomy was performed to collect bilateral ovaries and blood samples. The rats were then sacrificed. Group IV (T/D group) After laparotomy, the bilateral adnexa and ovaries were rotated 720 degrees clockwise and secured to the lateral abdominal wall with a 5/0 polydiaxone suture to prevent rotation. After 3 hours, a second laparotomy was performed, the fixation suture was removed, and detorsion was carried out. Based on literature, ischemia is known to develop in ovarian tissue after 3 hours of torsion. In our study, we applied a torsion period of 3 hours followed by 3 hours of detorsion. Three hours post-detorsion, bilateral ovaries were excised, blood samples were collected, and the rats were sacrificed. Group V (T/D + NAC group) Laparotomy was performed, and the bilateral adnexa and ovaries were rotated 720 degrees clockwise and secured to the lateral abdominal wall with a 5/0 polydiaxone suture. A single intraperitoneal dose of NAC (100 mg/kg) was administered 30 minutes prior to detorsion. Three hours after torsion, a second laparotomy was performed, the fixation suture was removed, and detorsion was carried out. Three hours post-detorsion, a final laparotomy was performed, bilateral ovaries were excised, and blood samples were collected. The rats were then sacrificed. Biochemical Study All biochemical analyses were conducted in the Medical Biochemistry Laboratory at Fırat University Faculty of Medicine. Blood samples were collected in biochemistry tubes containing aprotinin, then centrifuged at 3500 rpm for 5 minutes to separate the serum. The serum samples were stored at -80°C until the day of analysis. Serum TOS, Asprosin and AMH Levels Measurement TOS levels in serum samples were measured using the Enzyme Linked Immuno Sorbent Assay (ELISA) method. The Rat TOS ELISA kit (Catalog No: YLA1392Ra, YL Biotechnolog Co., Ltd, Shanghai, China) was used according to the manufacturer's instructions. The measurement range of the kit was 0.02-60 U/ml, with an intra-assay CV value < 10%, inter-assay CV value < 12%, and sensitivity of 0.013 U/ml. Plate washing was performed using a Bio-Tek ELX50 automatic washer (BioTek Instruments, USA), and absorbance readings were taken with a ChroMate Microplate Reader P4300 (Awareness Technology Instruments, USA). Results were reported in U/ml for serum samples. Asprosin levels in rat serum samples were measured using the ELISA method with the Rel Assay Rat Asprosin ELISA kit (REF: YX-011916R, LOT: 201806, Mega Tıp Sanayi ve Tic. Ltd, Gaziantep, Turkey) following the manufacturer's instructions. The intra-assay CV was < 10%, inter-assay CV was < 12%, and the sensitivity was 0.054 ng/mL. Plate washing was performed using a Bio-Tek ELX50 automatic washer (BioTek Instruments, USA), and absorbance readings were taken at 450 nm using a Bio-Tek ELX800 ELISA reader (BioTek Instruments, USA). The concentrations of prepared standards were entered into the Bio-Tek ELX800 and plotted on the standard curve. Results were calculated based on the standard curve and adjusted for the dilution factor, expressed in ng/mL. AMH levels in rat serum samples were quantified using the ELISA method with the AMH ELISA kit (Andygene Biotechnology Co., Ltd, Beijing, China), following the procedures outlined in the kit catalog. The assay sensitivity was 0.5 ng/mL. Absorbance readings were taken at 450 nm using a Multiskan™ FC Microplate Photometer ELISA reader (ThermoFisher). Preparation of tissue samples Ovarian tissues from each group were fixed in a 10% formaldehyde solution and washed with tap water. The tissues were subsequently subjected to a routine histological processing protocol (Table 1 ) and embedded in paraffin blocks. Sections of 4–6 µm thickness were obtained from the paraffin blocks and mounted onto polylysine-coated slides. Deparaffinized tissues were passed through a graded alcohol series and subjected to antigen retrieval by microwaving in citrate buffer solution (pH 6) at 750W for 15 minutes. Following the heating process, the tissues were allowed to cool at room temperature for approximately 25 minutes, washed with Phosphate Buffered Saline (PBS, P4417, Sigma-Aldrich, USA) for 3 × 5 minutes, and incubated with hydrogen peroxide block solution for 5 minutes to inhibit endogenous peroxidase activity (Hydrogen Peroxide Block, TA-125-HP, Lab Vision Corporation, USA). The tissues were then washed with PBS for 3 × 5 minutes and incubated with Ultra V Block (TA-125-UB, Lab Vision Corporation, USA) solution for 5 minutes to block non-specific staining. Subsequently, the primary antibody was diluted 1:200. Table 1 Histological Processing Series No. Procedure Duration 1 70% Alcohol 2 hours 2 80% Alcohol 1.5 hours 3 96% Alcohol I 30 minutes 4 96% Alcohol II 30 minutes 5 100% Alcohol I 30 minutes 6 100% Alcohol II 30 minutes 7 Alcohol + Xylene 15 minutes 8 Xylene I 15 minutes 9 Xylene II 15 minutes 10 Soft Paraffin + Xylene 45 minutes 11 Soft Paraffin 1 hour 12 Soft Paraffin + Hard Paraffin 1.5 hours 13 Hard Paraffin 3 hours 14 Inlaid The tissues were incubated with the primary antibody (anti-asprosine antibody, FNab09797, Fine Test, China) for 60 minutes in a humidified environment at room temperature. Following primary antibody incubation, the tissues were washed with PBS for 3 × 5 minutes and incubated with the secondary antibody (biotinylated Goat Anti-Polyvalent (anti-mouse/rabbit IgG), TP-125-BN, Lab Vision Corporation, USA) for 30 minutes at room temperature in a humidified environment. After secondary antibody application, the tissues were again washed with PBS for 3 × 5 minutes and incubated with Streptavidin Peroxidase (TS-125-HR, Lab Vision Corporation, USA) for 30 minutes in a humidified environment at room temperature, then placed in PBS. The tissues were subsequently incubated with 3-amino-9-ethylcarbazole (AEC) Substrate + AEC Chromogen (AEC Substrate, TA-015 and AEC Chromogen, TA-002-HAC, Lab Vision Corporation, USA) solution. After obtaining the image signal under a light microscope, the tissues were washed with PBS. The tissues were counterstained with Mayer's hematoxylin, washed with PBS and distilled water, and then mounted with the appropriate mounting solution (Large Volume Vision Mount, TA-125-UG, Lab Vision Corporation, USA). The slides were examined, evaluated, and photographed under a Leica DM500 microscope (Leica DFC295). A histoscore was determined based on the prevalence and severity of immunoreactivity. The prevalence was categorized as follows: 0.1 for < 25%, 0.4 for 26–50%, 0.6 for 51–75%, and 0.9 for 76–100%. The severity was scored as: 0 for none, + 0.5 for very little, + 1 for little, + 2 for moderate, and + 3 for severe. The histoscore was calculated by multiplying the prevalence and severity values. Statistical analysis Power analysis was conducted at 80% power and a 0.05 significance level for the variable with the largest standard deviation among the study variables. The analysis determined that each group should consist of at least 5 and at most 7 subjects. Statistical analysis was performed using SPSS 22.0 software (IBM Corp., Armonk, NY, USA). The Kruskal-Wallis test was employed for overall comparison between more than two groups, while the Mann-Whitney U test was used for pairwise comparisons. Data that were not normally distributed are presented as median (minimum-maximum). A p-value of less than 0.05 was considered statistically significant. Results Biochemical Findings In the biochemical analysis conducted to assess serum TOS levels across all groups, similar TOS levels were observed in Control, Sham, and NAC groups. Compared to the control group, T/D group showed a statistically significant increase in TOS levels (p = 0.016). However, TOS levels were significantly reduced in the T/D + NAC group when compared to T/D group (p = 0.032) (Table 2 ). Table 2 Serum TOS Levels Groups TOS Median (min-max) P P* Group 1 (Control) 2,28 (1,29 − 3,12) 0,001 1–2: 0,931 1–3: 0,063 1–4: 0,016 1–5: 0,008 2–3: 0,038 2–4: 0,010 2–5: 0,004 3–4: 0,029 3–5: 0,090 4–5: 0,032 Group 2 (Sham) 2,12 (1,35 − 2,95) Group 3 (NAC) 3,51 (2,49 − 4,66) Group 4 (Torsion) 12,16 (7,59 − 15,16) a Group 5 (Torsion + NAC) 5,38 (3,39 − 8,54) ab Values are presented as median (min–max). ᵃ: Compared to the Control group, ᵇ: Compared to the Torsion group (p < 0.05), P: Multiple comparison value (Kruskal-Wallis test), P*: Pairwise comparison value (Mann-Whitney U test). Asprosin levels were comparable between the control and sham groups. Compared to the control group, asprosin levels were significantly reduced in the NAC (p = 0.002), T/D (p = 0.003), and T/D + NAC (p = 0.010) groups (overall p = 0.016). Furthermore, when compared to the T/D group, asprosin levels were significantly higher in the T/D + NAC group (p = 0.008) (Table 3 ). Table 3 Serum Asprosin Levels Groups Serum Asprosin Levels Median (min-max) P P* Group 1 (Control) 1,39 (1,04 − 1,78) 0,001 1–2: 0,181 1–3: 0,002 1–4: 0,003 1–5: 0,010 2–3: 0,002 2–4: 0,004 2–5: 0,009 3–4: 0,149 3–5: 0,755 4–5: 0,008 Group 2 (Sham) 1,66 (1,21 − 2,21) Group 3 (NAC) 0,81 (0,16 − 1,26) a Group 4 (Torsion) 0,47 (0,36 − 0,61) a Group 5 (Torsion + NAC) 0,96 (0,74 − 1,22) ab Values are presented as median (min–max). ᵃ: Compared to the Control group, ᵇ: Compared to the Torsion group (p < 0.05), P: Multiple comparison value (Kruskal-Wallis test), P*: Pairwise comparison value (Mann-Whitney U test). AMH levels were similar in the control group and all other groups and no statistical difference was observed (Table 4 ). Table 4 Serum AMH Levels Groups Serum AMH Levels Median (min-max) P P* Group 1 (Control) 13,70 (11,94 − 15,07) 0,834 1–2: 0,628 1–3: 0,620 1–4: 0,456 1–5: 1,000 2–3: 1,000 2–4: 0,628 2–5: 0,534 3–4: 0,710 3–5: 0,805 4–5: 0,318 Group 2 (Sham) 14,29 (12,71 − 16,53) Group 3 (NAC) 14,16 (11,43 − 17,04) Group 4 (Torsion) 14,63 (11,44 − 16,97) Group 5 (Torsion + NAC) 13,84 (12,06–16,21) Values are presented as median (min–max). P: Multiple comparison value (Kruskal-Wallis test), P*: Pairwise comparison value (Mann-Whitney U test). Immunohistochemical Findings Asprosin Immunoreactivity Immunohistochemical staining for asprosin immunoreactivity under light microscopy revealed asprosin expression in the ovarian stroma (indicated by black stars). Asprosin immunoreactivity in ovarian tissue was similar among the Control (Fig. 1 ), Sham (Fig. 2 ), and NAC (Fig. 3 ) groups (Table 5 ). Compared to the control group, a significant decrease in asprosin immunoreactivity was observed in the T/D group (p = 0.002) (Table 5 , Fig. 4 ). However, when compared to the T/D group, asprosin immunoreactivity was significantly increased in the T/D + NAC group (p = 0.002) (Table 5 , Fig. 5 ). Table 5 Asprosin histoscore Groups Asprosin histoscore Median (min-max) P P* Group 1 (Control) 0,91 (0,80 − 1,20) 0,003 1–2: 0,699 1–3: 0,792 1–4: 0,002 1–5: 0,180 2–3: 0,931 2–4: 0,002 2–5: 0,310 3–4: 0,004 3–5: 0,329 4–5: 0,002 Group 2 (Sham) 0,83 (0,60 − 0,90) Group 3 (NAC) 0,88 (0,60 − 1,20) Group 4 (Torsion) 0,25 (0,20 − 0,45) a Group 5 (Torsion + NAC) 0,74 (0,45 − 0,90) b Values are presented as median (min–max). ᵃ: Compared to the Control group, ᵇ: Compared to the Torsion group (p < 0.05), P: Multiple comparison value (Kruskal-Wallis test), P*: Pairwise comparison value (Mann-Whitney U test). Discussion The findings of the current study offer valuable insights into the effects of NAC on ovarian reserve and asprosin levels in an ovarian T/D rat model. Ovarian torsion, a clinical condition leading to ischemia and reperfusion injury, poses a significant risk to ovarian health, and our study aimed to evaluate the potential protective role of NAC, an antioxidant known for its ability to combat oxidative stress. The increase in oxidative stress associated with ovarian torsion and subsequent detorsion is a well-established phenomenon, primarily driven by the overproduction of ROS, which cause cellular damage. Previous studies have demonstrated that ischemia-reperfusion injury in ovarian tissue is mediated through ROS generation, leading to damage to cell membranes, mitochondrial dysfunction, and follicular apoptosis. 25 – 26 NAC has been reported to act as a potent antioxidant by scavenging ROS, enhancing intracellular glutathione levels, and functioning as a reducing agent. 27 In a study on testicular torsion, a urological emergency, NAC was found to be effective in preventing reperfusion injury in the testis, though it did not prevent the reduction in spermatid count. 28 NAC has been shown to reduce oxidative stress and improve ovulation and pregnancy rates in infertile women with polycystic ovary syndrome (PCOS). 29 Ersoy et al. reported that NAC reduced oxidative damage and provided superior ovarian protection compared to enoxaparin sodium in a rat model of ovarian torsion. 30 Additionally, another study demonstrated that NAC may reduce oxidative damage and improve endothelial functions in diabetes patients . 31 Collectively, these findings suggest that the protective effects of NAC are primarily attributable to its ability to reduce oxidative stress and enhance tissue antioxidant capacity. One of the most significant findings in our study is the marked reduction in TOS in the T/D + NAC group (p = 0.032) compared to the T/D group. This significant decrease highlights the antioxidant potential of NAC in effectively neutralizing ROS and preserving ovarian tissue integrity and function. These results are in line with prior research suggesting that NAC-mediated enhancement of intracellular glutathione and reduction of oxidative stress may be beneficial in various models of ischemia-reperfusion injury. Asprosin, the C-terminal cleavage product of profibrillin, is a recently identified adipokine with a potent regulatory role in metabolism.³² In addition to stimulating hepatic gluconeogenesis, which elevates blood insulin and glucose levels, asprosin also directly promotes pancreatic β-cell apoptosis.³²–³³ It influences glucose homeostasis and energy metabolism. The role of asprosin—as a hormone linked to metabolic stress and energy regulation—emerged as another intriguing aspect of our study. In this context, considering that ovarian torsion and reperfusion affect energy metabolism, changes in asprosin levels in the T/D group were anticipated. Thus, we evaluated asprosin levels in reperfusion injury following ovarian torsion and detorsion, given its established role in energy metabolism and glucose homeostasis. Previous studies have demonstrated an association between altered asprosin levels and insulin resistance, diabetes, metabolic syndrome, obesity, and PCOS.³⁴–³⁶In our study, serum asprosin levels were significantly reduced in the T/D group (p = 0.003), reflecting metabolic disturbances and cellular stress resulting from ischemia. This reduction may indicate impaired glucose regulation and energy metabolism in ovarian tissue, as asprosin plays a critical role in maintaining blood glucose levels under stress conditions. Variations in asprosin levels observed between our study and previous studies involving different organs may be attributed to tissue-specific receptor expression and the influence of hormonal environments. 37 – 38 Furthermore, effects of asprosin on energy metabolism may vary depending on the tissue type. 39 – 40 Although there is a growing body of research on asprosin, further studies are needed to fully elucidate its functions and mechanisms of action. Some studies have suggested that NAC treatment may lead to elevated serum asprosin levels and has been linked to improvements in various disease states, particularly metabolic disorders. 41 – 42 Similarly, in our study, we found that serum asprosin levels (p = 0.008) and ovarian asprosin immunoreactivity (p = 0.002) were significantly higher in the T/D + NAC group compared to the T/D group. The increase in serum asprosin levels and asprosin immunoreactivity after NAC treatment further supports the idea that NAC not only reduces oxidative damage but may also help restore the metabolic equilibrium of the ovaries following ischemia-reperfusion injury. This result is in line with prior studies that have suggested asprosin's involvement in inflammatory processes and tissue repair, though its role in ovarian ischemia-reperfusion injury remains underexplored. Interestingly, asprosin levels were not significantly different between the control and sham groups, which further supports the specificity of the observed changes in the T/D and T/D + NAC groups. It is noteworthy that asprosin's role in the context of ovarian torsion and detorsion requires further investigation to fully understand its implications as a biomarker of metabolic stress. AMH is widely used as a biomarker for assessing ovarian reserve, as it reflects the number of antral follicles and ovarian functionality. 43 – 44 Many studies have been conducted on serum AMH levels indicating ovarian reserve. Low AMH levels have been shown to be associated with reproductive dysfunction, including diminished ovarian reserve and impaired ovarian function. 45 – 46 Thus, we measured serum AMH values ​​to evaluate ovarian reserve after the T/D and NAC treatment. We predicted that there would be changes in serum AMH levels in our study. In contrast to the changes observed in oxidative stress markers and asprosin levels, AMH levels did not show any significant differences among the groups. The lack of significant changes in AMH levels in this study could indicate that, despite the evident oxidative stress and changes in other metabolic markers, the degree of ovarian damage may not have been severe enough to influence AMH secretion. Alternatively, the duration of the ischemic event and the degree of injury in the T/D model might not have been sufficient to cause a measurable change in AMH levels, as these changes are often more pronounced in longer-term or more severe ovarian injuries. The results of this study suggest that NAC administration provides a protective effect against oxidative stress and may help to mitigate metabolic dysregulation in ovarian tissue following torsion-detorsion injury. By reducing oxidative damage and promoting recovery of asprosin levels, NAC may help preserve ovarian function, although further studies are needed to explore its long-term efficacy and potential clinical applications. The beneficial effects of NAC observed in this study support its potential as a therapeutic intervention for preventing ovarian damage in cases of ovarian torsion. Given that NAC is an already beneficial drug with a well-established safety profile, its use in clinical settings for ovarian torsion and other gynecological emergencies could be an accessible and cost-effective option to reduce tissue injury and preserve ovarian reserve. Limitations and Future Directions While this study provides valuable information on the effects of NAC in a rat model of ovarian torsion-detorsion, there are certain limitations. First, the study was conducted in an animal model, and while this provides valuable insights, further clinical studies are needed to translate these findings to human patients. Additionally, the mechanisms by which NAC influences asprosin expression and metabolic balance in ovarian tissues remain unclear and warrant further investigation. Finally, the duration of the torsion period and the use of NAC as a single dose may not fully replicate the clinical scenario, where delays in diagnosis and treatment are common. Therefore, future studies should explore different treatment regimens, timing of administration, and longer follow-up periods to better understand the clinical potential of NAC in ovarian torsion. Conclusion In conclusion, this study demonstrates that NAC effectively reduces oxidative stress and mitigates the adverse metabolic effects associated with ovarian torsion-detorsion injury. By partially restoring asprosin levels and reducing TOS, NAC holds promise as a therapeutic agent for protecting ovarian tissue in cases of ischemia-reperfusion injury. While AMH levels remained unchanged, the significant biochemical and histological improvements observed in the NAC-treated group suggest a potential role for NAC in preserving ovarian reserve following torsion-detorsion events. Future research should focus on optimizing NAC treatment protocols and investigating its long-term effects in clinical settings. Abbreviations The following abbreviations are used in this manuscript: T/D, torsion-detorsion; TOS, total oxidant status; ROS, reactive oxygen species; NAC, N-acetylcysteine; AMH, anti-Müllerian hormone; I/R, ischemia-reperfusion. Declarations Author Contributions Conceptualization, G.B., T.K. and A.A.; methodology, G.B, T.K and A.A.; software, C.S. and A.B.; validation, G.B; formal analysis, A.B. and C.S.; investigation, G.B., T.K. and A.A.; resources, G.B.; data curation, G.B. and A.A.; writing—original draft preparation, G.B.; writing—review and editing, G.B., A.B. and C.S.; visualization, A.B.; supervision, G.B and T.K.; project administration, G.B.; funding acquisition, G.B. All authors have read and agreed to the published version of the manuscript. Funding This research has not received any specific grants from funding organizations in the public, commercial or non-profit sectors. Informed Consent Statement Informed consent was obtained from all subjects involved in the study. Data Availability Statement Data are contained within the article. Conflicts of Interest The authors declare no conflicts of interest. References Bedekar K, McInnes A, Burgess W. Ovarian torsion: determining the presenting features and where the delays occur. N Z Med J 2025 Apr 11;138(1613):79-86. https://doi.org/10.26635/6965.6809 Huang C, Hong MK, Ding DC. A review of ovary torsion. Tzu Chi Med J. 2017 Jul-Sep;29(3):143-147. doi: 10.4103/tcmj.tcmj_55_17. https://doi.org/10.4103/tcmj.tcmj_55_17 Chang-Patel EJ, Palacios-Helgeson LK, Gould CH. Adnexal torsion: a review of diagnosis and management strategies. Curr Opin Obstet Gynecol. 2022 Aug 1;34(4):196-203. https://doi.org/10.1097/GCO.0000000000000787. Bridwell RE, Koyfman A, Long B. High risk and low prevalence diseases: Ovarian torsion. Am J Emerg Med. 2022 Jun; 56: 145-150. https://doi.org/10.1016/j.ajem.2022.03.046. Zhu TW, Li XL. Ovarian Torsion: A Review of the Evidence. 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Evaluation of the protective effects of gossypin for ischemia/reperfusion injury in ovary tissue. J Obstet Gynaecol Res. 2022 Mar;48(3):748-756. https://doi.org/10.1111/jog.15127. Osmanlıoğlu Ş, Arslan M, Dağ RO, Yığman Z, Ceyhan MŞ, Er F, Kavutçu M. Artemisinin reduces acute ovarian ischemia-reperfusion injury in rats. Reprod Toxicol. 2023 Aug; 119: 108417. https://doi.org/10.1016/j.reprotox.2023. Yilmaz F, Ilgen O, Mankan A, Yilmaz B, Kurt S. The effects of berberine on ischemia-reperfusion injuries in an experimental model of ovarian torsion. Clin Exp Reprod Med. 2023 Dec;50(4):292-298. https://doi.org/10.5653/cerm.2023. Moolhuijsen LME, Visser JA. Anti-Mullerian Hormone and Ovarian Reserve: Update on Assessing Ovarian Function. J Clin Endocrinol Metab. 2020 Nov 1;105(11):3361-73. https://doi.org/10.1210/clinem/dgaa513. Yildiz S, Moolhuijsen LME, Visser JA. The Role of Anti-Mullerian Hormone in Ovarian Function. Semin Reprod Med. 2024 Mar;42(1):15-24. https://doi.org/10.1055/s-0044-1786732. di Clemente N, Racine C, Pierre A, Taieb J. Anti-Mullerian Hormone in Female Reproduction. Endocr Rev. 2021 Nov 16;42(6):753-782. https://doi.org/10.1210/endrev/bnab012. Ovali, M. A., & Bozgeyik, I. (2022). Asprosin, a C-Terminal Cleavage Product of Fibrillin 1 Encoded by the FBN1 Gene, in Health and Disease. Molecular syndromology, 13(3), 175-183. https://doi.org/10.1159/000520333. Nedeva IS, et al. Circulating Asprosin Concentrations in Patients with Obesity and Carbohydrate Disturbances. Horm Metab Res. 2023 Apr;55(4):284-289. https://doi.org/10.1055/a-2033-6109. Mazur-Bialy AI. Asprosin Enhances Cytokine Production by a Co-Culture of Fully Differentiated Mature Adipocytes and Macrophages Leading to the Exacerbation of the Condition Typical of Obesity-Related Inflammation. Int J Mol Sci. 2023 Mar 17;24(6):5745. https://doi.org/10.3390/ijms24065745. Diao H, Li X, Xu Y, Xing X, Pang S. Asprosin, a novel glucogenic adipokine implicated in type 2 diabetes mellitus. J Diabetes Complications. 2023 Nov;37(11):108614. https://doi.org/10.1016/j.jdiacomp.2023. Zeng X, Sun X, He W, Xie J, Xin C. Relationship of asprosin and diabetes: a meta-analysis. BMC Endocr Disord. 2025 Jan 23;25(1):15. https://doi.org/10.1186/s12902-025-01843-1. Ozcan M, Ayar A. Endocrine Aspects of Pain Pathophysiology: Focus on Adipose Tissue. Neuroendocrinology. 2024;114(10):894-906. https://doi.org/10.1159/000539531. Kalyoncu Ş, et al. Melatonin attenuates ovarian ischemia reperfusion injury in rats by decreasing oxidative stress index and peroxynitrite. Turk J Med Sci. 2020 Oct 22;50(6):1513-1522. https://doi.org/10.3906/sag-2004-135. Çolak S, Koc K, Yıldırım S, Geyikoğlu F. Effects of boric acid on ovarian tissue damage caused by experimental ischemia/reperfusion. Biotech Histochem. 2022 Aug;97(6):415-422. https://doi.org/10.1080/10520295. Kumar P, et al. Supplementing Glycine and N-Acetylcysteine (GlyNAC) in Older Adults Improves Glutathione Deficiency, Oxidative Stress, Mitochondrial Dysfunction, Inflammation, Physical Function, and Aging Hallmarks: A Randomized Clinical Trial. J Gerontol A Biol Sci Med Sci. 2023 Jan 26;78(1):75-89. https://doi.org/10.1093/gerona/glac135. Raghu G, et al. The Multifaceted Therapeutic Role of N-Acetylcysteine (NAC) in Disorders Characterized by Oxidative Stress. Curr Neuropharmacol. 2021;19(8):1202-1224. https://doi.org/10.2174/1570159X19666201230144109. Barghi B, Shokoohi M, Khaki AA, Khaki A, Moghimian M, Soltani M. Eugenol improves tissue damage and oxidative stress in adult female rats after ovarian torsion/detorsion. J Obstet Gynaecol. 2021 Aug;41(6):933-938. https://doi.org/10.1080/01443615.2020. Demir M, et al. Metformin reduces ovarian ischemia reperfusion injury in rats by improving oxidative/nitrosative stress. aiwan J Obstet Gynecol. 2021 Jan;60(1):45-50. https://doi.org/10.1016/j.tjog.2020. Afzal S, Abdul Manap AS, Attiq A, Albokhadaim I, Kandeel M, Alhojaily SM. From imbalance to impairment: the central role of reactive oxygen species in oxidative stress-induced disorders and therapeutic exploration. Front Pharmacol. 2023; 14: 1269581. https://doi.org/10.3389/fphar.2023.1269581. Tangül, S. U., Çakmak, A. M., Çağlayan, O., & Bozdoğan, Ö. (2020). Prevention of the harmful effects of free oxygen radicals by using N-acetylcysteine in testicular torsion. Journal of Pediatric Urology, 16(1), 42-e1. https://doi.org/10.1016/j.jpurol.2019.10.028 Sandhu JK, et al. Oxidative Stress in Polycystic Ovarian Syndrome and the Effect of Antioxidant N-Acetylcysteine on Ovulation and Pregnancy Rate. Cureus. 2021 Sep 11;13(9): e17887. https://doi.org/10.7759/cureus.17887. Ersoy, G. S., Eken, M., Tal, R., Oztekin, D., Devranoglu, B., Kaygusuz, E. I., & Cevik, O. (2016). N-acetylcysteine leads to greater ovarian protection than enoxaparin sodium in a rat ovarian torsion model. Reproductive biomedicine online, 33(1), 93-101. https://doi.org/10.1016/j.rbmo.2016.03.009. Li X, Zou J, Lin A, Chi J, Hao H, Chen H, Liu Z. Oxidative Stress, Endothelial Dysfunction, and N-Acetylcysteine in Type 2 Diabetes Mellitus. Antioxid Redox Signal. 2024 Jun;40(16-18):968-989. https://doi.org/10.1089/ars.2023.0524. Diao H, Fan X, Li Z, Hou L, Dong Z, Pang S. Circulating asprosin concentrations in individuals with new-onset type 2 diabetes and prediabetes. Diabetes Res Clin Pract. 2024 Jul; 213:111730. https://doi.org/10.1016/j.diabres.2024. Wang C, et al. Asprosin aggravates nonalcoholic fatty liver disease via inflammation and lipid metabolic disturbance mediated by reactive oxygen species. Drug Dev Res. 2024 Jun;85(4):e22213. https://doi.org/10.1002/ddr.22213. Senyigit A, Durmus S, Gelisgen R, Uzun H. Oxidative Stress and Asprosin Levels in Type 2 Diabetic Patients with Good and Poor Glycemic Control. Biomolecules. 2024 Sep 5;14(9):1123. https://doi.org/10.3390/biom14091123. Hassan HJ, Hameed EK, Mohammad TU. Asprosin: the potential player in combined double diabetes and hypothyroidism. Ir J Med Sci. 2024 Dec;193(6):2915-2921. https://doi.org/10.1007/s11845-024-03758-7. Ulloque-Badaracco JR, et al. Asprosin levels in patients with type 2 diabetes mellitus, metabolic syndrome and obesity: A systematic review and meta-analysis. Diabetes Metab Syndr. 2024 Jul;18(7):103095. https://doi.org/10.1016/j.dsx.2024. Mishra I, et al. Protein tyrosine phosphatase receptor delta serves as the orexigenic asprosin receptor. Cell Metab. 2022 Apr 5;34(4):549-563.e8. https://doi.org/10.1016/j.cmet.2022.02.012. Corica D, et al. Asprosin serum levels and glucose homeostasis in children with obesity. Cytokine. 2021 Jun;142:155477. https://doi.org/10.1016/j.cyto.2021.155477. Hoffmann JG, Xie W, Chopra AR. Energy Regulation Mechanism and Therapeutic Potential of Asprosin. Diabetes. 2020 Apr;69(4):559-566. https://doi.org/10.2337/dbi19-0009. Hossein N, Mehdi M, Karim D, Mahdi M, Ghasemi E. Spirulina supplementation and circuit resistance training (CRT) reduce serum asprosin and appetite and improve energy balance in men with obesity and overweight. Hormones (Athens). 2025 Mar;24(1):23-31. https://doi.org/10.1007/s42000-024-00595-2. Fang YQ, et al. N-acetylcysteine supplementation improves endocrine-metabolism profiles and ovulation induction efficacy in polycystic ovary syndrome. J Ovarian Res. 2024 Oct 16;17(1):205. https://doi.org/10.1186/s13048-024-01528-8. Olesen HØ, et al. N-acetylcysteine protects ovarian follicles from ischemia-reperfusion injury in xenotransplanted human ovarian tissue. Hum Reprod. 2021 Jan 25;36(2):429-443. https://doi.org/10.1093/humrep/deaa291. Kacar E, et al. Asprosin-induced alterations in female rat puberty and reproductive hormonal profiles. Arch Physiol Biochem. 2025 Feb;131(1):24-32. https://doi.org/10.1080/13813455. Moolhuijsen LME, Visser JA. Anti-Mullerian Hormone and Ovarian Reserve: Update on Assessing Ovarian Function. J Clin Endocrinol Metab. 2020 Nov 1;105(11):3361-73. https://doi.org/10.1210/clinem/dgaa513. Cedars MI. Evaluation of Female Fertility-AMH and Ovarian Reserve Testing. J Clin Endocrinol Metab. 2022 May 17;107(6):1510-1519. https://doi.org/10.1210/clinem/dgac039. Harris BS, et al. Markers of ovarian reserve as predictors of future fertility. Fertil Steril. 2023 Jan;119(1):99-106. https://doi.org/10.1016/j.fertnstert.2022.10.014. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. 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20:23:09","currentVersionCode":1,"declarations":{"humanSubjects":false,"vertebrateSubjects":false,"conflictsOfInterestStatement":false,"humanSubjectEthicalGuidelines":false,"humanSubjectConsent":false,"humanSubjectClinicalTrial":false,"humanSubjectCaseReport":false,"vertebrateSubjectEthicalGuidelines":false},"doi":"10.21203/rs.3.rs-6623339/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6623339/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":82700504,"identity":"9e52da99-36d8-4e06-8548-762765f8522b","added_by":"auto","created_at":"2025-05-14 09:28:19","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":727230,"visible":true,"origin":"","legend":"\u003cp\u003eAsprosin immunoreactivity (indicated by black star) in the ovarian stroma of the control group.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-6623339/v1/7c25db3fe675c65c7d7e2baa.png"},{"id":82701162,"identity":"d66f4af5-82aa-4c1f-b1cf-49380b57b8c2","added_by":"auto","created_at":"2025-05-14 09:36:19","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":878861,"visible":true,"origin":"","legend":"\u003cp\u003eAsprosin immunoreactivity (indicated by black star) in the ovarian stroma of the sham group.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-6623339/v1/e3ba227ecb4894899c7dc27f.png"},{"id":82702291,"identity":"a2141f19-4814-43a1-9bb9-31ed78ca6869","added_by":"auto","created_at":"2025-05-14 09:44:19","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":803667,"visible":true,"origin":"","legend":"\u003cp\u003eAsprosin immunoreactivity (indicated by black star) in the ovarian stroma of the NAS group.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-6623339/v1/9dcde4009948cc972acb6d62.png"},{"id":82700509,"identity":"dfb8e8fa-5f98-4308-acf5-e37f77b42298","added_by":"auto","created_at":"2025-05-14 09:28:19","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":738944,"visible":true,"origin":"","legend":"\u003cp\u003eAsprosin immunoreactivity (indicated by black star) in the ovarian stroma of the torsion group.\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-6623339/v1/d8d2bc7b5bae49a57a0f4c42.png"},{"id":82700511,"identity":"cbac55fd-fd5c-4e8b-9347-f404d34eef0e","added_by":"auto","created_at":"2025-05-14 09:28:19","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":788962,"visible":true,"origin":"","legend":"\u003cp\u003eAsprosin immunoreactivity (indicated by black star) in the ovarian stroma of the torsion+NAS group.\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-6623339/v1/a1e137809bc4259a5a53848c.png"},{"id":96708055,"identity":"2e48934b-c85e-42e4-839d-c5b4e180dcc4","added_by":"auto","created_at":"2025-11-25 09:54:26","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":5639622,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6623339/v1/5048d2a1-7c46-410d-8d2d-47f90b7d56af.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"The Effects Of N-Acetylcysteine on Asprosine Immunoreactivity in A Rat Ovarian Torsion-Detorsion Model","fulltext":[{"header":"Introduction","content":"\u003cp\u003eOvarian torsion, defined as the complete or partial rotation of the ovary, fallopian tube, or both around their vascular axis, is a gynecological emergency with a prevalence of 2.7%.\u003csup\u003e1\u003c/sup\u003e It can affect women of all ages, and delayed diagnosis can negatively impact ovarian reserve.\u003csup\u003e\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e\u003c/sup\u003e Common causes include ovarian masses and ovulation induction.\u003csup\u003e\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u003c/sup\u003e However, the exact mechanisms leading to necrosis after torsion\u0026mdash;such as the degree and duration of rotation\u0026mdash;remain unclear.\u003csup\u003e\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u003c/sup\u003e This is because necrosis may not occur even in the presence of vascular pedicle twisting, whereas infarction and necrosis can sometimes develop due to venous congestion and edema despite preserved arterial blood flow.\u003csup\u003e\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u003c/sup\u003e Excluding prolonged ischemia, many torsion cases have shown preserved ovarian function.\u003csup\u003e\u003cspan additionalcitationids=\"CR7\" citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e\u003c/sup\u003e Therefore, early detection and prompt detorsion are the primary goals in the diagnosis and management of ovarian torsion.\u003csup\u003e\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e\u003c/sup\u003e However, the reperfusion process following detorsion can induce oxidative stress through the activation of free oxygen radicals and inflammatory cells.\u003csup\u003e\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e\u003c/sup\u003e Reperfusion is believed to be a major mechanism of tissue damage, independent of ischemia itself.\u003csup\u003e\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e\u003c/sup\u003e Although numerous studies have investigated ovarian torsion, the effects of reperfusion on ovarian reserve and energy metabolism remain insufficiently understood.\u003c/p\u003e \u003cp\u003eAnti-Mullerian Hormone (AMH) and antral follicle count are widely used parameters to assess ovarian reserve.\u003csup\u003e\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e\u003c/sup\u003e AMH is produced by growing follicles and reflects the functional capacity of the ovaries.\u003csup\u003e\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e\u003c/sup\u003e Changes in AMH levels after torsion-detorsion (T/D) can provide insights into the extent of ovarian reserve preservation. Therefore, serum AMH levels can be used as an indicator of ovarian reserve after the ovarian T/D.\u003c/p\u003e \u003cp\u003eAsprosin is a recently discovered protein hormone secreted by white adipose tissue and encoded by the Fibrillin 1 (FBN1) gene.\u003csup\u003e\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e\u003c/sup\u003e It plays a critical role in glucose homeostasis and energy metabolism, particularly in the liver.\u003csup\u003e\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e\u003c/sup\u003e Studies suggest that asprosin has anti-inflammatory effects mediated by cytokines and may regulate chronic inflammation.\u003csup\u003e\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e\u003c/sup\u003e Tissue and serum asprosin levels have been associated with metabolic disorders such as insulin resistance, type 2 diabetes, and obesity.\u003csup\u003e\u003cspan additionalcitationids=\"CR18\" citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e\u003c/sup\u003e Given its role in energy homeostasis and response to metabolic stress, asprosin may serve as a biomarker in conditions such as ischemia and reperfusion, where cellular energy balance is disrupted.\u003csup\u003e\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e\u003c/sup\u003e Therefore, changes in asprosin levels may be valuable for evaluating metabolic stress during ovarian T/D processes. In this context, our study aims to assess tissue and serum asprosin levels to better understand the metabolic effects of T/D.\u003c/p\u003e \u003cp\u003eDuring ischemia and reperfusion, oxidative stress arises through mechanisms such as mitochondrial dysfunction, ATP depletion, intracellular calcium accumulation, and cytoskeletal damage, all of which contribute to increased production of reactive oxygen species (ROS).\u003csup\u003e\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e\u003c/sup\u003e This oxidative stress damages cell membranes, DNA, and proteins, compromising tissue integrity. Antioxidant agents play a crucial role in preventing such damage.\u003csup\u003e\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e\u003c/sup\u003e N-acetylcysteine (NAC), known for its potent antioxidant properties, enhances glutathione synthesis, supports cellular defense mechanisms, and reduces free radical-induced damage.\u003csup\u003e\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e\u003c/sup\u003e In addition to its antioxidant effects, NAC also exhibits antimutagenic and anti-inflammatory properties and has been used to mitigate ischemia-reperfusion (I/R) injury in various organs.\u003csup\u003e\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e\u003c/sup\u003e However, research on the effects of NAC in ovarian T/D models is still insufficient.\u003c/p\u003e \u003cp\u003eThis study aims to evaluate the protective effects of NAC on ovarian tissue in an experimentally induced rat model of ovarian T/D. In this context, AMH levels are measured to assess ovarian reserve, while tissue and serum asprosin levels, along with total oxidant status (TOS), are evaluated to determine the extent of inflammation and metabolic stress associated with reperfusion. The findings of this study are expected to clarify the effects of NAC on these parameters and support the development of pharmacological strategies to prevent tissue damage following ovarian torsion.\u003c/p\u003e"},{"header":"Material-Method","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e1-Animal model:\u003c/h2\u003e \u003cp\u003eThis randomized controlled experimental study was approved by the Experimental Animal Ethics Committee of Fırat University Faculty of Medicine (Number: 2019/02; meeting date: January 30, 2018) and conducted in accordance with ethical guidelines by the Department of Obstetrics and Gynecology at Fırat University. The experimental phase took place in April 2019 at the Experimental Animal Research Center of Fırat University Faculty of Medicine.\u003c/p\u003e \u003cp\u003eThe animals were obtained and housed by the Experimental Research Center of Fırat University. All rats were kept in specially designed cages under controlled conditions, with a room temperature maintained at 22\u0026ndash;25\u0026deg;C, a 12-hour light (07:00\u0026ndash;19:00) and 12-hour dark (19:00\u0026ndash;07:00) cycle, and daily cleaning of cage floors. Standard rat chow and tap water were provided ad libitum using steel feeders and glass water bottles. Daily care, including grooming and general hygiene, was administered in accordance with the NIH Guide for the Care and Use of Laboratory Animals (Institute of Laboratory Animal Research, National Research Council, Washington, DC, USA). All investigators involved were certified in the use of experimental animals.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eExperimental protocol:\u003c/h3\u003e\n\u003cdiv class=\"Heading\"\u003eExperimental protocol:\u003c/div\u003e \u003cp\u003eThirty-five female Wistar albino rats, aged 8 to 10 weeks, were randomly divided into five equal groups (n\u0026thinsp;=\u0026thinsp;7 per group): Group I (Control), Group II (Sham), Group III (NAC), Group IV ( T/D), and Group V (T/D\u0026thinsp;+\u0026thinsp;NAC). For anesthesia, ketamine was administered intraperitoneally at a dose of 50 mg/kg to the subjects. Under anesthesia, the abdominal and cervical regions of the rats were shaved, and antisepsis was achieved using povidone-iodine. All animals subsequently underwent surgical intervention. A midline lower abdominal incision of approximately 2.5\u0026ndash;3 cm was made, and the skin and subcutaneous tissues were dissected. In rats with normal-appearing intra-abdominal organs, the intestines were gently retracted laterally to the right to access the ovaries, which were identified by tracing the left uterine horn.\u003c/p\u003e \u003cp\u003e \u003cstrong\u003eGroup I (Control group)\u003c/strong\u003e \u003cp\u003eTo establish baseline values, a laparotomy was performed without any additional intervention. After visualization of the uterus and adnexa, the abdomen was closed. Three hours later, a second laparotomy was performed, during which bilateral ovaries were excised and blood samples were collected. The rats were then sacrificed.\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cstrong\u003eGroup II (Sham group)\u003c/strong\u003e \u003cp\u003eIn this group, the upper and lower poles of the ovaries were sutured. After 3 hours, the sutures were removed. At the 6th hour, bilateral ovaries and blood samples were collected, and the rats were sacrificed.\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cstrong\u003eGroup III (NAC group)\u003c/strong\u003e \u003cp\u003eA single intraperitoneal dose of NAC (100 mg/kg) was administered 30 minutes prior to laparotomy. Following visualization of the uterus and adnexa, the abdomen was closed. After 3 hours, a second laparotomy was performed to collect bilateral ovaries and blood samples. The rats were then sacrificed.\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cstrong\u003eGroup IV (T/D group)\u003c/strong\u003e \u003cp\u003eAfter laparotomy, the bilateral adnexa and ovaries were rotated 720 degrees clockwise and secured to the lateral abdominal wall with a 5/0 polydiaxone suture to prevent rotation. After 3 hours, a second laparotomy was performed, the fixation suture was removed, and detorsion was carried out. Based on literature, ischemia is known to develop in ovarian tissue after 3 hours of torsion. In our study, we applied a torsion period of 3 hours followed by 3 hours of detorsion. Three hours post-detorsion, bilateral ovaries were excised, blood samples were collected, and the rats were sacrificed.\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cstrong\u003eGroup V (T/D\u0026thinsp;+\u0026thinsp;NAC group)\u003c/strong\u003e \u003cp\u003eLaparotomy was performed, and the bilateral adnexa and ovaries were rotated 720 degrees clockwise and secured to the lateral abdominal wall with a 5/0 polydiaxone suture. A single intraperitoneal dose of NAC (100 mg/kg) was administered 30 minutes prior to detorsion. Three hours after torsion, a second laparotomy was performed, the fixation suture was removed, and detorsion was carried out. Three hours post-detorsion, a final laparotomy was performed, bilateral ovaries were excised, and blood samples were collected. The rats were then sacrificed.\u003c/p\u003e \u003c/p\u003e\n\u003ch3\u003eBiochemical Study\u003c/h3\u003e\n\u003cp\u003eAll biochemical analyses were conducted in the Medical Biochemistry Laboratory at Fırat University Faculty of Medicine. Blood samples were collected in biochemistry tubes containing aprotinin, then centrifuged at 3500 rpm for 5 minutes to separate the serum. The serum samples were stored at -80\u0026deg;C until the day of analysis.\u003c/p\u003e\n\u003ch3\u003eSerum TOS, Asprosin and AMH Levels Measurement\u003c/h3\u003e\n\u003cp\u003eTOS levels in serum samples were measured using the Enzyme Linked Immuno Sorbent Assay (ELISA) method. The Rat TOS ELISA kit (Catalog No: YLA1392Ra, YL Biotechnolog Co., Ltd, Shanghai, China) was used according to the manufacturer's instructions. The measurement range of the kit was 0.02-60 U/ml, with an intra-assay CV value\u0026thinsp;\u0026lt;\u0026thinsp;10%, inter-assay CV value\u0026thinsp;\u0026lt;\u0026thinsp;12%, and sensitivity of 0.013 U/ml. Plate washing was performed using a Bio-Tek ELX50 automatic washer (BioTek Instruments, USA), and absorbance readings were taken with a ChroMate Microplate Reader P4300 (Awareness Technology Instruments, USA). Results were reported in U/ml for serum samples.\u003c/p\u003e \u003cp\u003eAsprosin levels in rat serum samples were measured using the ELISA method with the Rel Assay Rat Asprosin ELISA kit (REF: YX-011916R, LOT: 201806, Mega Tıp Sanayi ve Tic. Ltd, Gaziantep, Turkey) following the manufacturer's instructions. The intra-assay CV was \u0026lt;\u0026thinsp;10%, inter-assay CV was \u0026lt;\u0026thinsp;12%, and the sensitivity was 0.054 ng/mL. Plate washing was performed using a Bio-Tek ELX50 automatic washer (BioTek Instruments, USA), and absorbance readings were taken at 450 nm using a Bio-Tek ELX800 ELISA reader (BioTek Instruments, USA). The concentrations of prepared standards were entered into the Bio-Tek ELX800 and plotted on the standard curve. Results were calculated based on the standard curve and adjusted for the dilution factor, expressed in ng/mL.\u003c/p\u003e \u003cp\u003eAMH levels in rat serum samples were quantified using the ELISA method with the AMH ELISA kit (Andygene Biotechnology Co., Ltd, Beijing, China), following the procedures outlined in the kit catalog. The assay sensitivity was 0.5 ng/mL. Absorbance readings were taken at 450 nm using a Multiskan\u0026trade; FC Microplate Photometer ELISA reader (ThermoFisher).\u003c/p\u003e\n\u003ch3\u003ePreparation of tissue samples\u003c/h3\u003e\n\u003cp\u003eOvarian tissues from each group were fixed in a 10% formaldehyde solution and washed with tap water. The tissues were subsequently subjected to a routine histological processing protocol (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e) and embedded in paraffin blocks. Sections of 4\u0026ndash;6 \u0026micro;m thickness were obtained from the paraffin blocks and mounted onto polylysine-coated slides. Deparaffinized tissues were passed through a graded alcohol series and subjected to antigen retrieval by microwaving in citrate buffer solution (pH 6) at 750W for 15 minutes. Following the heating process, the tissues were allowed to cool at room temperature for approximately 25 minutes, washed with Phosphate Buffered Saline (PBS, P4417, Sigma-Aldrich, USA) for 3 \u0026times; 5 minutes, and incubated with hydrogen peroxide block solution for 5 minutes to inhibit endogenous peroxidase activity (Hydrogen Peroxide Block, TA-125-HP, Lab Vision Corporation, USA). The tissues were then washed with PBS for 3 \u0026times; 5 minutes and incubated with Ultra V Block (TA-125-UB, Lab Vision Corporation, USA) solution for 5 minutes to block non-specific staining. Subsequently, the primary antibody was diluted 1:200.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eHistological Processing Series\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"3\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNo.\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eProcedure\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eDuration\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e70% Alcohol\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2 hours\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e80% Alcohol\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.5 hours\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e96% Alcohol I\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e30 minutes\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e96% Alcohol II\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e30 minutes\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e100% Alcohol I\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e30 minutes\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e100% Alcohol II\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e30 minutes\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eAlcohol\u0026thinsp;+\u0026thinsp;Xylene\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e15 minutes\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eXylene I\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e15 minutes\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eXylene II\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e15 minutes\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSoft Paraffin\u0026thinsp;+\u0026thinsp;Xylene\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e45 minutes\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSoft Paraffin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1 hour\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSoft Paraffin\u0026thinsp;+\u0026thinsp;Hard Paraffin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.5 hours\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eHard Paraffin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3 hours\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eInlaid\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe tissues were incubated with the primary antibody (anti-asprosine antibody, FNab09797, Fine Test, China) for 60 minutes in a humidified environment at room temperature. Following primary antibody incubation, the tissues were washed with PBS for 3 \u0026times; 5 minutes and incubated with the secondary antibody (biotinylated Goat Anti-Polyvalent (anti-mouse/rabbit IgG), TP-125-BN, Lab Vision Corporation, USA) for 30 minutes at room temperature in a humidified environment. After secondary antibody application, the tissues were again washed with PBS for 3 \u0026times; 5 minutes and incubated with Streptavidin Peroxidase (TS-125-HR, Lab Vision Corporation, USA) for 30 minutes in a humidified environment at room temperature, then placed in PBS. The tissues were subsequently incubated with 3-amino-9-ethylcarbazole (AEC) Substrate\u0026thinsp;+\u0026thinsp;AEC Chromogen (AEC Substrate, TA-015 and AEC Chromogen, TA-002-HAC, Lab Vision Corporation, USA) solution. After obtaining the image signal under a light microscope, the tissues were washed with PBS. The tissues were counterstained with Mayer's hematoxylin, washed with PBS and distilled water, and then mounted with the appropriate mounting solution (Large Volume Vision Mount, TA-125-UG, Lab Vision Corporation, USA). The slides were examined, evaluated, and photographed under a Leica DM500 microscope (Leica DFC295).\u003c/p\u003e \u003cp\u003eA histoscore was determined based on the prevalence and severity of immunoreactivity. The prevalence was categorized as follows: 0.1 for \u0026lt;\u0026thinsp;25%, 0.4 for 26\u0026ndash;50%, 0.6 for 51\u0026ndash;75%, and 0.9 for 76\u0026ndash;100%. The severity was scored as: 0 for none, +\u0026thinsp;0.5 for very little, +\u0026thinsp;1 for little, +\u0026thinsp;2 for moderate, and +\u0026thinsp;3 for severe. The histoscore was calculated by multiplying the prevalence and severity values.\u003c/p\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eStatistical analysis\u003c/h2\u003e \u003cp\u003ePower analysis was conducted at 80% power and a 0.05 significance level for the variable with the largest standard deviation among the study variables. The analysis determined that each group should consist of at least 5 and at most 7 subjects. Statistical analysis was performed using SPSS 22.0 software (IBM Corp., Armonk, NY, USA). The Kruskal-Wallis test was employed for overall comparison between more than two groups, while the Mann-Whitney U test was used for pairwise comparisons. Data that were not normally distributed are presented as median (minimum-maximum). A p-value of less than 0.05 was considered statistically significant.\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003eBiochemical Findings\u003c/h2\u003e \u003cp\u003eIn the biochemical analysis conducted to assess serum TOS levels across all groups, similar TOS levels were observed in Control, Sham, and NAC groups. Compared to the control group, T/D group showed a statistically significant increase in TOS levels (p\u0026thinsp;=\u0026thinsp;0.016). However, TOS levels were significantly reduced in the T/D\u0026thinsp;+\u0026thinsp;NAC group when compared to T/D group (p\u0026thinsp;=\u0026thinsp;0.032) (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eSerum TOS Levels\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroups\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTOS\u003c/p\u003e \u003cp\u003eMedian (min-max)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eP\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eP*\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroup 1 (Control)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2,28 (1,29\u0026thinsp;\u0026minus;\u0026thinsp;3,12)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e0,001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e1\u0026ndash;2: 0,931\u003c/p\u003e \u003cp\u003e1\u0026ndash;3: 0,063\u003c/p\u003e \u003cp\u003e1\u0026ndash;4: 0,016\u003c/p\u003e \u003cp\u003e1\u0026ndash;5: 0,008\u003c/p\u003e \u003cp\u003e2\u0026ndash;3: 0,038\u003c/p\u003e \u003cp\u003e2\u0026ndash;4: 0,010\u003c/p\u003e \u003cp\u003e2\u0026ndash;5: 0,004\u003c/p\u003e \u003cp\u003e3\u0026ndash;4: 0,029\u003c/p\u003e \u003cp\u003e3\u0026ndash;5: 0,090\u003c/p\u003e \u003cp\u003e4\u0026ndash;5: 0,032\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroup 2 (Sham)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2,12 (1,35\u0026thinsp;\u0026minus;\u0026thinsp;2,95)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroup 3 (NAC)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3,51 (2,49\u0026thinsp;\u0026minus;\u0026thinsp;4,66)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroup 4 (Torsion)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e12,16 (7,59\u0026thinsp;\u0026minus;\u0026thinsp;15,16)\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroup 5 (Torsion\u0026thinsp;+\u0026thinsp;NAC)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e5,38 (3,39\u0026thinsp;\u0026minus;\u0026thinsp;8,54)\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"4\"\u003eValues are presented as median (min\u0026ndash;max).\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"4\"\u003eᵃ: Compared to the Control group,\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"4\"\u003eᵇ: Compared to the Torsion group (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05),\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"4\"\u003eP: Multiple comparison value (Kruskal-Wallis test),\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"4\"\u003eP*: Pairwise comparison value (Mann-Whitney U test).\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eAsprosin levels were comparable between the control and sham groups. Compared to the control group, asprosin levels were significantly reduced in the NAC (p\u0026thinsp;=\u0026thinsp;0.002), T/D (p\u0026thinsp;=\u0026thinsp;0.003), and T/D\u0026thinsp;+\u0026thinsp;NAC (p\u0026thinsp;=\u0026thinsp;0.010) groups (overall p\u0026thinsp;=\u0026thinsp;0.016). Furthermore, when compared to the T/D group, asprosin levels were significantly higher in the T/D\u0026thinsp;+\u0026thinsp;NAC group (p\u0026thinsp;=\u0026thinsp;0.008) (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eSerum Asprosin Levels\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroups\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSerum Asprosin Levels\u003c/p\u003e \u003cp\u003eMedian (min-max)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eP\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eP*\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroup 1 (Control)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1,39 (1,04\u0026thinsp;\u0026minus;\u0026thinsp;1,78)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e0,001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e1\u0026ndash;2: 0,181\u003c/p\u003e \u003cp\u003e1\u0026ndash;3: 0,002\u003c/p\u003e \u003cp\u003e1\u0026ndash;4: 0,003\u003c/p\u003e \u003cp\u003e1\u0026ndash;5: 0,010\u003c/p\u003e \u003cp\u003e2\u0026ndash;3: 0,002\u003c/p\u003e \u003cp\u003e2\u0026ndash;4: 0,004\u003c/p\u003e \u003cp\u003e2\u0026ndash;5: 0,009\u003c/p\u003e \u003cp\u003e3\u0026ndash;4: 0,149\u003c/p\u003e \u003cp\u003e3\u0026ndash;5: 0,755\u003c/p\u003e \u003cp\u003e4\u0026ndash;5: 0,008\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroup 2 (Sham)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1,66 (1,21\u0026thinsp;\u0026minus;\u0026thinsp;2,21)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroup 3 (NAC)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0,81 (0,16\u0026thinsp;\u0026minus;\u0026thinsp;1,26)\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroup 4 (Torsion)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0,47 (0,36\u0026thinsp;\u0026minus;\u0026thinsp;0,61)\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroup 5 (Torsion\u0026thinsp;+\u0026thinsp;NAC)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0,96 (0,74\u0026thinsp;\u0026minus;\u0026thinsp;1,22)\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"4\"\u003eValues are presented as median (min\u0026ndash;max).\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"4\"\u003eᵃ: Compared to the Control group,\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"4\"\u003eᵇ: Compared to the Torsion group (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05),\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"4\"\u003eP: Multiple comparison value (Kruskal-Wallis test),\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"4\"\u003eP*: Pairwise comparison value (Mann-Whitney U test).\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eAMH levels were similar in the control group and all other groups and no statistical difference was observed (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eSerum AMH Levels\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroups\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSerum AMH Levels\u003c/p\u003e \u003cp\u003eMedian (min-max)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eP\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eP*\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroup 1 (Control)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e13,70 (11,94\u0026thinsp;\u0026minus;\u0026thinsp;15,07)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e0,834\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e1\u0026ndash;2: 0,628\u003c/p\u003e \u003cp\u003e1\u0026ndash;3: 0,620\u003c/p\u003e \u003cp\u003e1\u0026ndash;4: 0,456\u003c/p\u003e \u003cp\u003e1\u0026ndash;5: 1,000\u003c/p\u003e \u003cp\u003e2\u0026ndash;3: 1,000\u003c/p\u003e \u003cp\u003e2\u0026ndash;4: 0,628\u003c/p\u003e \u003cp\u003e2\u0026ndash;5: 0,534\u003c/p\u003e \u003cp\u003e3\u0026ndash;4: 0,710\u003c/p\u003e \u003cp\u003e3\u0026ndash;5: 0,805\u003c/p\u003e \u003cp\u003e4\u0026ndash;5: 0,318\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroup 2 (Sham)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e14,29 (12,71\u0026thinsp;\u0026minus;\u0026thinsp;16,53)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroup 3 (NAC)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e14,16 (11,43\u0026thinsp;\u0026minus;\u0026thinsp;17,04)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroup 4 (Torsion)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e14,63 (11,44\u0026thinsp;\u0026minus;\u0026thinsp;16,97)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroup 5 (Torsion\u0026thinsp;+\u0026thinsp;NAC)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e13,84 (12,06\u0026ndash;16,21)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"4\"\u003eValues are presented as median (min\u0026ndash;max).\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"4\"\u003eP: Multiple comparison value (Kruskal-Wallis test),\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"4\"\u003eP*: Pairwise comparison value (Mann-Whitney U test).\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eImmunohistochemical Findings\u003c/h2\u003e \u003cdiv id=\"Sec12\" class=\"Section3\"\u003e \u003ch2\u003eAsprosin Immunoreactivity\u003c/h2\u003e \u003cp\u003eImmunohistochemical staining for asprosin immunoreactivity under light microscopy revealed asprosin expression in the ovarian stroma (indicated by black stars). Asprosin immunoreactivity in ovarian tissue was similar among the Control (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e), Sham (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e), and NAC (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e) groups (Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e). Compared to the control group, a significant decrease in asprosin immunoreactivity was observed in the T/D group (p\u0026thinsp;=\u0026thinsp;0.002) (Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e, Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). However, when compared to the T/D group, asprosin immunoreactivity was significantly increased in the T/D\u0026thinsp;+\u0026thinsp;NAC group (p\u0026thinsp;=\u0026thinsp;0.002) (Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e, Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab5\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eAsprosin histoscore\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroups\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eAsprosin histoscore Median (min-max)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eP\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eP*\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroup 1 (Control)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0,91 (0,80\u0026thinsp;\u0026minus;\u0026thinsp;1,20)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e0,003\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e1\u0026ndash;2: 0,699\u003c/p\u003e \u003cp\u003e1\u0026ndash;3: 0,792\u003c/p\u003e \u003cp\u003e1\u0026ndash;4: 0,002\u003c/p\u003e \u003cp\u003e1\u0026ndash;5: 0,180\u003c/p\u003e \u003cp\u003e2\u0026ndash;3: 0,931\u003c/p\u003e \u003cp\u003e2\u0026ndash;4: 0,002\u003c/p\u003e \u003cp\u003e2\u0026ndash;5: 0,310\u003c/p\u003e \u003cp\u003e3\u0026ndash;4: 0,004\u003c/p\u003e \u003cp\u003e3\u0026ndash;5: 0,329\u003c/p\u003e \u003cp\u003e4\u0026ndash;5: 0,002\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroup 2 (Sham)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0,83 (0,60\u0026thinsp;\u0026minus;\u0026thinsp;0,90)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroup 3 (NAC)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0,88 (0,60\u0026thinsp;\u0026minus;\u0026thinsp;1,20)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroup 4 (Torsion)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0,25 (0,20\u0026thinsp;\u0026minus;\u0026thinsp;0,45)\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroup 5 (Torsion\u0026thinsp;+\u0026thinsp;NAC)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0,74 (0,45\u0026thinsp;\u0026minus;\u0026thinsp;0,90)\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"4\"\u003eValues are presented as median (min\u0026ndash;max).\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"4\"\u003eᵃ: Compared to the Control group,\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"4\"\u003eᵇ: Compared to the Torsion group (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05),\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"4\"\u003eP: Multiple comparison value (Kruskal-Wallis test),\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"4\"\u003eP*: Pairwise comparison value (Mann-Whitney U test).\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eThe findings of the current study offer valuable insights into the effects of NAC on ovarian reserve and asprosin levels in an ovarian T/D rat model. Ovarian torsion, a clinical condition leading to ischemia and reperfusion injury, poses a significant risk to ovarian health, and our study aimed to evaluate the potential protective role of NAC, an antioxidant known for its ability to combat oxidative stress.\u003c/p\u003e \u003cp\u003eThe increase in oxidative stress associated with ovarian torsion and subsequent detorsion is a well-established phenomenon, primarily driven by the overproduction of ROS, which cause cellular damage. Previous studies have demonstrated that ischemia-reperfusion injury in ovarian tissue is mediated through ROS generation, leading to damage to cell membranes, mitochondrial dysfunction, and follicular apoptosis.\u003csup\u003e\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e\u003c/sup\u003e NAC has been reported to act as a potent antioxidant by scavenging ROS, enhancing intracellular glutathione levels, and functioning as a reducing agent.\u003csup\u003e\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e\u003c/sup\u003e In a study on testicular torsion, a urological emergency, NAC was found to be effective in preventing reperfusion injury in the testis, though it did not prevent the reduction in spermatid count.\u003csup\u003e\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e\u003c/sup\u003e NAC has been shown to reduce oxidative stress and improve ovulation and pregnancy rates in infertile women with polycystic ovary syndrome (PCOS).\u003csup\u003e\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e\u003c/sup\u003e Ersoy et al. reported that NAC reduced oxidative damage and provided superior ovarian protection compared to enoxaparin sodium in a rat model of ovarian torsion.\u003csup\u003e\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e\u003c/sup\u003e Additionally, another study demonstrated that NAC may reduce oxidative damage and improve endothelial functions in diabetes patients .\u003csup\u003e\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e\u003c/sup\u003e Collectively, these findings suggest that the protective effects of NAC are primarily attributable to its ability to reduce oxidative stress and enhance tissue antioxidant capacity. One of the most significant findings in our study is the marked reduction in TOS in the T/D\u0026thinsp;+\u0026thinsp;NAC group (p\u0026thinsp;=\u0026thinsp;0.032) compared to the T/D group. This significant decrease highlights the antioxidant potential of NAC in effectively neutralizing ROS and preserving ovarian tissue integrity and function. These results are in line with prior research suggesting that NAC-mediated enhancement of intracellular glutathione and reduction of oxidative stress may be beneficial in various models of ischemia-reperfusion injury.\u003c/p\u003e \u003cp\u003eAsprosin, the C-terminal cleavage product of profibrillin, is a recently identified adipokine with a potent regulatory role in metabolism.\u0026sup3;\u0026sup2; In addition to stimulating hepatic gluconeogenesis, which elevates blood insulin and glucose levels, asprosin also directly promotes pancreatic β-cell apoptosis.\u0026sup3;\u0026sup2;\u0026ndash;\u0026sup3;\u0026sup3; It influences glucose homeostasis and energy metabolism. The role of asprosin\u0026mdash;as a hormone linked to metabolic stress and energy regulation\u0026mdash;emerged as another intriguing aspect of our study. In this context, considering that ovarian torsion and reperfusion affect energy metabolism, changes in asprosin levels in the T/D group were anticipated. Thus, we evaluated asprosin levels in reperfusion injury following ovarian torsion and detorsion, given its established role in energy metabolism and glucose homeostasis. Previous studies have demonstrated an association between altered asprosin levels and insulin resistance, diabetes, metabolic syndrome, obesity, and PCOS.\u0026sup3;⁴\u0026ndash;\u0026sup3;⁶In our study, serum asprosin levels were significantly reduced in the T/D group (p\u0026thinsp;=\u0026thinsp;0.003), reflecting metabolic disturbances and cellular stress resulting from ischemia. This reduction may indicate impaired glucose regulation and energy metabolism in ovarian tissue, as asprosin plays a critical role in maintaining blood glucose levels under stress conditions. Variations in asprosin levels observed between our study and previous studies involving different organs may be attributed to tissue-specific receptor expression and the influence of hormonal environments.\u003csup\u003e\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e\u003c/sup\u003e Furthermore, effects of asprosin on energy metabolism may vary depending on the tissue type.\u003csup\u003e\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e\u003c/sup\u003e Although there is a growing body of research on asprosin, further studies are needed to fully elucidate its functions and mechanisms of action.\u003c/p\u003e \u003cp\u003eSome studies have suggested that NAC treatment may lead to elevated serum asprosin levels and has been linked to improvements in various disease states, particularly metabolic disorders.\u003csup\u003e\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e\u003c/sup\u003e Similarly, in our study, we found that serum asprosin levels (p\u0026thinsp;=\u0026thinsp;0.008) and ovarian asprosin immunoreactivity (p\u0026thinsp;=\u0026thinsp;0.002) were significantly higher in the T/D\u0026thinsp;+\u0026thinsp;NAC group compared to the T/D group. The increase in serum asprosin levels and asprosin immunoreactivity after NAC treatment further supports the idea that NAC not only reduces oxidative damage but may also help restore the metabolic equilibrium of the ovaries following ischemia-reperfusion injury. This result is in line with prior studies that have suggested asprosin's involvement in inflammatory processes and tissue repair, though its role in ovarian ischemia-reperfusion injury remains underexplored.\u003c/p\u003e \u003cp\u003eInterestingly, asprosin levels were not significantly different between the control and sham groups, which further supports the specificity of the observed changes in the T/D and T/D\u0026thinsp;+\u0026thinsp;NAC groups. It is noteworthy that asprosin's role in the context of ovarian torsion and detorsion requires further investigation to fully understand its implications as a biomarker of metabolic stress.\u003c/p\u003e \u003cp\u003eAMH is widely used as a biomarker for assessing ovarian reserve, as it reflects the number of antral follicles and ovarian functionality.\u003csup\u003e\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e\u003c/sup\u003e Many studies have been conducted on serum AMH levels indicating ovarian reserve. Low AMH levels have been shown to be associated with reproductive dysfunction, including diminished ovarian reserve and impaired ovarian function. \u003csup\u003e\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e\u003c/sup\u003e Thus, we measured serum AMH values ​​to evaluate ovarian reserve after the T/D and NAC treatment. We predicted that there would be changes in serum AMH levels in our study. In contrast to the changes observed in oxidative stress markers and asprosin levels, AMH levels did not show any significant differences among the groups. The lack of significant changes in AMH levels in this study could indicate that, despite the evident oxidative stress and changes in other metabolic markers, the degree of ovarian damage may not have been severe enough to influence AMH secretion. Alternatively, the duration of the ischemic event and the degree of injury in the T/D model might not have been sufficient to cause a measurable change in AMH levels, as these changes are often more pronounced in longer-term or more severe ovarian injuries.\u003c/p\u003e \u003cp\u003eThe results of this study suggest that NAC administration provides a protective effect against oxidative stress and may help to mitigate metabolic dysregulation in ovarian tissue following torsion-detorsion injury. By reducing oxidative damage and promoting recovery of asprosin levels, NAC may help preserve ovarian function, although further studies are needed to explore its long-term efficacy and potential clinical applications.\u003c/p\u003e \u003cp\u003eThe beneficial effects of NAC observed in this study support its potential as a therapeutic intervention for preventing ovarian damage in cases of ovarian torsion. Given that NAC is an already beneficial drug with a well-established safety profile, its use in clinical settings for ovarian torsion and other gynecological emergencies could be an accessible and cost-effective option to reduce tissue injury and preserve ovarian reserve.\u003c/p\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003eLimitations and Future Directions\u003c/h2\u003e \u003cp\u003eWhile this study provides valuable information on the effects of NAC in a rat model of ovarian torsion-detorsion, there are certain limitations. First, the study was conducted in an animal model, and while this provides valuable insights, further clinical studies are needed to translate these findings to human patients. Additionally, the mechanisms by which NAC influences asprosin expression and metabolic balance in ovarian tissues remain unclear and warrant further investigation. Finally, the duration of the torsion period and the use of NAC as a single dose may not fully replicate the clinical scenario, where delays in diagnosis and treatment are common. Therefore, future studies should explore different treatment regimens, timing of administration, and longer follow-up periods to better understand the clinical potential of NAC in ovarian torsion.\u003c/p\u003e \u003c/div\u003e"},{"header":"Conclusion","content":"\u003cp\u003eIn conclusion, this study demonstrates that NAC effectively reduces oxidative stress and mitigates the adverse metabolic effects associated with ovarian torsion-detorsion injury. By partially restoring asprosin levels and reducing TOS, NAC holds promise as a therapeutic agent for protecting ovarian tissue in cases of ischemia-reperfusion injury. While AMH levels remained unchanged, the significant biochemical and histological improvements observed in the NAC-treated group suggest a potential role for NAC in preserving ovarian reserve following torsion-detorsion events. Future research should focus on optimizing NAC treatment protocols and investigating its long-term effects in clinical settings.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cp\u003eThe following abbreviations are used in this manuscript:\u003c/p\u003e\n\u003cp\u003eT/D, torsion-detorsion;\u003c/p\u003e\n\u003cp\u003eTOS, total oxidant status;\u003c/p\u003e\n\u003cp\u003eROS, reactive oxygen species;\u003c/p\u003e\n\u003cp\u003eNAC, N-acetylcysteine;\u003c/p\u003e\n\u003cp\u003eAMH, anti-Müllerian hormone;\u003c/p\u003e\n\u003cp\u003eI/R, ischemia-reperfusion.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAuthor Contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eConceptualization, G.B., T.K. and A.A.; methodology, G.B, T.K and A.A.; software, C.S. and A.B.; validation, G.B; formal analysis, A.B. and C.S.; investigation, G.B., T.K. and A.A.; resources, G.B.; data curation, G.B. and A.A.; writing—original draft preparation, G.B.; writing—review and editing, G.B., A.B. and C.S.; visualization, A.B.; supervision, G.B and T.K.; project administration, G.B.; funding acquisition, G.B. All authors have read and agreed to the published version of the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research has not received any specific grants from funding organizations in the public, commercial or non-profit sectors.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eInformed Consent Statement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eInformed consent was obtained from all subjects involved in the study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability Statement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData are contained within the article.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflicts of Interest\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no conflicts of interest.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003eBedekar K, McInnes A, Burgess W. 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From imbalance to impairment: the central role of reactive oxygen species in oxidative stress-induced disorders and therapeutic exploration. Front Pharmacol. 2023; 14: 1269581. https://doi.org/10.3389/fphar.2023.1269581.\u003c/li\u003e\n \u003cli\u003eTang\u0026uuml;l, S. U., \u0026Ccedil;akmak, A. M., \u0026Ccedil;ağlayan, O., \u0026amp; Bozdoğan, \u0026Ouml;. (2020). Prevention of the harmful effects of free oxygen radicals by using N-acetylcysteine in testicular torsion. Journal of Pediatric Urology, 16(1), 42-e1. https://doi.org/10.1016/j.jpurol.2019.10.028\u003c/li\u003e\n \u003cli\u003eSandhu JK, et al. Oxidative Stress in Polycystic Ovarian Syndrome and the Effect of Antioxidant N-Acetylcysteine on Ovulation and Pregnancy Rate. Cureus. 2021 Sep 11;13(9): e17887. https://doi.org/10.7759/cureus.17887.\u003c/li\u003e\n \u003cli\u003eErsoy, G. S., Eken, M., Tal, R., Oztekin, D., Devranoglu, B., Kaygusuz, E. I., \u0026amp; Cevik, O. (2016). 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Cell Metab. 2022 Apr 5;34(4):549-563.e8. https://doi.org/10.1016/j.cmet.2022.02.012.\u003c/li\u003e\n \u003cli\u003eCorica D, et al. Asprosin serum levels and glucose homeostasis in children with obesity. Cytokine. 2021 Jun;142:155477. https://doi.org/10.1016/j.cyto.2021.155477.\u003c/li\u003e\n \u003cli\u003eHoffmann JG, Xie W, Chopra AR. Energy Regulation Mechanism and Therapeutic Potential of Asprosin. Diabetes. 2020 Apr;69(4):559-566. https://doi.org/10.2337/dbi19-0009.\u003c/li\u003e\n \u003cli\u003eHossein N, Mehdi M, Karim D, Mahdi M, Ghasemi E. Spirulina supplementation and circuit resistance training (CRT) reduce serum asprosin and appetite and improve energy balance in men with obesity and overweight. Hormones (Athens). 2025 Mar;24(1):23-31. https://doi.org/10.1007/s42000-024-00595-2.\u003c/li\u003e\n \u003cli\u003eFang YQ, et al. N-acetylcysteine supplementation improves endocrine-metabolism profiles and ovulation induction efficacy in polycystic ovary syndrome. J Ovarian Res. 2024 Oct 16;17(1):205. https://doi.org/10.1186/s13048-024-01528-8.\u003c/li\u003e\n \u003cli\u003eOlesen H\u0026Oslash;, et al. N-acetylcysteine protects ovarian follicles from ischemia-reperfusion injury in xenotransplanted human ovarian tissue. Hum Reprod. 2021 Jan 25;36(2):429-443. https://doi.org/10.1093/humrep/deaa291.\u003c/li\u003e\n \u003cli\u003eKacar E, et al. Asprosin-induced alterations in female rat puberty and reproductive hormonal profiles. Arch Physiol Biochem. 2025 Feb;131(1):24-32. https://doi.org/10.1080/13813455.\u003c/li\u003e\n \u003cli\u003eMoolhuijsen LME, Visser JA. Anti-Mullerian Hormone and Ovarian Reserve: Update on Assessing Ovarian Function. J Clin Endocrinol Metab. 2020 Nov 1;105(11):3361-73. https://doi.org/10.1210/clinem/dgaa513.\u003c/li\u003e\n \u003cli\u003eCedars MI. Evaluation of Female Fertility-AMH and Ovarian Reserve Testing. J Clin Endocrinol Metab. 2022 May 17;107(6):1510-1519. https://doi.org/10.1210/clinem/dgac039.\u003c/li\u003e\n \u003cli\u003eHarris BS, et al. Markers of ovarian reserve as predictors of future fertility. Fertil Steril. 2023 Jan;119(1):99-106. https://doi.org/10.1016/j.fertnstert.2022.10.014.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":true,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Ovarian torsion-detorsion, asprosin, N-acetylcysteine, ischemia-reperfusion, immunohistochemistry, rat model","lastPublishedDoi":"10.21203/rs.3.rs-6623339/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6623339/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eObjective:\u003c/strong\u003e Ovarian torsion is a life-threatening gynecological emergency that often necessitates surgical intervention. This study aimed to evaluate the effects of N-acetylcysteine (NAC), a potent antioxidant, on ovarian reserve and asprosin levels in a rat model of torsion-detorsion.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMaterials and Methods:\u003c/strong\u003e Thirty-five Wistar albino rats were randomly assigned to five groups: Group I (Control), Group II (Sham), Group III (NAC), Group IV (Torsion/Detorsion - T/D), and Group V (Torsion/Detorsion + NAC). Ovarian and blood samples were collected to assess asprosin immunoreactivity in tissue, serum asprosin levels, anti-Müllerian hormone (AMH) concentrations, and total oxidant status (TOS). Histopathological changes in ovarian tissues were evaluated using immunohistochemical techniques.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults:\u003c/strong\u003e Serum TOS levels were significantly elevated in the T/D group (p = 0.016) compared to the control group, and significantly decreased in the T/D + NAC group (p = 0.032) compared to the T/D group. Serum asprosin levels were significantly lower in the NAC (p = 0.002), T/D (p = 0.003), and T/D + NAC (p = 0.010) groups compared to the control (overall p = 0.016). However, asprosin levels were significantly higher in the T/D + NAC group than in the T/D group (p = 0.008). In ovarian tissue, asprosin immunoreactivity was significantly reduced in the T/D group compared to the control (p = 0.002), but increased in the T/D + NAC group compared to the T/D group (p = 0.002). No statistically significant differences were observed in AMH levels among the groups (p \u0026gt; 0.05).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion:\u003c/strong\u003e N-acetylcysteine (NAC) effectively reduced oxidative stress and prevented degenerative changes associated with ovarian torsion-detorsion injury. Asprosin levels appeared to reflect ischemia-reperfusion injury, decreasing with torsion and partially recovering with NAC treatment.\u003c/p\u003e","manuscriptTitle":"The Effects Of N-Acetylcysteine on Asprosine Immunoreactivity in A Rat Ovarian Torsion-Detorsion Model","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-05-14 09:28:14","doi":"10.21203/rs.3.rs-6623339/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"4d1fc523-af3d-4488-8048-a600c4fd2690","owner":[],"postedDate":"May 14th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-11-14T07:08:58+00:00","versionOfRecord":[],"versionCreatedAt":"2025-05-14 09:28:14","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6623339","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6623339","identity":"rs-6623339","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}