Effect of Dietary Patterns on Testosterone-Induced Prostatic Hyperplasia in Rats

Effect of Dietary Patterns on Testosterone-Induced Prostatic Hyperplasia in Rats | 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 Effect of Dietary Patterns on Testosterone-Induced Prostatic Hyperplasia in Rats Bernard Serebour, Samson Nwennajian Adongo, George Kwaw Ainooson, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6946787/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 Background: Benign prostatic hyperplasia (BPH) is one of the most serious urinary system disorders in elderly men worldwide. It is characterized by prostate enlargement and bladder outlet obstruction and several symptoms clinically referred to as lower urinary tract symptoms. Reported risk factors include advancing age, circulatory androgens, and genetics. Nevertheless, the role of Dietary patterns in the development of this condition remains largely unresolved. This study, therefore, investigated the influence of consumption of high fat from beef and pork source, high table sugar and high salt on Effect of Dietary Patterns on testosterone-induced prostatic hyperplasia in a rat model. Method : Thirty (30) male Sprague Dawley (SD) rats weighing 200 - 250g were randomly assigned to the pork fat, beef fat, high sugar (HSu), and high salt (HS) groups. After establishing the minimum threshold amount of testosterone above which significant prostate enlargement occurred, 50µg, all experimental groups were administered with the minimum testosterone in addition to the various dietary treatments for four weeks. Positive and negative control groups were administered with 100µg and 50µg testosterone respectively, in addition to a standard rat diet for four weeks. After the 28 days, the final weights of animals were recorded, animals sacrificed, blood samples taken for serum PSA determination, prostates and seminal vesicles isolated, weighed, prostate weight index (PI) determined, and prostate tissues processed for histopathological analysis. Mean ± SD values were calculated for each group and compared for significant difference using ANOVA followed by post hoc test (Dunn's Multiple Comparison Test) at p < 0.05. Results: Compared to the negative control, the mean serum PSA levels and PI of the high-fat and HSu groups were significantly higher, but they did not differ from those of the positive control. Although the mean PSA of the HS group was significantly higher compared to the negative control, the mean PI did not differ from the negative control. Conclusion: Collectively, our findings indicate that consumption of high-fat from red meat and high sugar diets enhance the effects of low testosterone to induce prostate enlargement in a rat model. BPH High fat diet (HFD) table sugar salt Prostatic Index (PI) Prostate Specific Antigen (PSA) hyperplasia prostate enlargement Figures Figure 1 Figure 2 Figure 3 Figure 4 Background Benign prostatic hyperplasia (BPH) is one of the most serious disorders of the urinary system in elderly men globally [ 1 ]. It accounts for substantial morbidity among elderly men and is generally not regarded as a preventable condition. It is characterized by the overgrowth of epithelial elements in the transitional zones of the peri- urethral area of the gland, leading to constriction of the prostatic urethra and bladder outlet obstruction (BOO), which together are commonly referred to as lower urinary tract symptoms (LUTS) [ 2 ]. If untreated, the condition can lead to bladder disease and impair kidney function [ 1 , 3 ]. In addition to the high economic burden that BPH poses on individuals and their families, it also considerably reduces the quality of life for affected men [ 4 ]. Alongside aging and hereditary factors, androgenic signaling serves as the primary stimulus for prostatic proliferation and BPH development. Furthermore, several emerging lines of evidence suggest that modifiable factors, such as diet, significantly contribute to the development of BPH and its treatment outcomes [ 4 – 6 ]. This is supported by epidemiological reports on the global incidence of BPH in various populations, which indicate a disparity in incidence between Asia (China, Japan, and India) and North America and Europe [ 7 – 10 ]. Among the major lifestyle differences between Asia and the West are their dietary habits and patterns [ 7 – 10 ]. The prevalence of BPH in sub-Saharan Africa (SSA) is considerably high, although it is grossly under-reported [ 11 – 13 ]. However, whether the increasing shift to Western dietary patterns and lifestyle in SSA contributes to the rising incidence of non-communicable diseases, including BPH, in SSA remains to be determined [ 14 , 15 ]. Preclinical and clinical studies have shown that the Western dietary pattern accelerates systemic alterations linked to enhanced genetic changes and increased cell proliferation [ 15 – 17 ]. Considering the global epidemic of diet-associated metabolic diseases, combined with the high frequency of newly diagnosed BPH and prostate cancers in developed and some developing countries, a better understanding of the role that components of diet, such as sugar, animal fats, and salts, play in the development of BPH is of significant importance. This preclinical study, therefore, aimed to elucidate the role of selected dietary components (or dietary patterns) in testosterone-dependent enlargement of the prostate, using a murine BPH model. Methodology Animal Husbandry. A total of Sixty (60) specific pathogen-free Sprague Dawley adult male rats weighing 200–250 g were obtained from the Faculty of Pharmacology and Pharmaceutical Sciences (FPPS-KNUST) for the study. The animals were habituated for two weeks in the Animal House of the same Faculty in an air-conditioned atmosphere, at a temperature of 22 ± 2°C with alternating 12-hour light and dark cycles throughout the experiment and provided with standard laboratory rat chow with water to drink ad libitum before the experiment began. Ethics Statement All experimental protocols and procedures were conducted in accordance with international ethical guidelines for the care and use of Laboratory Animals. Ethical approval was also obtained from the Animal Research Ethics Committee of the Kwame Nkrumah University of Science and Technology (KNUST/AREC/C.1–0076). Experimental design Beef and Pork Fat Extraction Beef and pork fats were purchased at a regional commercial butchery, washed with clean portable water, and boiled separately with a small amount of water under moderate heat until the water-free fats were obtained. The fats were stored in clean, labelled containers at -20 o C for later use. High Sugar and High Salts A 30% w/v sugar solution was prepared by dissolving 150g of table sugar (sucrose) in 500 mL of distilled water [ 18 ]. Likewise, a 4% w/v salt solution was prepared by dissolving 20 g salt (sodium chloride) in 500 mL distilled water to obtain a homogenous solution [ 19 ]. Establishment of the BPH Rat Model using testosterone Ten (10) Sprague Dawley (SD) adult male rats weighing 200–250 grams were randomly grouped into two groups of five and housed in clean aluminium cages. The first group was labelled Negative Control (NC) (administered with solvent only). The second group was labelled Testosterone ( T ) only (Testosterone manufacturer - FUJIFILM Wako Pure Chemical, Cat# 201-20551, Lot. CAL1859). The T -only group was subcutaneously administered 400 µg of T . All injections were administered at the same time of day, repeated every 72 hours for 28 days, as previously described by Mbaka et al. [ 20 ]. A successful induction of prostate enlargement was achieved with the administration of 400µg T for 28 days. However, the minimum dose of T above which resulted in a significant enlargement of the prostates needed to be ascertained, and to do that, a serial dilution of the 400µg T initially used was prepared: 200µg, 100µg, 50µg and 25µg, and administered to another set of SD rats of similar weight (200–250g) for 28 days. The animals were finally weighed, after which they were humanely sacrificed by euthanizing with isoflurane anesthesia at the end of the 28 days. The prostates and seminal vesicles were isolated and weighed. Whole blood of each rat was collected into a 15 ml Falcon tube by cardiac puncture and centrifuged at 3000 rpm for 15 min, and the serum obtained was separated and stored at -20°C for further analysis. Subsequently, the serum was used to estimate the total prostate-specific antigen (PSA) concentration. Animal grouping and dietary treatment. Another set of thirty (30) SD adult male rats of similar weight (200–250 g) were obtained and randomly assigned to six groups of five, marked and housed in different cages. The groups were labelled as follows: negative control (NC), positive control (PC), High salt (HS), High sugar (HSu), Beef fat, and Pork fat groups. The NC group rats were subcutaneously administered with 0.5 ml of the solvent (anise oil only) every 72 hours for the entire four-week period. The PC group was similarly administered with 100µg T every 72 hours for four weeks as described by Mbaka et al . (2017). The HS group was given 4ml of 4% salt solution every 12 hours for the entire duration of the study. In addition, 50µg T was administered subcutaneously every 72 hours for 28 days. The HSu group was also orally administered with 4ml of 30% w/v sucrose solution every 12-hour day for the entire duration of the study. In addition, 50µg T was subcutaneously administered every 72 hours for the 28-day period of the study. The extracted fats were pre-warmed to 30°C in a water bath before orally administered to the rats in the Beef and Pork fat groups. Each rat was given 2 ml of the respective fat orally each day for the entire duration of the experiment. Rats in these groups were also administered 50µg T every 72 hours for the four-week duration of the study. All rat groups had unhindered access to standard rat chow and water to drink ad libitum. After final treatment and overnight fasting, the rats were humanely sacrificed by euthanizing with isoflurane anesthesia. Whole blood of each rat was collected into 15 ml Falcon tubes by cardiac puncture and centrifuged at 3000 rpm for 15 min, after which the serum was separated and stored at -20°C. Prostate tissues were also isolated and immediately weighed. Assessment of Prostate Weight (PW) and Prostatic Weight Index (PI) The final body weight (BW) and prostate weight (PW) were determined for all rat groups as well as their respective means. Prostate Index (PI) was determined from the equation \(\:\frac{PW}{BW}\times\:100\%\) and the mean PI were calculated for every group. The mean percentage increase in PI was also calculated from the PI of the controls and the diet treatment groups ( \(\:\frac{diet\:treated\:PI-control\:PI}{Control\:PI}\times\:100\%)\) Serum Prostate-Specific Antigen (PSA) Estimation. Total serum PSA was estimated using PSA Enzyme Immunoassay test kit according to the manufacturer’s protocol. A microplate reader (iMark™, Bio-Rad Laboratories, USA) was used to read the optical density at 450nm within 15 minutes of colour development. Histopathological examination . Sections from prostate tissues were fixed in 10% neutral buffered formalin and processed for paraffin sections of 4 µm thickness. The sections were collected on glass slides, deparaffinized then stained with haematoxylin and eosin (H&E) for routine histopathological examination. Histopathological slides were analysed by an expert pathologist with knowledge in murine pathology, who was blind to the experimental conditions. A light microscope (Olympus CX23 Binocular Microscope) was used for image processing. The prostate tissue histology was evaluated by comparing the treated groups to those of the controls to reveal histological changes. Statistical Analysis All data were expressed as mean ± standard error of mean (SEM), and comparison of mean parameters measured between experimental groups and control groups were determined using One-way ANOVA (Kruskal-Wallis test and Dunn's Multiple Comparison Test) using GraphPad Prism version 8.0.2. P ˂ 0.05 was considered statistically significant. Results Androgen signalling remains a major driver of prostate cell proliferation and BPH development; although serum testosterone levels decrease with advancing age (by about 2–3% per year), the risk of developing BPH also increases with advancing age. This suggests that other factors enhance low serum testosterone to induce enlargement of the prostate, leading to BPH. In this study, we assessed the influence of four dietary components on the hormone-dependent enlargement of the prostate. Minimum Testosterone Above Which BPH Induction Occurs . To assess the potentiating effect of the dietary substances, the minimum amount of testosterone ( T) above which significant enlargement of the prostate (BPH induction) occurred was determined. This was done by serially diluting the 400µg T into 200µg, 100µg, 50µg and 25µg and administering to obtain the minimum dose of 50µg T above which BPH induction occurred (Fig. 1 a – 1 c). Subsequently, 100µg Testosterone was used as the positive control, while 50µg Testosterone was adopted as the minimum hormone to be administered concurrently with the dietary treatments to assess their potentiating effects. High Fat Diet (Pork and Beef sources) Increased Prostate Weight Index and Serum PSA. Rats fed with a high-fat diet from pork and beef, respectively, with the minimum determined testosterone (50µg) dose showed significantly greater average PI of 2.56 ± 0.08g and 2.35 ± 0.06g compared with 1.30 ± 0.0 g for the 50µg treated only. Similarly, the serum PSA levels in the HFD groups were significantly higher compared with the 50µg treated only (Table 1 ). Compared with the 50µg treated only, there were 96.3% and 80.2% increases in PI for pork and beef fat groups, respectively. The percentage increase in serum PSA for the pork and beef fat-fed rats respectively were 72.3% and 77.2% compared to the 50µg treated only. However, there was no significant difference in the PI and PSA levels induced by the pork fat-fed compared with the beef fat-fed groups. Table 1: Percentage increase in PSA levels after Testosterone and dietary treatment Table 1 High-fat diet enhanced low testosterone concentration to induce elevation of serum PSA. Data are expressed as mean ± SEM (n = 5). A one-way ANOVA was carried out to compare the minimum testosterone (50µg only) with the positive control (100µg T), 50µg T + Beef fat and 50µg T + Pork fat groups. * p-value < 0.05. High Pork and Beef Fat Diet Promote Prostate Enlargement under Low Testosterone in Rats Next, we examined the effect of intake of high pork and high beef fat diets with the minimum T on the prostate histology by performing a histopathological examination. Changes in the prostate tissues were examined after H&E staining. The prostatic epithelial cells in the solvent control group were cuboidal and regular in size (Fig. 4 ). Nevertheless, the epithelial cell layer in the positive control and the fat-fed rats were larger, and the epithelium became hyperplastic. Numerous papillae projecting into the lumen formed and further decreased the glandular luminal area (Fig. 4 ). In the pork and beef fat diet groups, visible changes were observed showing much thickened epithelia and acini, hypertrophy, wider diameter of glandular lumen and hyperplasia with less marked inter-glandular stroma (Fig. 4 ). There were no alterations in the histological appearance of the epithelial tissue linings in the solvent control. Thin layers of epithelia were observed, indicative of no induced hyperplasia. High Salt and Sugar Diets slightly increased Prostate Index, serum PSA, and altered the Histology of Prostatic Tissues in BPH rats In comparison with the control group (minimum T only), rats on a high-salt diet plus minimum T showed no significant increase in PI and serum PSA levels, however, rats on the high-sugar diet plus 50µg T showed significantly higher PI (1.74 ± 0.15) compared to the minimum T only control (1.30 ± 0.04). Similarly, the serum PSA levels (0.64 ± 0.01) was significantly higher compared with the 50µg T only treated (Fig. 3 b). Significant increase in the levels of serum PSA was observed in high sugar treated group (0.64 ± 0.01 ng/ml) compared to the control (0.46 ± 0.01 ng/ml) (Table 3). In the HSu diet group, the increased PI and elevated serum PSA levels were accompanied by hyperplasia of glandular epithelium and inter-glandular stroma, which was absent in the control group, that showed thin glandular epithelia and fibro-muscular layers (Fig. 4 ). However, the rats fed with HS displayed features similar to that of the 50µg T only control (Fig. 4 ). Table 2 Serum PSA levels after Testosterone and Dietary treatments. Group (n = 5) PSA (ng/ml) % Increase Solvent control 0.43 ± 0.01 50µg T only (minimum testosterone) 0.46 ± 0.01 6.90 100µg T only 0.75 ± 0.05** 74.42 50µg T + High Salt 0.47 ± 0.03 9.30 50µg T + High Sugar 0.64 ± 0.01* 48.83 Table 2: High-sugar diet but not high-salt diet enhanced low testosterone concentration to induce the elevation of serum PSA. Data are expressed as mean ± SEM (n = 5). A one-way ANOVA was carried out to compare the minimum testosterone (50µg only) with the positive control (100µg T), 50µg T + High Sugar and 50µg T + High Salt groups. * p-value < 0.05. Histopathological Examination . Microscopic examination of H&E-stained sections of the ventral prostatic tissues taken from solvent control and the 50µg T only group showed normal structure in the form of numerous acini of different sizes and shapes containing homogenous acidophilic material. The lining epithelium of the acini was composed of two distinct cell types, namely, basal and acinar or principal cells. The acinar cells were columnar, extending from the basal lamina to the lumen of the duct (Fig. 4 ). Conversely, prostates obtained from experimental groups C, D, E, F, and G rats displayed an obvious disruption of the histoarchitecture of the prostatic tissue shown as marked hyperplasia, irregular acinar folding with intraluminar projections, and congestion (Fig. 4 ). However, similar to the prostate tissues from the positive control (C), glands with prominent widespread epithelial hyperplasia (j) and fibromuscular stromal hypertrophy (s) were identified in the experimental groups (Fig. 3 . D, E and G) Discussion BPH is considered the most common urological disorder affecting men above 50 years of age with a prevalence of up to 85% in older age groups [ 1 ]. It is characterized by non-cancerous enlargement and uncontrolled proliferation of smooth muscles and epithelial cells of the prostatic tissue. Clinically, BPH is manifested as LUTS with a significant negative impact on the quality of life [ 21 ]. While earlier reports indicate that the development and progression of BPH are dependent on several factors [ 22 ], there is not enough clarity on the role of dietary patterns in the development of this condition. In this study, we report that chronic intake of high-fat from red meat sources in the presence of low testosterone induces enlargement of the prostate in a rat model. Our data suggests that high-fat diet enhances low concentration of T , which in itself, is not sufficient to induce prostate enlargement (hyperplasia) in the initiation of a BPH disease. Increase in PI and histological alterations in the prostate tissues have been important parameters in assessing prostate health and BPH development [ 23 , 24 ]. HFD consumption has been strongly associated with higher BPH risk and potentially, cancer of the prostate (PCa) in several studies [ 5 , 8 , 17 ]. How red meat fats enhanced low T action to induce enlargement in this study is not fully understood, however, it has been reported by earlier studies that, HFD contain high amounts of saturated fatty acids (SFA) [ 25 , 26 ] which plays a role in inflammation and enhances the production of reactive oxygen species (ROS) which worsens the oxidative stress symptoms in the prostate and as a consequence, hyperplasia is enhanced as a developmental adaptation under these conditions [ 27 ]. Another report consistent with ours was published by Shankar and colleagues, which indicated that an NF-κB reporter mice fed a high-fat diet (HFD) with a composition characteristic of a Western-style diet resulted in increased intra-prostatic inflammation and cell proliferation, leading to prostate gland enlargement [ 8 , 28 ]. A recent report has indicated that HFD profoundly alters the expression of pro-proliferative genes and even an oncogene, such as MYC, resulting in the enhancement of MYC-driven transcriptional programs in prostate cells, leading to increased glandular cell proliferation and potential transformation of the prostate, in the case of cancer [ 17 ]. These aforementioned reports support our findings that, even though excessive intake of animal fat is considered generally harmful to the body, it could also enhance prostate glandular cell proliferation under low T levels in the development of enlarged prostate in men. In this study, we report that chronic consumption of high sugar enhanced low testosterone levels to induce enlargement of the prostate in rats. The reason for the observed synergy is not clearly decipherable. However, earlier studies have reported a strong association between fasting blood sugar and the size of the prostate [ 29 ]. Mechanistically, androgen treatment-induced prostatic hyperplasia in vivo is dependent on increased expression of growth factors, mainly IGF-1, and its receptor IGF-1R, and increased expression of phosphorylated Akt as well as Bcl-2 [ 30 ]. It also significantly decreases testosterone–induced Bax / Bcl-2 ratio and P 21 activity, thereby inhibiting apoptosis [ 31 ]. This may be one mechanism by which glucose enhances the action of low testosterone in promoting prostatic hyperplasia. Secondly, others have reported that high glucose treatment could induce proliferation and suppress apoptosis in prostatic epithelial cells through a different mechanism [ 32 ]. Ye et al. (2011, 2018) found that under a high glucose environment, the level of reactive oxygen species (ROS) was significantly up-regulated, which induced cell proliferation and suppressed the apoptosis of prostate epithelial cells [ 33 , 34 ]. Taken together, our findings suggest that high glucose treatment might facilitate BPH development, even in the presence of low testosterone. Average salt consumption in humans ranges from 9 to 12g per day (double the recommended dose), of which most of it is found in food condiments [ 35 ]. It is known that high-salt (> 2g/day), which constitutes 40% of sodium intake, is a huge underlying risk factor for elevated arterial blood pressure, heart diseases, hypertension, and stroke [ 8 , 28 ]. High serum sodium has been associated with the development of BPH and worse lower urinary tract symptoms [ 8 , 28 ]. In our studies, a high salt diet over the period of 28 days did not enhance the minimum testosterone dose to increase the PI and therefore did not significantly differ when compared with that of NC. However, there were histological changes that appeared somewhat similar to the alterations observed in the positive control, as well as a slight increase in mean serum PSA, which could indicate an insidious inflammation flourishing in the prostate gland in response to the high salt intake. The mechanism by which high salt enhanced low T action to cause the increased serum PSA of the HS rats' prostate is unclear. Salt is known to aggravate the symptoms of BPH and LUTS through the bladder and prostate innervation. In terms of mechanism, it is generally believed that it may be related to abnormal activation of the sympathetic nervous system [ 8 , 28 ]. However, findings consistent with what we observed in this study was reported by Bello and colleagues (2023), who showed that low, standard, and high salt diets aggravated the pathology of testosterone-induced BPH in Wistar rats by promoting inflammation, oxidative stress, while suppressing apoptosis and angiogenesis [ 36 ]. Although an insignificant enlargement of prostate glands was observed in the high salt group in this study, a significant increase in serum PSA levels was observed in the group compared with the controls. Therefore, the effect of high intake of salt observed in this study could be explained by pro-inflammatory activity of high dietary salt and its potential to enhance low T to aggravate BPH pathophysiology by increasing epithelial thickness and polyps formation, which may have consequently raised the serum PSA levels. To the best of our knowledge, this study provides insight into the effect of dietary components and low testosterone on the risk of BPH in a rat model. The study showed the synergistic effect of high intake of red meat fat, table sugar, and table salt and their risk of BPH. However, there are some potential limitations of this study. Firstly, our study did not evaluate inflammatory cytokines in the prostate, as well as, gene expression profile of genes known to promote hyperplasia in the prostate, which would have provided more insight into the mechanism by which these dietary components enhance testosterone-mediated gene expression in the prostate. Conclusion This study demonstrated that rat diets high in fat from red meat, high in sugar, and high in dietary salt promoted hyperplasia and enlargement of prostatic tissues under low testosterone levels. Therefore, it is concluded that dietary fats, sugars, and, to a lesser extent, salt intake enhanced the development of testosterone-induced BPH in rats. These observations suggest that dietary patterns are a contributing risk factor for BPH development. We recommend further molecular mechanistic studies to help clarify the causal role of various dietary patterns in the initiation or induction of BPH. Declarations Ethics, Consent to Participate, and Consent to Publish declarations. Ethics Statement All experimental protocols and procedures were carried out in accordance with international ethical guidelines for the care and use of Laboratory Animals. Ethical approval was also obtained from the Animal Research Ethics Committee of the Kwame Nkrumah University of Science and Technology (KNUST/AREC/C.1- 0076). Consent for publications Not applicable. Our manuscript contains data generated from original research involving animal model study; therefore, it does not contain any person’s data. Availability of data and materials The datasets used and/or analyzed during the current study are available from the corresponding author upon reasonable request. Competing interests The authors declare that they have no conflicts of interest. Funding This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors. Authors’ contributions BS: investigation, methodology, and writing original draft; SA: investigation, methodology; KAAA , GKA: supervision, review & editing; EAN: conceptualization, project administration, resources, supervision, investigation, methodology, formal analysis, writing, review & editing. All the authors read and approved the final manuscript. Acknowledgments The authors are very grateful to the Histopathologist, Mr. Clement Atobiga, and the entire staff at the Department of Pharmacology Animal House for their support in this study. 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The effect of Metapanax delavayi leaf extract on testosterone-induced benign prostatic hyperplasia in rats. J Funct Foods 2020;66. https://doi.org/10.1016/j.jff.2020.103797 Isabel, et al. Western Diet: Implications for Brain Function and Behavior. Front Psychol. 2020;11:1–11. https://doi.org/10.3389/fpsyg.2020.564413 . Maserejian NN, Giovannucci EL, McKinlay JB. Dietary Macronutrients, Cholesterol, and Sodium and Lower Urinary Tract Symptoms in Men. Eur Urol. 2009;55:1179–89. https://doi.org/10.1016/j.eururo.2008.07.058 . Kitamoto K, Miura Y, Karnan S, Ota A, Konishi H, Hosokawa Y, et al. Inhibition of NADPH oxidase 2 induces apoptosis in osteosarcoma: The role of reactive oxygen species in cell proliferation. Oncol Lett. 2018;15:7955–62. https://doi.org/10.3892/ol.2018.8291 . Shankar E, Bhaskaran N, MacLennan GT, Liu G, Daneshgari F, Gupta S. Inflammatory Signaling Involved in High-Fat Diet Induced Prostate Diseases. J Urol Res. 2015;2:1–18. Kim WT, Yun SJ, Choi YD, Kim GY, Moon SK, Choi YH, et al. Prostate size correlates with fasting blood glucose in non- diabetic benign prostatic hyperplasia patients with normal testosterone levels. J Korean Med Sci. 2011;26:1214–8. https://doi.org/10.3346/jkms.2011.26.9.1214 . Mosli HH, Esmat A, Atawia RT, Shoieb SM, Mosli HA, Abdel-Naim AB. Metformin Attenuates Testosterone-Induced Prostatic Hyperplasia in Rats: A Pharmacological Perspective. Sci Rep. 2015;5:1–10. https://doi.org/10.1038/srep15639 . Clemmons DR. Role of Insulin-Like Growth Factor Iin Maintaining Normal Glucose Homeostasis. Horm Res Paediatr. 2004;62:77–82. https://doi.org/10.1159/000080763 . Wei P, Lin D, Luo C, Zhang M, Deng B, Cui K, et al. High glucose promotes benign prostatic hyperplasia by downregulating PDK4 expression. Sci Rep. 2023;13:17910. https://doi.org/10.1038/s41598-023-44954-2 . Ye C, Cai Y, Cai Q, Yuan S, Huang F, Yang X, et al. High glucose induces the proliferation of prostatic cells via downregulating MRE11. Int J Mol Med. 2018. https://doi.org/10.3892/ijmm.2018.3549 . Ye C, Li X, Wang Y, Zhang Y, Cai M, Zhu B, et al. Diabetes causes multiple genetic alterations and downregulates expression of DNA repair genes in the prostate. Lab Investig. 2011;91:1363–74. https://doi.org/10.1038/labinvest.2011.87 . He FJ, Brown M, Tan M, MacGregor GA. Reducing population salt intake—An update on latest evidence and global action. J Clin Hypertens. 2019;21:1596–601. https://doi.org/10.1111/jch.13664 . Bello II, Omigbodun A, Morhason-Bello I. Common salt aggravated pathology of testosterone-induced benign prostatic hyperplasia in adult male Wistar rat. BMC Urol. 2023;23:207. https://doi.org/10.1186/s12894-023-01371-x . Additional Declarations No competing interests reported. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6946787","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":495101716,"identity":"99cfb767-d205-4a15-a884-0cd4eaa39338","order_by":0,"name":"Bernard Serebour","email":"","orcid":"","institution":"Kwame Nkrumah University of Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Bernard","middleName":"","lastName":"Serebour","suffix":""},{"id":495101717,"identity":"d093b930-0f9b-4016-95f7-82cd5e5c62b2","order_by":1,"name":"Samson Nwennajian Adongo","email":"","orcid":"","institution":"Kwame Nkrumah University of Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Samson","middleName":"Nwennajian","lastName":"Adongo","suffix":""},{"id":495101718,"identity":"e76472e8-bd48-4468-8d3a-5ca985294495","order_by":2,"name":"George Kwaw Ainooson","email":"","orcid":"","institution":"Kwame Nkrumah University of Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"George","middleName":"Kwaw","lastName":"Ainooson","suffix":""},{"id":495101719,"identity":"9b2aa9e3-b33d-4287-9060-940927287be2","order_by":3,"name":"Kwaku Addai Arhin Appiah","email":"","orcid":"","institution":"Kwame Nkrumah University of Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Kwaku","middleName":"Addai Arhin","lastName":"Appiah","suffix":""},{"id":495101720,"identity":"e8f11f7e-fef8-4e8b-9049-7c4e2d25779c","order_by":4,"name":"Emmanuel Amankwah Ntim","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA/klEQVRIiWNgGAWjYLCCBCA2YGA+DOMbAPEBfBoYGyBa2JJBPAnitEBM5jEmTgt/e+/xBw9zGOTN+c98Ni5sq6tjYG/eJsHw6w5OLRJnziU2JG5jMNw5I3dz8sy2wxIMPMfKJBj7nuF21o0cQ5CWBIMbvJsP87YdkGCQyDGTYOw5jFOH/P03UC3nzzwGaqmTYJB/g1+LwQ0eqJYDOczJvG3MQFt4zCQYfuDWYngmx3BG4jYJww030oyNec4dlmzjSSu2SGzArUXu+BmDjz+32cgbnD/8WJqnrI6fn/3wxhsf/uDWAgUSCCYbiEhsI6QDE/whXcsoGAWjYBQMWwAAOSpVIOxslQUAAAAASUVORK5CYII=","orcid":"","institution":"Kwame Nkrumah University of Science and Technology","correspondingAuthor":true,"prefix":"","firstName":"Emmanuel","middleName":"Amankwah","lastName":"Ntim","suffix":""}],"badges":[],"createdAt":"2025-06-21 20:53:08","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6946787/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6946787/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":88329503,"identity":"2e09e086-16cd-43c5-a110-3bc18d00170c","added_by":"auto","created_at":"2025-08-05 10:29:49","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":878716,"visible":true,"origin":"","legend":"\u003cp\u003e(a) A photograph of isolated prostates and seminal vesicles showing induction of enlargement using 400 μg testosterone compared to a solvent control. (b) Isolated prostates and seminal vesicles showing the minimum dose of testosterone above which there is an induction of enlargement of the prostate and seminal vesicles. Based on the serial dilution used, 50 µg was the minimum testosterone above which there was clear induction of hormone-dependent enlargement. Solvent: injected with vehicle only; \u003cem\u003e\u003cstrong\u003eT\u003c/strong\u003e\u003c/em\u003e - Testosterone administered groups: 25µg, 50µg, 100µg and 200µg of testosterone, respectively, every 72 hours for four weeks. (c) Prostate-weight index of testosterone-induced BPH groups compared to the solvent control. Data are expressed as mean ± SEM (n = 5). +25µg \u003cem\u003e\u003cstrong\u003eT\u003c/strong\u003e\u003c/em\u003e and +50 µg \u003cem\u003e\u003cstrong\u003eT\u003c/strong\u003e\u003c/em\u003e\u003cem\u003e \u003c/em\u003enot statistically significant from the solvent control. +100µg \u003cem\u003e\u003cstrong\u003eT \u003c/strong\u003e\u003c/em\u003eand 200µg \u003cem\u003e\u003cstrong\u003eT\u003c/strong\u003e\u003c/em\u003e statistically significant from solvent control group, at \u003cem\u003eP\u003c/em\u003e\u0026lt; 0.01 using one-way ANOVA (Kruskal-Wallis test and Dunn's Multiple Comparison Test).\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-6946787/v1/14fd03dd825fb58bb6e2cc34.png"},{"id":88328743,"identity":"ef7c32b3-4921-457c-ac15-fa556f38ad5e","added_by":"auto","created_at":"2025-08-05 10:21:49","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":444190,"visible":true,"origin":"","legend":"\u003cp\u003e(a) A photograph showing the prostates and seminal vesicles of Testosterone treated groups: 50µg, 100µg, 50µg \u003cem\u003e\u003cstrong\u003eT\u003c/strong\u003e\u003c/em\u003e + Pork fat, and 50µg + Beef fat. \u003cstrong\u003e(b)\u003c/strong\u003e Prostate-weight indices of Testosterone treated groups: 50µg, 100µg, 50µg \u003cem\u003e\u003cstrong\u003eT\u003c/strong\u003e\u003c/em\u003e + Pork fat, and 50µg + Beef fat. Data are expressed as mean ± SEM (n = 5). \u003cem\u003eP\u003c/em\u003e \u0026lt; 0.01 using one-way ANOVA.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-6946787/v1/bfa14420620bbe5fe29f8621.png"},{"id":88328412,"identity":"64d7e7e9-aebb-49e1-8974-8ca76475ad79","added_by":"auto","created_at":"2025-08-05 10:13:49","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":526270,"visible":true,"origin":"","legend":"\u003cp\u003e(a) A picture showing the prostate and seminal vesicles from Testosterone-induced BPH groups: 50µg testosterone, 100µg testosterone, 50µg \u003cstrong\u003eT \u003c/strong\u003e+ High salt, 50µg\u003cstrong\u003eT\u003c/strong\u003e + High sugar. (b) A figure showing the comparison of Prostate-weight index of the groups compared with the controls. Data are expressed as mean ± SEM (n = 5). \u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05 using one-way ANOVA. HSu – high sugar solution, HS – high salt solution, \u003cstrong\u003eT\u003c/strong\u003e - testosterone.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-6946787/v1/c80263a233594c075b837e44.png"},{"id":88328414,"identity":"5e51a03f-1e0c-4aea-a3a7-2568635f3637","added_by":"auto","created_at":"2025-08-05 10:13:49","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":1755993,"visible":true,"origin":"","legend":"\u003cp\u003eHematoxylin and Eosin (H\u0026amp;E) stained micrographs of prostate sections showing the influence of dietary components on testosterone-induced hyperplasia of the prostate. (A) Section from solvent control rats. (B)Section from 50µg testosterone only rats. (C) Section from the 100µg testosterone only treated group (positive control). (D) Section from 50µg testosterone plus beef fat; (E) Section from 50µg testosterone plus pork fat; (F), 50µg testosterone plus high salt diet and (G) 50µg testosterone plus high sugar diet (Images taken at 40x and 100x magnifications).\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-6946787/v1/e0b7009c545fb5467eb986ae.png"},{"id":90535563,"identity":"a47ab20c-1651-4d4c-9ffd-0a6aec40d1f4","added_by":"auto","created_at":"2025-09-03 20:01:59","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":4977297,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6946787/v1/a316eb3b-3161-4426-b4be-aeb35c169c42.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Effect of Dietary Patterns on Testosterone-Induced Prostatic Hyperplasia in Rats","fulltext":[{"header":"Background","content":"\u003cp\u003eBenign prostatic hyperplasia (BPH) is one of the most serious disorders of the urinary system in elderly men globally [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. It accounts for substantial morbidity among elderly men and is generally not regarded as a preventable condition. It is characterized by the overgrowth of epithelial elements in the transitional zones of the peri- urethral area of the gland, leading to constriction of the prostatic urethra and bladder outlet obstruction (BOO), which together are commonly referred to as lower urinary tract symptoms (LUTS) [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. If untreated, the condition can lead to bladder disease and impair kidney function [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. In addition to the high economic burden that BPH poses on individuals and their families, it also considerably reduces the quality of life for affected men [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. Alongside aging and hereditary factors, androgenic signaling serves as the primary stimulus for prostatic proliferation and BPH development. Furthermore, several emerging lines of evidence suggest that modifiable factors, such as diet, significantly contribute to the development of BPH and its treatment outcomes [\u003cspan additionalcitationids=\"CR5\" citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. This is supported by epidemiological reports on the global incidence of BPH in various populations, which indicate a disparity in incidence between Asia (China, Japan, and India) and North America and Europe [\u003cspan additionalcitationids=\"CR8 CR9\" citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. Among the major lifestyle differences between Asia and the West are their dietary habits and patterns [\u003cspan additionalcitationids=\"CR8 CR9\" citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. The prevalence of BPH in sub-Saharan Africa (SSA) is considerably high, although it is grossly under-reported [\u003cspan additionalcitationids=\"CR12\" citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. However, whether the increasing shift to Western dietary patterns and lifestyle in SSA contributes to the rising incidence of non-communicable diseases, including BPH, in SSA remains to be determined [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e, \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. Preclinical and clinical studies have shown that the Western dietary pattern accelerates systemic alterations linked to enhanced genetic changes and increased cell proliferation [\u003cspan additionalcitationids=\"CR16\" citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. Considering the global epidemic of diet-associated metabolic diseases, combined with the high frequency of newly diagnosed BPH and prostate cancers in developed and some developing countries, a better understanding of the role that components of diet, such as sugar, animal fats, and salts, play in the development of BPH is of significant importance. This preclinical study, therefore, aimed to elucidate the role of selected dietary components (or dietary patterns) in testosterone-dependent enlargement of the prostate, using a murine BPH model.\u003c/p\u003e"},{"header":"Methodology","content":"\u003cp\u003e\u003cb\u003eAnimal Husbandry.\u003c/b\u003e A total of Sixty (60) specific pathogen-free Sprague Dawley adult male rats weighing 200\u0026ndash;250 g were obtained from the Faculty of Pharmacology and Pharmaceutical Sciences (FPPS-KNUST) for the study. The animals were habituated for two weeks in the Animal House of the same Faculty in an air-conditioned atmosphere, at a temperature of 22\u0026thinsp;\u0026plusmn;\u0026thinsp;2\u0026deg;C with alternating 12-hour light and dark cycles throughout the experiment and provided with standard laboratory rat chow with water to drink \u003cem\u003ead libitum\u003c/em\u003e before the experiment began.\u003c/p\u003e\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003eEthics Statement\u003c/h2\u003e\u003cp\u003e All experimental protocols and procedures were conducted in accordance with international ethical guidelines for the care and use of Laboratory Animals. Ethical approval was also obtained from the Animal Research Ethics Committee of the Kwame Nkrumah University of Science and Technology (KNUST/AREC/C.1\u0026ndash;0076).\u003c/p\u003e\u003c/div\u003e\n\u003ch3\u003eExperimental design\u003c/h3\u003e\n\u003cp\u003e\u003cstrong\u003eBeef and Pork Fat Extraction\u003c/strong\u003e\u003cp\u003e Beef and pork fats were purchased at a regional commercial butchery, washed with clean portable water, and boiled separately with a small amount of water under moderate heat until the water-free fats were obtained. The fats were stored in clean, labelled containers at -20\u003csup\u003eo\u003c/sup\u003eC for later use.\u003c/p\u003e\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eHigh Sugar and High Salts\u003c/strong\u003e\u003cp\u003eA 30% w/v sugar solution was prepared by dissolving 150g of table sugar (sucrose) in 500 mL of distilled water [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. Likewise, a 4% w/v salt solution was prepared by dissolving 20 g salt (sodium chloride) in 500 mL distilled water to obtain a homogenous solution [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e].\u003c/p\u003e\u003c/p\u003e\n\u003ch3\u003eEstablishment of the BPH Rat Model using testosterone\u003c/h3\u003e\n\u003cp\u003eTen (10) Sprague Dawley (SD) adult male rats weighing 200\u0026ndash;250 grams were randomly grouped into two groups of five and housed in clean aluminium cages. The first group was labelled Negative Control (NC) (administered with solvent only). The second group was labelled Testosterone (\u003cb\u003eT\u003c/b\u003e) only (Testosterone manufacturer - FUJIFILM Wako Pure Chemical, Cat# 201-20551, Lot. CAL1859). The \u003cb\u003eT\u003c/b\u003e\u003cem\u003e-only\u003c/em\u003e group was subcutaneously administered 400 \u0026micro;g of \u003cb\u003eT\u003c/b\u003e. All injections were administered at the same time of day, repeated every 72 hours for 28 days, as previously described by Mbaka \u003cem\u003eet al.\u003c/em\u003e [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eA successful induction of prostate enlargement was achieved with the administration of 400\u0026micro;g \u003cb\u003eT\u003c/b\u003e for 28 days. However, the minimum dose of \u003cb\u003eT\u003c/b\u003e above which resulted in a significant enlargement of the prostates needed to be ascertained, and to do that, a serial dilution of the 400\u0026micro;g \u003cb\u003eT\u003c/b\u003e initially used was prepared: 200\u0026micro;g, 100\u0026micro;g, 50\u0026micro;g and 25\u0026micro;g, and administered to another set of SD rats of similar weight (200\u0026ndash;250g) for 28 days. The animals were finally weighed, after which they were humanely sacrificed by euthanizing with isoflurane anesthesia at the end of the 28 days. The prostates and seminal vesicles were isolated and weighed. Whole blood of each rat was collected into a 15 ml Falcon tube by cardiac puncture and centrifuged at 3000 rpm for 15 min, and the serum obtained was separated and stored at -20\u0026deg;C for further analysis. Subsequently, the serum was used to estimate the total prostate-specific antigen (PSA) concentration.\u003c/p\u003e\u003cp\u003e\u003cb\u003eAnimal grouping and dietary treatment.\u003c/b\u003e Another set of thirty (30) SD adult male rats of similar weight (200\u0026ndash;250 g) were obtained and randomly assigned to six groups of five, marked and housed in different cages. The groups were labelled as follows: negative control (NC), positive control (PC), High salt (HS), High sugar (HSu), Beef fat, and Pork fat groups.\u003c/p\u003e\u003cp\u003eThe NC group rats were subcutaneously administered with 0.5 ml of the solvent (anise oil only) every 72 hours for the entire four-week period. The PC group was similarly administered with 100\u0026micro;g \u003cb\u003eT\u003c/b\u003e every 72 hours for four weeks as described by Mbaka \u003cem\u003eet al\u003c/em\u003e. (2017). The HS group was given 4ml of 4% salt solution every 12 hours for the entire duration of the study. In addition, 50\u0026micro;g \u003cb\u003eT\u003c/b\u003e was administered subcutaneously every 72 hours for 28 days. The HSu group was also orally administered with 4ml of 30% w/v sucrose solution every 12-hour day for the entire duration of the study. In addition, 50\u0026micro;g \u003cb\u003eT\u003c/b\u003e was subcutaneously administered every 72 hours for the 28-day period of the study. The extracted fats were pre-warmed to 30\u0026deg;C in a water bath before orally administered to the rats in the Beef and Pork fat groups. Each rat was given 2 ml of the respective fat orally each day for the entire duration of the experiment. Rats in these groups were also administered 50\u0026micro;g \u003cb\u003eT\u003c/b\u003e every 72 hours for the four-week duration of the study. All rat groups had unhindered access to standard rat chow and water to drink \u003cem\u003ead libitum.\u003c/em\u003e After final treatment and overnight fasting, the rats were humanely sacrificed by euthanizing with isoflurane anesthesia. Whole blood of each rat was collected into 15 ml Falcon tubes by cardiac puncture and centrifuged at 3000 rpm for 15 min, after which the serum was separated and stored at -20\u0026deg;C. Prostate tissues were also isolated and immediately weighed.\u003c/p\u003e\n\u003ch3\u003eAssessment of Prostate Weight (PW) and Prostatic Weight Index (PI)\u003c/h3\u003e\n\u003cp\u003eThe final body weight (BW) and prostate weight (PW) were determined for all rat groups as well as their respective means. Prostate Index (PI) was determined from the equation \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\frac{PW}{BW}\\times\\:100\\%\\)\u003c/span\u003e\u003c/span\u003e and the mean PI were calculated for every group. The mean percentage increase in PI was also calculated from the PI of the controls and the diet treatment groups (\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\frac{diet\\:treated\\:PI-control\\:PI}{Control\\:PI}\\times\\:100\\%)\\)\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003eSerum Prostate-Specific Antigen (PSA) Estimation.\u003c/b\u003e Total serum PSA was estimated using PSA Enzyme Immunoassay test kit according to the manufacturer\u0026rsquo;s protocol. A microplate reader (iMark\u0026trade;, Bio-Rad Laboratories, USA) was used to read the optical density at 450nm within 15 minutes of colour development.\u003c/p\u003e\u003cp\u003e\u003cb\u003eHistopathological examination\u003c/b\u003e. Sections from prostate tissues were fixed in 10% neutral buffered formalin and processed for paraffin sections of 4 \u0026micro;m thickness. The sections were collected on glass slides, deparaffinized then stained with haematoxylin and eosin (H\u0026amp;E) for routine histopathological examination. Histopathological slides were analysed by an expert pathologist with knowledge in murine pathology, who was blind to the experimental conditions. A light microscope (Olympus CX23 Binocular Microscope) was used for image processing. The prostate tissue histology was evaluated by comparing the treated groups to those of the controls to reveal histological changes.\u003c/p\u003e\u003cdiv id=\"Sec7\" class=\"Section2\"\u003e\u003ch2\u003eStatistical Analysis\u003c/h2\u003e\u003cp\u003eAll data were expressed as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard error of mean (SEM), and comparison of mean parameters measured between experimental groups and control groups were determined using One-way ANOVA (Kruskal-Wallis test and Dunn's Multiple Comparison Test) using GraphPad Prism version 8.0.2. \u003cem\u003eP\u003c/em\u003e ˂ 0.05 was considered statistically significant.\u003c/p\u003e\u003c/div\u003e"},{"header":"Results","content":"\u003cp\u003eAndrogen signalling remains a major driver of prostate cell proliferation and BPH development; although serum testosterone levels decrease with advancing age (by about 2\u0026ndash;3% per year), the risk of developing BPH also increases with advancing age. This suggests that other factors enhance low serum testosterone to induce enlargement of the prostate, leading to BPH. In this study, we assessed the influence of four dietary components on the hormone-dependent enlargement of the prostate.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMinimum Testosterone Above Which BPH Induction Occurs\u003c/strong\u003e. To assess the potentiating effect of the dietary substances, the minimum amount of testosterone (\u003cstrong\u003eT)\u003c/strong\u003e above which significant enlargement of the prostate (BPH induction) occurred was determined. This was done by serially diluting the 400\u0026micro;g \u003cstrong\u003eT\u003c/strong\u003e into 200\u0026micro;g, 100\u0026micro;g, 50\u0026micro;g and 25\u0026micro;g and administering to obtain the minimum dose of 50\u0026micro;g \u003cstrong\u003eT\u003c/strong\u003e above which BPH induction occurred (Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003ea \u0026ndash; \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003ec). Subsequently, 100\u0026micro;g Testosterone was used as the positive control, while 50\u0026micro;g Testosterone was adopted as the minimum hormone to be administered concurrently with the dietary treatments to assess their potentiating effects.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eHigh Fat Diet (Pork and Beef sources) Increased Prostate Weight Index and Serum PSA.\u003c/strong\u003e Rats fed with a high-fat diet from pork and beef, respectively, with the minimum determined testosterone (50\u0026micro;g) dose showed significantly greater average PI of 2.56\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08g and 2.35\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06g compared with 1.30\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0 g for the 50\u0026micro;g treated only. Similarly, the serum PSA levels in the HFD groups were significantly higher compared with the 50\u0026micro;g treated only (Table \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e). Compared with the 50\u0026micro;g treated only, there were 96.3% and 80.2% increases in PI for pork and beef fat groups, respectively. The percentage increase in serum PSA for the pork and beef fat-fed rats respectively were 72.3% and 77.2% compared to the 50\u0026micro;g treated only. However, there was no significant difference in the PI and PSA levels induced by the pork fat-fed compared with the beef fat-fed groups.\u003c/p\u003e\n\u003cp\u003eTable 1: Percentage increase in PSA levels after Testosterone and dietary treatment\u003c/p\u003e\n\u003cp\u003e\u003cimg 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\"\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable \u003cspan class=\"InternalRef\"\u003e1\u0026nbsp;\u003c/span\u003e\u003c/strong\u003eHigh-fat diet enhanced low testosterone concentration to induce elevation of serum PSA. Data are expressed as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SEM (n\u0026thinsp;=\u0026thinsp;5). A one-way ANOVA was carried out to compare the minimum testosterone (50\u0026micro;g only) with the positive control (100\u0026micro;g T), 50\u0026micro;g \u003cstrong\u003eT\u003c/strong\u003e\u0026thinsp;+\u0026thinsp;Beef fat and 50\u0026micro;g \u003cstrong\u003eT\u003c/strong\u003e\u0026thinsp;+\u0026thinsp;Pork fat groups. * p-value\u0026thinsp;\u0026lt;\u0026thinsp;0.05.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eHigh Pork and Beef Fat Diet Promote Prostate Enlargement under Low Testosterone in Rats\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNext, we examined the effect of intake of high pork and high beef fat diets with the minimum \u003cstrong\u003eT\u003c/strong\u003e on the prostate histology by performing a histopathological examination. Changes in the prostate tissues were examined after H\u0026amp;E staining. The prostatic epithelial cells in the solvent control group were cuboidal and regular in size (Fig. \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e). Nevertheless, the epithelial cell layer in the positive control and the fat-fed rats were larger, and the epithelium became hyperplastic. Numerous papillae projecting into the lumen formed and further decreased the glandular luminal area (Fig. \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e). In the pork and beef fat diet groups, visible changes were observed showing much thickened epithelia and acini, hypertrophy, wider diameter of glandular lumen and hyperplasia with less marked inter-glandular stroma (Fig. \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e). There were no alterations in the histological appearance of the epithelial tissue linings in the solvent control. Thin layers of epithelia were observed, indicative of no induced hyperplasia.\u003c/p\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eHigh Salt and Sugar Diets slightly increased Prostate Index, serum PSA, and altered the Histology of Prostatic Tissues in BPH rats\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eIn comparison with the control group (minimum \u003cstrong\u003eT\u003c/strong\u003e only), rats on a high-salt diet plus minimum \u003cstrong\u003eT\u003c/strong\u003e showed no significant increase in PI and serum PSA levels, however, rats on the high-sugar diet plus 50\u0026micro;g\u003cstrong\u003eT\u003c/strong\u003e showed significantly higher PI (1.74\u0026thinsp;\u0026plusmn;\u0026thinsp;0.15) compared to the minimum \u003cstrong\u003eT\u003c/strong\u003e only control (1.30\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04). Similarly, the serum PSA levels (0.64\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01) was significantly higher compared with the 50\u0026micro;g\u003cstrong\u003eT\u003c/strong\u003e only treated (Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003eb). Significant increase in the levels of serum PSA was observed in high sugar treated group (0.64\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01 ng/ml) compared to the control (0.46\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01 ng/ml) (Table\u0026nbsp;3).\u003c/p\u003e\n\u003cp\u003eIn the HSu diet group, the increased PI and elevated serum PSA levels were accompanied by hyperplasia of glandular epithelium and inter-glandular stroma, which was absent in the control group, that showed thin glandular epithelia and fibro-muscular layers (Fig. \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e). However, the rats fed with HS displayed features similar to that of the 50\u0026micro;g \u003cstrong\u003eT\u003c/strong\u003e only control (Fig. \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n \u003ctable id=\"Tab2\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eSerum PSA levels after Testosterone and Dietary treatments.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eGroup (n\u0026thinsp;=\u0026thinsp;5)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003ePSA (ng/ml)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e% Increase\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eSolvent control\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.43\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e50\u0026micro;g \u003cem\u003eT\u003c/em\u003e only (minimum testosterone)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.46\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6.90\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e100\u0026micro;g \u003cem\u003eT\u003c/em\u003e only\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.75\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05**\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e74.42\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e50\u0026micro;g \u003cem\u003eT\u003c/em\u003e\u0026thinsp;+\u0026thinsp;High Salt\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.47\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e9.30\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e50\u0026micro;g \u003cem\u003eT\u003c/em\u003e\u0026thinsp;+\u0026thinsp;High Sugar\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.64\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e48.83\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable \u003cspan class=\"InternalRef\"\u003e2:\u0026nbsp;\u003c/span\u003e\u003c/strong\u003eHigh-sugar diet but not high-salt diet enhanced low testosterone concentration to induce the elevation of serum PSA. Data are expressed as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SEM (n\u0026thinsp;=\u0026thinsp;5). A one-way ANOVA was carried out to compare the minimum testosterone (50\u0026micro;g only) with the positive control (100\u0026micro;g T), 50\u0026micro;g \u003cstrong\u003eT\u003c/strong\u003e\u0026thinsp;+\u0026thinsp;High Sugar and 50\u0026micro;g \u003cstrong\u003eT\u003c/strong\u003e\u0026thinsp;+\u0026thinsp;High Salt groups. * p-value\u0026thinsp;\u0026lt;\u0026thinsp;0.05.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eHistopathological Examination\u003c/strong\u003e. Microscopic examination of H\u0026amp;E-stained sections of the ventral prostatic tissues taken from solvent control and the 50\u0026micro;g \u003cstrong\u003eT\u003c/strong\u003e only group showed normal structure in the form of numerous acini of different sizes and shapes containing homogenous acidophilic material. The lining epithelium of the acini was composed of two distinct cell types, namely, basal and acinar or principal cells. The acinar cells were columnar, extending from the basal lamina to the lumen of the duct (Fig. \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e). Conversely, prostates obtained from experimental groups C, D, E, F, and G rats displayed an obvious disruption of the histoarchitecture of the prostatic tissue shown as marked hyperplasia, irregular acinar folding with intraluminar projections, and congestion (Fig. \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e). However, similar to the prostate tissues from the positive control (C), glands with prominent widespread epithelial hyperplasia (j) and fibromuscular stromal hypertrophy (s) were identified in the experimental groups (Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e. D, E and G)\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eBPH is considered the most common urological disorder affecting men above 50 years of age with a prevalence of up to 85% in older age groups [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. It is characterized by non-cancerous enlargement and uncontrolled proliferation of smooth muscles and epithelial cells of the prostatic tissue. Clinically, BPH is manifested as LUTS with a significant negative impact on the quality of life [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. While earlier reports indicate that the development and progression of BPH are dependent on several factors [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e], there is not enough clarity on the role of dietary patterns in the development of this condition. In this study, we report that chronic intake of high-fat from red meat sources in the presence of low testosterone induces enlargement of the prostate in a rat model. Our data suggests that high-fat diet enhances low concentration of \u003cb\u003eT\u003c/b\u003e, which in itself, is not sufficient to induce prostate enlargement (hyperplasia) in the initiation of a BPH disease. Increase in PI and histological alterations in the prostate tissues have been important parameters in assessing prostate health and BPH development [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e, \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. HFD consumption has been strongly associated with higher BPH risk and potentially, cancer of the prostate (PCa) in several studies [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e, \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. How red meat fats enhanced low \u003cb\u003eT\u003c/b\u003e action to induce enlargement in this study is not fully understood, however, it has been reported by earlier studies that, HFD contain high amounts of saturated fatty acids (SFA) [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e, \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e] which plays a role in inflammation and enhances the production of reactive oxygen species (ROS) which worsens the oxidative stress symptoms in the prostate and as a consequence, hyperplasia is enhanced as a developmental adaptation under these conditions [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]. Another report consistent with ours was published by Shankar and colleagues, which indicated that an NF-κB reporter mice fed a high-fat diet (HFD) with a composition characteristic of a Western-style diet resulted in increased intra-prostatic inflammation and cell proliferation, leading to prostate gland enlargement [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e, \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. A recent report has indicated that HFD profoundly alters the expression of pro-proliferative genes and even an oncogene, such as MYC, resulting in the enhancement of MYC-driven transcriptional programs in prostate cells, leading to increased glandular cell proliferation and potential transformation of the prostate, in the case of cancer [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. These aforementioned reports support our findings that, even though excessive intake of animal fat is considered generally harmful to the body, it could also enhance prostate glandular cell proliferation under low \u003cb\u003eT\u003c/b\u003e levels in the development of enlarged prostate in men.\u003c/p\u003e\u003cp\u003eIn this study, we report that chronic consumption of high sugar enhanced low testosterone levels to induce enlargement of the prostate in rats. The reason for the observed synergy is not clearly decipherable. However, earlier studies have reported a strong association between fasting blood sugar and the size of the prostate [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]. Mechanistically, androgen treatment-induced prostatic hyperplasia \u003cem\u003ein vivo\u003c/em\u003e is dependent on increased expression of growth factors, mainly IGF-1, and its receptor IGF-1R, and increased expression of phosphorylated Akt as well as Bcl-2 [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. It also significantly decreases testosterone\u0026ndash;induced Bax / Bcl-2 ratio and P\u003csup\u003e21\u003c/sup\u003e activity, thereby inhibiting apoptosis [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. This may be one mechanism by which glucose enhances the action of low testosterone in promoting prostatic hyperplasia. Secondly, others have reported that high glucose treatment could induce proliferation and suppress apoptosis in prostatic epithelial cells through a different mechanism [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. Ye et al. (2011, 2018) found that under a high glucose environment, the level of reactive oxygen species (ROS) was significantly up-regulated, which induced cell proliferation and suppressed the apoptosis of prostate epithelial cells [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e, \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. Taken together, our findings suggest that high glucose treatment might facilitate BPH development, even in the presence of low testosterone.\u003c/p\u003e\u003cp\u003eAverage salt consumption in humans ranges from 9 to 12g per day (double the recommended dose), of which most of it is found in food condiments [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. It is known that high-salt (\u0026gt;\u0026thinsp;2g/day), which constitutes 40% of sodium intake, is a huge underlying risk factor for elevated arterial blood pressure, heart diseases, hypertension, and stroke [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e, \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. High serum sodium has been associated with the development of BPH and worse lower urinary tract symptoms [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e, \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. In our studies, a high salt diet over the period of 28 days did not enhance the minimum testosterone dose to increase the PI and therefore did not significantly differ when compared with that of NC. However, there were histological changes that appeared somewhat similar to the alterations observed in the positive control, as well as a slight increase in mean serum PSA, which could indicate an insidious inflammation flourishing in the prostate gland in response to the high salt intake. The mechanism by which high salt enhanced low \u003cb\u003eT\u003c/b\u003e action to cause the increased serum PSA of the HS rats' prostate is unclear. Salt is known to aggravate the symptoms of BPH and LUTS through the bladder and prostate innervation. In terms of mechanism, it is generally believed that it may be related to abnormal activation of the sympathetic nervous system [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e, \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. However, findings consistent with what we observed in this study was reported by Bello and colleagues (2023), who showed that low, standard, and high salt diets aggravated the pathology of testosterone-induced BPH in Wistar rats by promoting inflammation, oxidative stress, while suppressing apoptosis and angiogenesis [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e]. Although an insignificant enlargement of prostate glands was observed in the high salt group in this study, a significant increase in serum PSA levels was observed in the group compared with the controls. Therefore, the effect of high intake of salt observed in this study could be explained by pro-inflammatory activity of high dietary salt and its potential to enhance low \u003cb\u003eT\u003c/b\u003e to aggravate BPH pathophysiology by increasing epithelial thickness and polyps formation, which may have consequently raised the serum PSA levels.\u003c/p\u003e\u003cp\u003eTo the best of our knowledge, this study provides insight into the effect of dietary components and low testosterone on the risk of BPH in a rat model. The study showed the synergistic effect of high intake of red meat fat, table sugar, and table salt and their risk of BPH. However, there are some potential limitations of this study. Firstly, our study did not evaluate inflammatory cytokines in the prostate, as well as, gene expression profile of genes known to promote hyperplasia in the prostate, which would have provided more insight into the mechanism by which these dietary components enhance testosterone-mediated gene expression in the prostate.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThis study demonstrated that rat diets high in fat from red meat, high in sugar, and high in dietary salt promoted hyperplasia and enlargement of prostatic tissues under low testosterone levels. Therefore, it is concluded that dietary fats, sugars, and, to a lesser extent, salt intake enhanced the development of testosterone-induced BPH in rats. These observations suggest that dietary patterns are a contributing risk factor for BPH development. We recommend further molecular mechanistic studies to help clarify the causal role of various dietary patterns in the initiation or induction of BPH.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics, Consent to Participate, and Consent to Publish declarations.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics Statement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll experimental protocols and procedures were carried out in accordance with international ethical guidelines for the care and use of Laboratory Animals. Ethical approval was also obtained from the Animal Research Ethics Committee of the Kwame Nkrumah University of Science and Technology (KNUST/AREC/C.1- 0076).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publications\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable. Our manuscript contains data generated from original research involving animal model study; therefore, it does not contain any person’s data.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe datasets used and/or analyzed during the current study are available from the corresponding author upon reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no conflicts of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors’ contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eBS:\u003c/strong\u003e investigation, methodology, and writing original draft; \u003cstrong\u003eSA:\u003c/strong\u003e investigation, methodology; \u003cstrong\u003eKAAA\u003c/strong\u003e, \u003cstrong\u003eGKA:\u003c/strong\u003e supervision, review \u0026amp; editing; \u003cstrong\u003eEAN:\u003c/strong\u003e conceptualization, project administration, resources, supervision, investigation, methodology, formal analysis, writing, review \u0026amp; editing. All the authors read and approved the final manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgments\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors are very grateful to the Histopathologist, Mr. Clement Atobiga, and the entire staff at the Department of Pharmacology Animal House for their support in this study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor Information\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003csup\u003e1\u003c/sup\u003e Department of Physiology, School of Medical Sciences, Kwame Nkrumah University of Science and Technology, Kumasi\u003c/p\u003e\n\u003cp\u003e\u003csup\u003e2\u003c/sup\u003e Department of Pharmacology, Faculty of Pharmacy and Pharmaceutical Sciences, Kwame Nkrumah University of Science and Technology, Kumasi\u003c/p\u003e\n\u003cp\u003e\u003csup\u003e3\u0026nbsp;\u003c/sup\u003eDepartment of Surgery, School of Medicine and Dentistry, Kwame Nkrumah University of Science and Technology, Kumasi.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eLytton B, Emery JM, Harvard BM. 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J Clin Hypertens. 2019;21:1596\u0026ndash;601. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/jch.13664\u003c/span\u003e\u003cspan address=\"10.1111/jch.13664\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eBello II, Omigbodun A, Morhason-Bello I. Common salt aggravated pathology of testosterone-induced benign prostatic hyperplasia in adult male Wistar rat. BMC Urol. 2023;23:207. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1186/s12894-023-01371-x\u003c/span\u003e\u003cspan address=\"10.1186/s12894-023-01371-x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"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":"BPH, High fat diet (HFD), table sugar, salt, Prostatic Index (PI), Prostate Specific Antigen (PSA), hyperplasia, prostate enlargement","lastPublishedDoi":"10.21203/rs.3.rs-6946787/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6946787/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground: \u003c/strong\u003eBenign prostatic hyperplasia (BPH) is one of the most serious urinary system disorders in elderly men worldwide. It is characterized by prostate enlargement and bladder outlet obstruction and several symptoms clinically referred to as lower urinary tract symptoms. Reported risk factors include advancing age, circulatory androgens, and genetics. Nevertheless, the role of Dietary patterns in the development of this condition remains largely unresolved. This study, therefore, investigated the influence of consumption of high fat from beef and pork source, high table sugar and high salt on Effect of Dietary Patterns on testosterone-induced prostatic hyperplasia in a rat model.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethod\u003c/strong\u003e: Thirty (30) male Sprague Dawley (SD) rats weighing 200 - 250g were randomly assigned to the pork fat, beef fat, high sugar (HSu), and high salt (HS) groups. After establishing the minimum threshold amount of testosterone above which significant prostate enlargement occurred, 50µg, all experimental groups were administered with the minimum testosterone in addition to the various dietary treatments for four weeks. Positive and negative control groups were administered with 100µg and 50µg testosterone respectively, in addition to a standard rat diet for four weeks. After the 28 days, the final weights of animals were recorded, animals sacrificed, blood samples taken for serum PSA determination, prostates and seminal vesicles isolated, weighed, prostate weight index (PI) determined, and prostate tissues processed for histopathological analysis. Mean ± SD values were calculated for each group and compared for significant difference using ANOVA followed by post hoc test (Dunn's Multiple Comparison Test) at \u003cem\u003ep\u003c/em\u003e \u0026lt; 0.05.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults: \u003c/strong\u003eCompared to the negative control, the mean serum PSA levels and PI of the high-fat and HSu groups were significantly higher, but they did not differ from those of the positive control. Although the mean PSA of the HS group was significantly higher compared to the negative control, the mean PI did not differ from the negative control.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion: \u003c/strong\u003eCollectively, our findings indicate that consumption of high-fat from red meat and high sugar diets enhance the effects of low testosterone to induce prostate enlargement in a rat model.\u003c/p\u003e","manuscriptTitle":"Effect of Dietary Patterns on Testosterone-Induced Prostatic Hyperplasia in Rats","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-08-05 10:13:44","doi":"10.21203/rs.3.rs-6946787/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":"ff55d1dd-ccc6-48cc-9926-19f31503a0ee","owner":[],"postedDate":"August 5th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-09-03T19:53:49+00:00","versionOfRecord":[],"versionCreatedAt":"2025-08-05 10:13:44","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6946787","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6946787","identity":"rs-6946787","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}