Effect of age and gender on radioactive iodine induced apoptosis in rats kidney cells

Effect of age and gender on radioactive iodine induced apoptosis in rats kidney cells | 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 Article Effect of age and gender on radioactive iodine induced apoptosis in rats kidney cells Liang Yin, Yangyang Lin, Jianlan Yue, Zhinchun Lin, Hongtao Sun, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8555726/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: To explore the effects of age and gender on renal cell apoptosis induced by high-dose radioactive iodine ( 131 I) in rats. Methods: Forty-eight rats were divided into 4 groups (aged male group, aged female group, young male group, and young female group) according to age and sex, with 12 rats in each group, each group was randomly divided into control group and experimental group, with a total of 8 groups and 6 rats in each group. All rats in the experimental groups were intraperitoneally injected with 1.85 MBq/2g 131 I solution, and rats in control group were intraperitoneally injected with the same volume of 0.9% NaCl solution. All the rats were sacrificed after 4 weeks, the kidney sections were removed for immunohistochemical analysis of apoptosis-related proteins such as Bax, Bcl-2 and Caspase-3. Results: 4 weeks later, the expression levels of Caspase-3 and Bax proteins in all the experimental groups were significantly increased, while the expression level of Bcl-2 protein and Bcl-2/Bax ratio were significantly decreased. Compared with the aged rats, the expression level of apoptosis in young rats was increased more significantly, and the difference was statistically significant. There was no significant difference in the apoptosis level between male and female rats. Conclusion: A single high-dose of 131 I can lead to a significant increase in the level of renal cell apoptosis in rats, and the level of apoptosis is more obvious in young rats than aged rats, but the gender of the rats is not related to the level of apoptosis. Biological sciences/Biochemistry Health sciences/Nephrology Biological sciences/Physiology radioactive iodine age gender renal cell apoptosis rats Figures Figure 1 Figure 2 Figure 3 Introduction Kidney is one of the most sensitive organs to radiation injury in the abdominal organs [1] , and it is also one of the dose-limiting organs in radionuclide therapy [2,3] . Studies have shown that the pathological changes of kidney injury induced by radiation are mainly manifested as tubular and glomerular atrophy and interstitial fibrosis [4] . Ionizing radiation can trigger the continuous release of Reactive oxygen species (ROS), which leads to the activation of local inflammatory mediators and the occurrence of acute and chronic oxidative stress, resulting in severe lipid peroxidation damage in kidney tissue, thereby causing cell apoptosis [5] . High-dose 131 I therapy is one of the main treatment methods for differentiated thyroid carcinoma, a common endocrine tumor. Since 131 I is mainly excreted by the kidneys, this excretion pathway may expose 131 I the relevant urinary system organs to radiation [6] . In addition, sodium iodide symporter (NIS) protein is present in renal tubular epithelial cells that can actively take up 131 I [7] . Therefore, there is a theoretical risk of radiation-induced renal cell apoptosis in high-dose 131 I treatment. Age and gender can significantly affect the risk spectrum of radiation damage by affecting tissue sensitivity, repair ability and hormone levels. At present, the studies on the effects of age and gender on individual radiation damage are mostly focused on the field of external irradiation, and the research data on the effects of age and gender on radionuclide internal radiation therapy nephrotoxicity are few. The aim of the present study is to evaluate the effects of age and gender on renal cell apoptosis induced by high-dose 131 I in rats. 1 Materials and methods 1.1 Experimental Animals In the present study, 48 healthy Wistar albino rats, respectively for 6 weeks of young male group and young female group rats each 12, 30 weeks aged male group and aged female group rats each 12. They were housed in cages, 2 ~ 3 per cage, in the windowless animal quarter automatic temperature (22 ± 1℃) and lighting controls (12 h light/12 h dark cycle) were performed, relative humidity ranged from 55% to 60%, and supplied with standard food and water ad libitum. All experimental procedures were carried out according to the relevant experimental guidelines and in accordance with the regulations of the National Animal Protection Association. The animal experiment study protocol was approved by the Animal Management and Use Committee of the National Cancer Center. The euthanasia method was selected for terminal procedures requiring tissue collection. Throughout the 28-day study period, animals were monitored twice daily for clinical signs, behavior, and overall health. Humane endpoints were applied to minimize pain and distress. All procedures were conducted by trained personnel, and no unexpected deaths occurred. The study has been approved by the ethics committee of Tianjin Medical University General Hospital (Protocol Number: 2021-053) and adhered to the ARRIVE (Animal Research: Reporting In Vivo Experiments) guidelines. 1.2 Grouping and intervention of experimental animals Twelve rats in each group (aged male group, aged female group, young male group, and young female group)were randomly divided into two groups as follows: the control group (n = 6) and the experimental group (n = 6). The rats in the experimental group were intraperitoneally injected with 131 I solution at a dose of 0.05mCi/2g, and the rats in the control group were intraperitoneally injected with the same volume of 0.9% NaCl solution. All control groups and experimental groups rats were euthanized after 4 weeks and immediately after the rats were euthanized, right nephrectomy was performed. (Euthanasia method: Rats were first placed in an induction chamber prefilled with a high concentration of isoflurane in medical air, and depth of anesthesia was confirmed by loss of the pedal withdrawal reflex before any procedure. Following induction, rats were rapidly euthanized by cervical dislocation while under deep anesthesia). 1.3 Kidney immunohistochemistry The rat kidneys were fixed in buffered neutral 10% formaldehyde for 24 hours. The tissues were cut sagittally and half of each kidney was sampled and they were subjected to alcohol processing for 12 hours. Five micrometer thick sections were obtained from the paraffin embedded tissues. Interpretation of immunohistochemical results: cysteinyl aspartate specific proteinase-3(Caspase-3), B-cell lymphoma-2(Bcl-2) and Bcl-2 assaciated X Protein(Bax) were selected for analysis, and the brown yellow was positive expression staining of apoptotic tissue. The long and short axes of mouse kidney tissue sections were determined by vernier caliper, and the lines were marked on the back. A relatively complete cell field was taken at the center site and the five vertices of the pentagon, respectively. Then Image-Pro Plus 6.0 professional Image analysis software was used to measure the area and integrated optical density (IOD), and the average optical density was calculated. The higher the average optical density value, the higher the expression level of the index it represents. 1.4 Statistical Analysis SPSS 22.0 statistical software was used for statistical analysis. The results are expressed as mean ± standard deviation. The average optical density of Caspase-3, Bcl-2 and Bax and Bcl-2/Bax ratio were compared between groups by two independent samples t test . p values below 0.05 were considered as statistically significant. 2 Results 2.1 The aged male group and the aged female group were combined together as the aged group, and the young male group and the young female group were combined as the young group. The apoptosis data of kidney cells in different age groups were compared (Table 1) : 2.1.1 Aged control group with young control group rats Caspase-3, the Bcl-2, Bax and Bcl-2/Bax values were no statistical difference ( P > 0.05). 2.1.2 After 4 weeks of high-dose 131 I administration, the levels of apoptosis in the aged experimental group and the young experimental group were significantly increased, the levels of Caspase-3 and Bax were significantly higher than those in the control group(Figure 1), and the Bcl-2 and Bcl-2/Bax ratio were significantly lower than those in the control group ( P<0. 05). 2.1.3 After 4 weeks of high-dose 131 I administration, the apoptosis level of the young experimental group was higher than that of the aged experimental group, the Caspase-3 and Bax levels of the young experimental group were higher than those of the aged experimental group, and the Bcl-2 and Bcl-2/Bax value were lower than those of the aged experimental group, and the differences were statistically significant ( P<0. 05). 2.2 The aged male group and the young male group were combined into the male group, and the aged female group and the young female group were combined into the female group. The renal cell apoptosis data of rats in different gender groups were compared (Table 2) : 2.2.1 There were no significant differences in Caspase-3, Bcl-2, Bax and Bcl-2/Bax ratio between male control group and female control group ( all P > 0.05). 2.2.2 After 4 weeks of high-dose 131 I administration, the levels of apoptosis in the male group and female group were increased significantly, the Caspase 3, Bax levels higher than the control group obviously(Figure 2), the Bcl-2 and the Bcl-2/Bax value significantly lower than control group(Figure 3), difference has statistical significance (P < 0.05). 2.2.3 After 4 weeks of high-dose 131 I administration, there was no significant difference in Caspase-3, Bax, Bcl-2 and Bcl-2/Bax ratio between male experimental group and female experimental group ( all P > 0.05). 3 Discussion The results of the study showed that high-dose 131 I could enhance the expression of apoptosis in renal cells of rats, which was manifested as the up-regulation of Caspase-3 and Bax protein expression, and the down-regulation of Bcl-2 protein expression and Bcl-2/Bax ratio, suggesting that the activation of apoptotic pathway may be the cause of renal dysfunction induced by 131 I. Compared with the aged rats, the expression level of apoptosis in the young rats was relatively more obvious, and the difference was statistically significant. However, there was no significant difference in apoptosis-related immunohistochemical expression levels between male and female rats. Apoptosis, also known as programmed cell death (PCD), is characterized by cell shrinkage, membrane bleb, nuclear chromatin condensation and nuclear fragmentation [8] . Caspase-3, a biomarker of apoptosis, is involved in the final stage of apoptosis [9] . Studies have shown that radiation induces renal tissue damage and induces apoptosis by producing ROC and increasing the expression of caspase-3 in renal tubular cells [10] . Bcl-2 is an anti-apoptotic protein, which can inhibit cell apoptosis caused by a variety of injuries in cells, thereby enabling cell survival. When the expression level of Bcl-2 is increased, it can significantly inhibit the speed of cell apoptosis, thereby slowing down the occurrence of cell apoptosis [11-14] . The pro-apoptotic effector protein Bax is the homologous gene of Bcl-2 and has the biological effect of antagonizing Bcl-2. Bax protein can induce cells to go to apoptosis [15, 16] . The results of the present study showed that the expression levels of apoptotic proteins in the renal tissue of rats were increased to varying degrees after high-dose 131 I administration. These observations are basically consistent with previous reports, suggesting that the apoptotic expression of animal kidney cells will be enhanced after radiation exposure [17] . Regarding the effect of age on radiosensitivity. Experimental studies on animal external irradiation have confirmed that young animals are more susceptible to the effects of ionizing radiation than adult animals [18-24] , that is, young animals are relatively more sensitive to radiation. Clinical studies have also found that children often have higher radiation sensitivity in the process of radiotherapy [25] . However, Saracyn et al suggested that age had no significant effect on renal function during PRRT, which was different from external beam radiotherapy [26] . The results of the study showed that after the rats were given a high-dose of 131 I, the apoptosis level of the aged group and the young group increased, and the apoptosis of the kidney cells in the young rats was more obvious than that in the aged rats, indicating that the kidney cells in the young rats were more radiosensitive. In terms of the effect of gender on radiosensitivity, animal experiments with external irradiation have shown that there may be significant differences in the sensitivity of male and female to ionizing radiation [27, 28] . One of the possible reasons for this difference is the difference in sex hormones [29] . Recent radiogenomic studies have also shown that the ability to repair radiation-induced DNA breaks is significantly different between males and females [30] . In an analysis of 807 patients with neuroendocrine tumors treated with PRRT ( 177 Lu or 90 Y or 177 a combination of Lu and 90 Y), Bodei et al found a higher incidence of nephrotoxicity in male patients than in female patients [31] . However, there are also some studies that analyzed the side effects of PRRT according to gender, showing no difference in nephrotoxicity between male and female [26,32] . In the study, there was no gender difference in the radiosensitivity of rats to renal cell apoptosis after high dose 131 I administration. The reason may be related to the difference between internal irradiation and external irradiation, and the small sample size and short study time. In general, there are few literatures on the effect of gender and age on renal damage caused by radionuclide internal radiation therapy, and no unified opinion has been reached, and more studies are needed to confirm. Declarations Conflict of Interest: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. Informed consent: The animal experimentation has been approved by the ethics committee of Tianjin Medical University General Hospital. Funding: No Funding. Author Contribution Liang Yin and Yangyang Lin wrote the main manuscript text. Jianlan Yue and Zhinchun Lin prepared figures. All authors reviewed the manuscript. Acknowledgement We are indebted to Weilong Li, Xiaolan Lv, and the technical staff of the Division of Nuclear Medicine of Tianjin Medical University General Hospital for their technical assistance and support. Data Availability The datasets used and/or analysed during the current study available from the corresponding author on reasonable request. References Fuma S, Kubota Y, Ihara S, et al. Radiocaesium contamination of wild boars in Fukushima and surrounding regions after the Fukushima nuclear accident [J]. J Environ Radioact, 2016, 164: 60-64. Stolniceanu CR, Nistor I, Bilha SC, et al. Nephrotoxicity/renal failure after therapy with 90Yttrium- and 177Lutetium-radiolabeled somatostatin analogs in different types of neuroendocrine tumors: a systematic review [J]. Nucl Med Commun, 2020, 41(7): 601-617. Parihar AS, Chopra S, Prasad V. Nephrotoxicity after radionuclide therapies [J]. Transl Oncol, 2022, 15(1): 101295. 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Effects of irradiation on cumulative mortality in rats: shifting toward a younger age of death [J]. J Radiat Res. 2023, 64(2): 412-419. Milano MT, Marks LB, Olch AJ, et al. Comparison of Risks of Late Effects From Radiation Therapy in Children Versus Adults: Insights From the QUANTEC, HyTEC, and PENTEC Efforts [J]. Int J Radiat Oncol Biol Phys, 2024, 119(2): 387-400. Saracyn M, Durma AD, Bober B, et al. Long-Term Complications of Radioligand Therapy with Lutetium-177 and Yttrium-90 in Patients with Neuroendocrine Neoplasms [J]. Nutrients, 2022, 15(1): 185. Varma C, Schroeder MK, Price BR, et al. Long-Term, Sex-Specific Effects of GCRsim and Gamma Irradiation on the Brains, Hearts, and Kidneys of rats with Alzheimer's Disease Mutations [J]. Int J Mol Sci, 2024, 25(16): 8948. Holmes-Hampton GP, Soni DK, Kumar VP, et al. Time- and sex-dependent delayed effects of acute radiation exposure manifest via miRNA dysregulation [J]. iScience, 2024, 27(2): 108867. 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Tables Table 1: Mean optical density values of Caspase-3, Bcl-2 and Bax and Bcl-2/Bax ratio in kidney of rats of different age groups ( x̄ ±s) Grouping Caspase-3 Bax Bcl-2 Bcl-2/Bax aged group Control group 21.18±2.27 16.25±2.34 31.46±5.55 1.97±0.37 Experimental group 27.82±5.78 20.78±3.66 26.24±6.18 1.27±0.25 Young group Control group 21.96±4.97 17.03±4.07 32.56±7.52 2.05±0.81 Experimental group 34.96±8.49 24.67±3.75 20.87±5.68 0.87±0.28 The expression levels of Caspase-3, Bax, Bcl-2 and Bcl-2/Bax were significantly different between the control group and the experimental group (t =3.705, 3.615, -2.178, -5.373, respectively ; P =0.001, 0.002, 0.040, 0.000, respectively). There were significant differences in Caspase-3, Bax, Bcl-2 and Bcl-2/Bax between the control group and the experimental group (t =4.579, 4.782, -4.297, -4.813, respectively; all P =0.000). There were no significant differences in Caspase-3, Bax, Bcl-2 and Bcl-2/Bax ratio between the aged control group and the young control group (t =-0.492, -0.580, -0.404, -0.347, respectively ; P =0.627, 0.568, 0.690, 0.732, respectively). There were significant differences in Caspase-3, Bax, Bcl-2 and Bcl-2/Bax ratio between the aged experimental group and the young experimental group (t =-2.406, -2.567, 2.217, 3.749, respectively ; P =0.025, 0.018, 0.037, 0.001, respectively). Table 2: Mean optical density values of Caspase-3, Bcl-2 and Bax and Bcl-2/Bax ratio in kidney of rats of different gender groups ( x̄ ±s ) Grouping Caspase-3 Bax Bcl-2 Bcl-2/Bax Male group Control group 21.66±3.98 16.71±3.62 32.44±6.33 2.05±0.69 Experimental group 31.73±7.41 22.88±3.43 23.96±6.45 1.07±0.32 Female group Control group 21.47±3.79 16.57±3.05 31.59±6.89 1.97±0.56 Experimental group 31.06±8.83 22.57±4.89 23.15±6.65 1.07±0.35 There were significant differences in Caspase-3, Bax, Bcl-2 and Bcl-2/Bax ratio between the control group and the experimental group in male rats (t =4.143, 4.294, -3.252, -4.457, respectively ; P =0.000, 0.000, 0.004, 0.000, respectively). There were significant differences in Caspase-3, Bax, Bcl-2 and Bcl-2/Bax between the control group and the experimental group in female rats (t =3.454, 3.607, -3.047, -4.712; P =0.002, 0.002, 0.006, 0.000, respectively ). There were no significant differences in Caspase-3, Bax, Bcl-2 and Bcl-2/Bax between male control group and female control group (t =0.120, 0.098, 0.319, 0.325, respectively ; P =0.906, 0.923, 0.753, 0.749, respectively). There were no significant differences in Caspase-3, Bax, Bcl-2 and Bcl-2/Bax between male and female rats (t =0.202, 0.181, 0.302, -0.013, respectively ; P =0.842, 0.858, 0.765, 0.990, respectively). 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. We do this by developing innovative software and high quality services for the global research community. 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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-8555726","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":589747386,"identity":"5160ddf6-883d-45a0-a25e-65141a071bd9","order_by":0,"name":"Liang Yin","email":"","orcid":"","institution":"Pingjin Hospital, Characteristic Medical Center of Chinese People’s Armed Police Forces","correspondingAuthor":false,"prefix":"","firstName":"Liang","middleName":"","lastName":"Yin","suffix":""},{"id":589747387,"identity":"af874378-e052-4798-8156-dad827b864fb","order_by":1,"name":"Yangyang Lin","email":"","orcid":"","institution":"Tianjin Children’ s Hospital","correspondingAuthor":false,"prefix":"","firstName":"Yangyang","middleName":"","lastName":"Lin","suffix":""},{"id":589747388,"identity":"cb358346-02f5-4745-865b-45f2d510cc87","order_by":2,"name":"Jianlan Yue","email":"","orcid":"","institution":"Pingjin Hospital, Characteristic Medical Center of Chinese People’s Armed Police Forces","correspondingAuthor":false,"prefix":"","firstName":"Jianlan","middleName":"","lastName":"Yue","suffix":""},{"id":589747389,"identity":"f550ae55-b87b-4777-a1fd-0826eb92804b","order_by":3,"name":"Zhinchun Lin","email":"","orcid":"","institution":"Pingjin Hospital, Characteristic Medical Center of Chinese People’s Armed Police Forces","correspondingAuthor":false,"prefix":"","firstName":"Zhinchun","middleName":"","lastName":"Lin","suffix":""},{"id":589747390,"identity":"69747eee-520b-40af-8cc9-937a491bdaf8","order_by":4,"name":"Hongtao Sun","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAy0lEQVRIiWNgGAWjYBACeYaDjQ8SeNjs7NsbiNRi2Hj4sMEHGb5kA54DxFpz+Fia5AwbOcYNEglE6mBsO2MgzZNjxmwu+XjjDYYam2iCWth5zhgY85xJ47OcnVZswXAsLbeBoC0zzhgk8/YcY2a4nWMmwdhwmLAWhvtvDA7z/vvP2HDzDLFaDhxLbJzBw8a44QYPkVoMGw4fZvjAw5Ys2QP0SwIxfgFGZfsPUFTysx/eeONDjQ0RDkMCBkRHDZIWUnWMglEwCkbByAAA6FRCWvCnpqQAAAAASUVORK5CYII=","orcid":"","institution":"Pingjin Hospital, Characteristic Medical Center of Chinese People’s Armed Police Forces","correspondingAuthor":true,"prefix":"","firstName":"Hongtao","middleName":"","lastName":"Sun","suffix":""},{"id":589747391,"identity":"a7a2d975-a143-4c48-82db-13c677f96428","order_by":5,"name":"Zhaowei Meng","email":"","orcid":"","institution":"Tianjin Medical University General Hospital","correspondingAuthor":false,"prefix":"","firstName":"Zhaowei","middleName":"","lastName":"Meng","suffix":""}],"badges":[],"createdAt":"2026-01-09 01:38:17","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-8555726/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8555726/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":102827762,"identity":"adfa5fc9-dafb-4b4a-8e5e-dc939fb9efc2","added_by":"auto","created_at":"2026-02-17 09:12:15","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":1232267,"visible":true,"origin":"","legend":"\u003cp\u003eCaspase-3 staining of aged male group (×400) : A: Control group sections show normal tubular epithelial cell; B: Experimental group sections show apoptotic epithelial tubule cells.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-8555726/v1/09459bcf8cb152da78a867c8.png"},{"id":102827735,"identity":"080213a5-03ae-4d6e-9e90-a419cb225a3d","added_by":"auto","created_at":"2026-02-17 09:12:12","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":1349456,"visible":true,"origin":"","legend":"\u003cp\u003eBax staining of young male group (×400) : C: Control group sections show normal tubular epithelial cell; D: Experimental group sections show apoptotic epithelial tubule cells.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-8555726/v1/0257acb54219cbb8f6da2e65.png"},{"id":102827736,"identity":"e66e4fed-0d4d-4dcb-92d7-be7c5be846fa","added_by":"auto","created_at":"2026-02-17 09:12:12","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":1194679,"visible":true,"origin":"","legend":"\u003cp\u003eBcl-2 staining of aged female group (×400) : E: Control group sections show normal tubular epithelial cell; F: Experimental group sections show apoptotic epithelial tubule cells.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-8555726/v1/493faf7234996f8eac8b77b2.png"},{"id":104401720,"identity":"7cde8a33-e3fb-4798-9f37-873fbdd5cec7","added_by":"auto","created_at":"2026-03-11 12:13:21","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":5830750,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8555726/v1/bc6b29a2-3c22-4cfa-96d3-b7a24c486767.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Effect of age and gender on radioactive iodine induced apoptosis in rats kidney cells","fulltext":[{"header":"Introduction","content":"\u003cp\u003eKidney is one of the most sensitive organs to radiation injury in the abdominal organs\u003csup\u003e[1]\u003c/sup\u003e, and it is also one of the dose-limiting organs in radionuclide therapy\u003csup\u003e[2,3]\u003c/sup\u003e. Studies have shown that the pathological changes of kidney injury induced by radiation are mainly manifested as tubular and glomerular atrophy and interstitial fibrosis\u003csup\u003e[4]\u003c/sup\u003e. Ionizing radiation can trigger the continuous release of Reactive oxygen species (ROS), which leads to the activation of local inflammatory mediators and the occurrence of acute and chronic oxidative stress, resulting in severe lipid peroxidation damage in kidney tissue, thereby causing cell apoptosis\u003csup\u003e[5]\u003c/sup\u003e. High-dose \u003csup\u003e131\u003c/sup\u003eI therapy is one of the main treatment methods for differentiated thyroid carcinoma, a common endocrine tumor. Since \u003csup\u003e131\u003c/sup\u003eI is mainly excreted by the kidneys, this excretion pathway may expose \u003csup\u003e131\u003c/sup\u003eI the relevant urinary system organs to radiation\u003csup\u003e[6]\u003c/sup\u003e. In addition, sodium iodide symporter (NIS) protein is present in renal tubular epithelial cells that can actively take up \u003csup\u003e131\u003c/sup\u003eI\u003csup\u003e[7]\u003c/sup\u003e. Therefore, there is a theoretical risk of radiation-induced renal cell apoptosis in high-dose \u003csup\u003e131\u003c/sup\u003eI treatment.\u003c/p\u003e \u003cp\u003eAge and gender can significantly affect the risk spectrum of radiation damage by affecting tissue sensitivity, repair ability and hormone levels. At present, the studies on the effects of age and gender on individual radiation damage are mostly focused on the field of external irradiation, and the research data on the effects of age and gender on radionuclide internal radiation therapy nephrotoxicity are few. The aim of the present study is to evaluate the effects of age and gender on renal cell apoptosis induced by high-dose \u003csup\u003e131\u003c/sup\u003eI in rats.\u003c/p\u003e"},{"header":"1 Materials and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e1.1 Experimental Animals\u003c/h2\u003e \u003cp\u003eIn the present study, 48 healthy Wistar albino rats, respectively for 6 weeks of young male group and young female group rats each 12, 30 weeks aged male group and aged female group rats each 12. They were housed in cages, 2\u0026thinsp;~\u0026thinsp;3 per cage, in the windowless animal quarter automatic temperature (22\u0026thinsp;\u0026plusmn;\u0026thinsp;1℃) and lighting controls (12 h light/12 h dark cycle) were performed, relative humidity ranged from 55% to 60%, and supplied with standard food and water ad libitum. All experimental procedures were carried out according to the relevant experimental guidelines and in accordance with the regulations of the National Animal Protection Association. The animal experiment study protocol was approved by the Animal Management and Use Committee of the National Cancer Center. The euthanasia method was selected for terminal procedures requiring tissue collection. Throughout the 28-day study period, animals were monitored twice daily for clinical signs, behavior, and overall health. Humane endpoints were applied to minimize pain and distress. All procedures were conducted by trained personnel, and no unexpected deaths occurred. The study has been approved by the ethics committee of Tianjin Medical University General Hospital (Protocol Number: 2021-053) and adhered to the ARRIVE (Animal Research: Reporting In Vivo Experiments) guidelines.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e1.2 Grouping and intervention of experimental animals\u003c/h2\u003e \u003cp\u003eTwelve rats in each group (aged male group, aged female group, young male group, and young female group)were randomly divided into two groups as follows: the control group (n\u0026thinsp;=\u0026thinsp;6) and the experimental group (n\u0026thinsp;=\u0026thinsp;6). The rats in the experimental group were intraperitoneally injected with \u003csup\u003e131\u003c/sup\u003eI solution at a dose of 0.05mCi/2g, and the rats in the control group were intraperitoneally injected with the same volume of 0.9% NaCl solution.\u003c/p\u003e \u003cp\u003eAll control groups and experimental groups rats were euthanized after 4 weeks and immediately after the rats were euthanized, right nephrectomy was performed. (Euthanasia method: Rats were first placed in an induction chamber prefilled with a high concentration of isoflurane in medical air, and depth of anesthesia was confirmed by loss of the pedal withdrawal reflex before any procedure. Following induction, rats were rapidly euthanized by cervical dislocation while under deep anesthesia).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e1.3 Kidney immunohistochemistry\u003c/h2\u003e \u003cp\u003eThe rat kidneys were fixed in buffered neutral 10% formaldehyde for 24 hours. The tissues were cut sagittally and half of each kidney was sampled and they were subjected to alcohol processing for 12 hours. Five micrometer thick sections were obtained from the paraffin embedded tissues.\u003c/p\u003e \u003cp\u003eInterpretation of immunohistochemical results: cysteinyl aspartate specific proteinase-3(Caspase-3), B-cell lymphoma-2(Bcl-2) and Bcl-2 assaciated X Protein(Bax) were selected for analysis, and the brown yellow was positive expression staining of apoptotic tissue. The long and short axes of mouse kidney tissue sections were determined by vernier caliper, and the lines were marked on the back. A relatively complete cell field was taken at the center site and the five vertices of the pentagon, respectively. Then Image-Pro Plus 6.0 professional Image analysis software was used to measure the area and integrated optical density (IOD), and the average optical density was calculated. The higher the average optical density value, the higher the expression level of the index it represents.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e1.4 Statistical Analysis\u003c/h2\u003e \u003cp\u003eSPSS 22.0 statistical software was used for statistical analysis. The results are expressed as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation. The average optical density of Caspase-3, Bcl-2 and Bax and Bcl-2/Bax ratio were compared between groups by two independent samples t \u003cem\u003etest\u003c/em\u003e. p values below 0.05 were considered as statistically significant.\u003c/p\u003e \u003c/div\u003e"},{"header":"2 Results","content":"\u003cp\u003e2.1\u0026nbsp;The aged male group and the aged female group were combined together as the aged group, and the young male group and the young female group were combined as the young group. The apoptosis data of kidney cells in different age groups were compared (Table 1) :\u003c/p\u003e\n\u003cp\u003e2.1.1 Aged control group with young control group rats Caspase-3, the Bcl-2, Bax and Bcl-2/Bax values were no statistical difference (\u003cem\u003eP\u003c/em\u003e \u0026gt; 0.05).\u003c/p\u003e\n\u003cp\u003e2.1.2 After 4 weeks of high-dose \u003csup\u003e131\u003c/sup\u003eI administration, the levels of apoptosis in the aged experimental group and the young experimental group were significantly increased, the levels of Caspase-3 and Bax were significantly higher than those in the control group(Figure 1), and the Bcl-2 and Bcl-2/Bax ratio were significantly lower than those in the control group (\u003cem\u003eP\u0026lt;0.\u003c/em\u003e05).\u003c/p\u003e\n\u003cp\u003e2.1.3 After 4 weeks of high-dose \u003csup\u003e131\u003c/sup\u003eI administration, the apoptosis level of the young experimental group was higher than that of the aged experimental group, the Caspase-3 and Bax levels of the young experimental group were higher than those of the aged experimental group, and the Bcl-2 and Bcl-2/Bax value were lower than those of the aged experimental group, and the differences were statistically significant (\u003cem\u003eP\u0026lt;0.\u003c/em\u003e05).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e2.2\u0026nbsp;The aged male group and the young male group were combined into the male group, and the aged female group and the young female group were combined into the female group. The renal cell apoptosis data of rats in different gender groups were compared (Table 2) :\u003c/p\u003e\n\u003cp\u003e2.2.1 There were no significant differences in Caspase-3, Bcl-2, Bax and Bcl-2/Bax ratio between male control group and female control group (\u003cem\u003eall P\u003c/em\u003e \u0026gt; 0.05).\u003c/p\u003e\n\u003cp\u003e2.2.2 After 4 weeks of high-dose \u003csup\u003e131\u003c/sup\u003eI administration, the levels of apoptosis in the male group and female group were increased significantly, the Caspase 3, Bax levels higher than the control group obviously(Figure 2), the Bcl-2 and the Bcl-2/Bax value significantly lower than control group(Figure 3), difference has statistical significance (P \u0026lt; 0.05).\u003c/p\u003e\n\u003cp\u003e2.2.3 After 4 weeks of high-dose \u003csup\u003e131\u003c/sup\u003eI administration, there was no significant difference in Caspase-3, Bax, Bcl-2 and Bcl-2/Bax ratio between male experimental group and female experimental group (\u003cem\u003eall P\u003c/em\u003e \u0026gt; 0.05).\u003c/p\u003e"},{"header":"3 Discussion","content":"\u003cp\u003eThe results of the study showed that high-dose \u003csup\u003e131\u003c/sup\u003eI could enhance the expression of apoptosis in renal cells of rats, which was manifested as the up-regulation of Caspase-3 and Bax protein expression, and the down-regulation of Bcl-2 protein expression and Bcl-2/Bax ratio, suggesting that the activation of apoptotic pathway may be the cause of renal dysfunction induced by \u003csup\u003e131\u003c/sup\u003eI. Compared with the aged rats, the expression level of apoptosis in the young rats was relatively more obvious, and the difference was statistically significant. However, there was no significant difference in apoptosis-related immunohistochemical expression levels between male and female rats.\u003c/p\u003e\n\u003cp\u003eApoptosis, also known as programmed cell death (PCD), is characterized by cell shrinkage, membrane bleb, nuclear chromatin condensation and nuclear fragmentation\u003csup\u003e[8]\u003c/sup\u003e. Caspase-3, a biomarker of apoptosis, is involved in the final stage of apoptosis\u003csup\u003e[9]\u003c/sup\u003e. Studies have shown that radiation induces renal tissue damage and induces apoptosis by producing ROC and increasing the expression of caspase-3 in renal tubular cells\u003csup\u003e[10]\u003c/sup\u003e. Bcl-2 is an anti-apoptotic protein, which can inhibit cell apoptosis caused by a variety of injuries in cells, thereby enabling cell survival. When the expression level of Bcl-2 is increased, it can significantly inhibit the speed of cell apoptosis, thereby slowing down the occurrence of cell apoptosis\u003csup\u003e[11-14]\u003c/sup\u003e. The pro-apoptotic effector protein Bax is the homologous gene of Bcl-2 and has the biological effect of antagonizing Bcl-2. Bax protein can induce cells to go to apoptosis\u003csup\u003e[15, 16]\u003c/sup\u003e. The results of the present study showed that the expression levels of apoptotic proteins in the renal tissue of rats were increased to varying degrees after high-dose \u003csup\u003e131\u003c/sup\u003eI administration. These observations are basically consistent with previous reports, suggesting that the apoptotic expression of animal kidney cells will be enhanced after radiation exposure\u003csup\u003e[17]\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003eRegarding the effect of age on radiosensitivity. Experimental studies on animal external irradiation\u0026nbsp;have confirmed that young animals are more susceptible to the effects of ionizing radiation than adult animals\u003csup\u003e[18-24]\u003c/sup\u003e, that is, young animals are relatively more sensitive to radiation. Clinical studies have also found that children often have higher radiation sensitivity\u0026nbsp;in the process of radiotherapy\u003csup\u003e[25]\u003c/sup\u003e. However, Saracyn et al suggested that age had no significant effect on renal function during PRRT, which was different from external beam radiotherapy\u003csup\u003e[26]\u003c/sup\u003e.\u0026nbsp;The results of the\u0026nbsp;study showed that after the\u0026nbsp;rats\u0026nbsp;were given a high-dose of \u003csup\u003e131\u003c/sup\u003eI, the apoptosis level of the aged group and the young group increased, and the apoptosis of the kidney cells in the young rats was more obvious than that in the aged rats, indicating that the kidney cells in the young rats were more radiosensitive.\u003c/p\u003e\n\u003cp\u003eIn terms of the effect of gender on radiosensitivity, animal experiments with external irradiation have shown that there may be significant differences\u0026nbsp;in the sensitivity of male and female to ionizing radiation\u003csup\u003e[27, 28]\u003c/sup\u003e. One of the possible reasons for this difference is the difference in sex hormones\u003csup\u003e[29]\u003c/sup\u003e. Recent radiogenomic studies have also shown that the ability to repair radiation-induced DNA breaks is significantly different between males and females\u003csup\u003e[30]\u003c/sup\u003e. In an analysis of 807 patients with neuroendocrine tumors treated with PRRT (\u003csup\u003e177\u003c/sup\u003eLu or\u003csup\u003e90\u003c/sup\u003e Y or\u003csup\u003e177\u003c/sup\u003e a combination of Lu and\u003csup\u003e90\u003c/sup\u003e Y), Bodei et al found a higher incidence of nephrotoxicity in male patients than in female patients\u003csup\u003e[31]\u003c/sup\u003e. However, there are also some studies that analyzed the side effects of PRRT according to gender, showing no difference in nephrotoxicity between male and female\u003csup\u003e[26,32]\u003c/sup\u003e. In the\u0026nbsp;study, there was no gender difference in the radiosensitivity of\u0026nbsp;rats\u0026nbsp;to renal cell apoptosis after high dose \u003csup\u003e131\u003c/sup\u003eI administration. The reason may be related to the difference between internal irradiation and external irradiation, and the small sample size and short study time.\u003c/p\u003e\n\u003cp\u003eIn general, there are few literatures on the effect of gender and age on renal damage caused by radionuclide internal radiation therapy, and no unified opinion has been reached, and more studies are needed to confirm.\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eConflict of Interest:\u003c/h2\u003e\n\u003cp\u003eThe authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.\u003c/p\u003e\n\u003ch2\u003eInformed consent:\u003c/h2\u003e\n\u003cp\u003eThe animal experimentation has been approved by the ethics committee of Tianjin Medical University General Hospital.\u003c/p\u003e\n\u003ch2\u003eFunding:\u003c/h2\u003e\n\u003cp\u003eNo Funding.\u003c/p\u003e\n\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\n\u003cp\u003eLiang Yin and Yangyang Lin wrote the main manuscript text. Jianlan Yue and Zhinchun Lin prepared figures. All authors reviewed the manuscript.\u003c/p\u003e\n\u003ch2\u003eAcknowledgement\u003c/h2\u003e\n\u003cp\u003eWe are indebted to Weilong Li, Xiaolan Lv, and the technical staff of the Division of Nuclear Medicine of Tianjin Medical University General Hospital for their technical assistance and support.\u003c/p\u003e\n\u003ch2\u003eData Availability\u003c/h2\u003e\n\u003cp\u003eThe datasets used and/or analysed during the current study available from the corresponding author on reasonable request.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eFuma S, Kubota Y, Ihara S, et al. Radiocaesium contamination of wild boars in Fukushima and surrounding regions after the Fukushima nuclear accident [J]. J Environ Radioact, 2016, 164: 60-64.\u003c/li\u003e\n\u003cli\u003eStolniceanu CR, Nistor I, Bilha SC, et al. 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PLoS Pathog, 2020, 16(2): e1008297.\u003c/li\u003e\n\u003cli\u003eHe X, Xie Z, Dong Q, et al. Apoptosis in the kidneys of rats that experienced intrauterine growth restriction [J]. Nephrology (Carlton), 2015, 20(1): 34-39.\u003c/li\u003e\n\u003cli\u003eSchafer B, Quispe J, Choudhary V, et al. Mitochondrial outer membrane proteins assist Bid in Bax-mediated lipidic pore formation [J]. Mol Biol Cell, Mol Biol Cell, 2009, 20(8): 2276-2285.\u003c/li\u003e\n\u003cli\u003eKim H G, Jang S S, Lee J S, et al. Panax ginseng Meyer prevents radiation-induced liver injury via modulation of oxidative stress and apoptosis [J]. J Ginseng Res, 2017, 41(2): 159-168.\u003c/li\u003e\n\u003cli\u003eHudson D, Kovalchuk I, Koturbash I, et al. Induction and persistence of radiation-induced DNA damage is more pronounced in young animals than in old animals[J]. Aging (Albany NY), 2011, 3(6): 609-620.\u003c/li\u003e\n\u003cli\u003eUtsuyama M, Hirokawa K. Radiation-induced-thymic lymphoma occurs in young, but not in old rats [J]. Exp Mol Pathol, Exp Mol Pathol, 2003, 74(3): 319-325.\u003c/li\u003e\n\u003cli\u003eGrande T, Bueren J A. Involvement of the bone marrow stroma in the residual hematopoietic damage induced by irradiation of adult and young rats [J]. Exp Hematol, 1994, 22(13): 1283-1287.\u003c/li\u003e\n\u003cli\u003eNikkels P G, de Jong J P, Ploemacher R E. Radiation sensitivity of hemopoietic stroma: long-term partial recovery of hemopoietic stromal damage in rats treated during growth [J]. Radiat Res, 1987, 109(2): 330-341.\u003c/li\u003e\n\u003cli\u003eStojkovic R, Fucic A, Ivankovic D, et al. Age and sex differences in genome damage between prepubertal and adult rats after exposure to ionising radiation [J]. Arh Hig Rada Toksikol, 2016, 67(4): 297-303.\u003c/li\u003e\n\u003cli\u003eTachibana H, Morioka T, Daino K, et al. Early induction and increased risk of precursor B-cell neoplasms after exposure of infant or young-adult rats to ionizing radiation [J]. J Radiat Res, 2020, 61(5): 648-656.\u003c/li\u003e\n\u003cli\u003eFujimichi Y, Sasaki M, Yoshida K, et al. Effects of irradiation on cumulative mortality in rats: shifting toward a younger age of death [J]. J Radiat Res. 2023, 64(2): 412-419.\u003c/li\u003e\n\u003cli\u003eMilano MT, Marks LB, Olch AJ, et al. Comparison of Risks of Late Effects From Radiation Therapy in Children Versus Adults: Insights From the QUANTEC, HyTEC, and PENTEC Efforts [J]. Int J Radiat Oncol Biol Phys, 2024, 119(2): 387-400. \u003c/li\u003e\n\u003cli\u003eSaracyn M, Durma AD, Bober B, et al. Long-Term Complications of Radioligand Therapy with Lutetium-177 and Yttrium-90 in Patients with Neuroendocrine Neoplasms [J]. Nutrients, 2022, 15(1): 185.\u003c/li\u003e\n\u003cli\u003eVarma C, Schroeder MK, Price BR, et al. Long-Term, Sex-Specific Effects of GCRsim and Gamma Irradiation on the Brains, Hearts, and Kidneys of rats with Alzheimer\u0026apos;s Disease Mutations [J]. Int J Mol Sci, 2024, 25(16): 8948.\u003c/li\u003e\n\u003cli\u003eHolmes-Hampton GP, Soni DK, Kumar VP, et al. Time- and sex-dependent delayed effects of acute radiation exposure manifest via miRNA dysregulation [J]. iScience, 2024, 27(2): 108867. \u003c/li\u003e\n\u003cli\u003eKrukowski K, Grue K, Frias ES, et al. Female rats are protected from space radiation-induced maladaptive responses [J]. Brain Behav Immun. 2018, 74: 106-120.\u003c/li\u003e\n\u003cli\u003eApplegate K E, Ruhm W, Wojcik A, et al. Individual response of humans to ionising radiation: governing factors and importance for radiological protection [J]. Radiat Environ Biophys, 2020, 59(2): 185-209.\u003c/li\u003e\n\u003cli\u003eBodei L, Kidd M, Paganelli G, et al. Long-term tolerability of PRRT in 807 patients with neuroendocrine tumours: the value and limitations of clinical factors [J]. Eur J Nucl Med Mol Imaging, 2015, 42(1): 5-19. \u003c/li\u003e\n\u003cli\u003eNilica B, Svirydenka A, Fritz J, et al. Nephrotoxicity and hematotoxicity one year after four cycles of peptide receptor radionuclide therapy (PRRT) and its impact on future treatment planning - A retrospective analysis [J]. Rev Esp Med Nucl Imagen Mol (Engl Ed), 2022, 41(3): 138-145.\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Tables","content":"\u003cp\u003eTable 1: Mean optical density values of Caspase-3, Bcl-2 and Bax and Bcl-2/Bax ratio in kidney of rats of different age groups\u003cem\u003e(\u003c/em\u003e\u003cem\u003e\u0026nbsp;x̄ \u0026plusmn;s)\u003c/em\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"624\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 239px;\"\u003e\n \u003cp\u003eGrouping\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eCaspase-3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 98px;\"\u003e\n \u003cp\u003eBax\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 97px;\"\u003e\n \u003cp\u003eBcl-2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 94px;\"\u003e\n \u003cp\u003eBcl-2/Bax\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" style=\"width: 95px;\"\u003e\n \u003cp\u003eaged group\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 144px;\"\u003e\n \u003cp\u003eControl group\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003e21.18\u0026plusmn;2.27\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 98px;\"\u003e\n \u003cp\u003e16.25\u0026plusmn;2.34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 97px;\"\u003e\n \u003cp\u003e31.46\u0026plusmn;5.55\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 94px;\"\u003e\n \u003cp\u003e1.97\u0026plusmn;0.37\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 144px;\"\u003e\n \u003cp\u003eExperimental group\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003e27.82\u0026plusmn;5.78\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 98px;\"\u003e\n \u003cp\u003e20.78\u0026plusmn;3.66\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 97px;\"\u003e\n \u003cp\u003e26.24\u0026plusmn;6.18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 94px;\"\u003e\n \u003cp\u003e1.27\u0026plusmn;0.25\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" style=\"width: 95px;\"\u003e\n \u003cp\u003eYoung group\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 144px;\"\u003e\n \u003cp\u003eControl group\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003e21.96\u0026plusmn;4.97\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 98px;\"\u003e\n \u003cp\u003e17.03\u0026plusmn;4.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 97px;\"\u003e\n \u003cp\u003e32.56\u0026plusmn;7.52\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 94px;\"\u003e\n \u003cp\u003e2.05\u0026plusmn;0.81\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 144px;\"\u003e\n \u003cp\u003eExperimental group\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003e34.96\u0026plusmn;8.49\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 98px;\"\u003e\n \u003cp\u003e24.67\u0026plusmn;3.75\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 97px;\"\u003e\n \u003cp\u003e20.87\u0026plusmn;5.68\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 94px;\"\u003e\n \u003cp\u003e0.87\u0026plusmn;0.28\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eThe expression levels of Caspase-3, Bax, Bcl-2 and Bcl-2/Bax were significantly different between the control group and the experimental group (t =3.705, 3.615, -2.178, -5.373, \u003cem\u003erespectively\u003c/em\u003e; \u003cem\u003eP\u003c/em\u003e =0.001, 0.002, 0.040, 0.000, respectively).\u003c/p\u003e\n\u003cp\u003eThere were significant differences in Caspase-3, Bax, Bcl-2 and Bcl-2/Bax between the control group and the experimental group (t =4.579, 4.782, -4.297, -4.813, respectively; \u003cem\u003eall P\u003c/em\u003e =0.000).\u003c/p\u003e\n\u003cp\u003eThere were no significant differences in Caspase-3, Bax, Bcl-2 and Bcl-2/Bax ratio between the aged control group and the young control group (t =-0.492, -0.580, -0.404, -0.347, \u003cem\u003erespectively\u003c/em\u003e; \u003cem\u003eP\u003c/em\u003e =0.627, 0.568, 0.690, 0.732, respectively).\u003c/p\u003e\n\u003cp\u003eThere were significant differences in Caspase-3, Bax, Bcl-2 and Bcl-2/Bax ratio between the aged experimental group and the young experimental group (t =-2.406, -2.567, 2.217, 3.749, \u003cem\u003erespectively\u003c/em\u003e; \u003cem\u003eP\u003c/em\u003e =0.025, 0.018, 0.037, 0.001, respectively).\u003c/p\u003e\n\u003cp\u003eTable 2: Mean optical density values of Caspase-3, Bcl-2 and Bax and Bcl-2/Bax ratio in kidney of rats of different gender groups\u003cem\u003e(\u003c/em\u003e x̄ \u0026plusmn;s\u003cem\u003e)\u003c/em\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"624\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 249px;\"\u003e\n \u003cp\u003eGrouping\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eCaspase-3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003eBax\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 92px;\"\u003e\n \u003cp\u003eBcl-2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 92px;\"\u003e\n \u003cp\u003eBcl-2/Bax\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" style=\"width: 96px;\"\u003e\n \u003cp\u003eMale group\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 153px;\"\u003e\n \u003cp\u003eControl group\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003e21.66\u0026plusmn;3.98\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e16.71\u0026plusmn;3.62\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 92px;\"\u003e\n \u003cp\u003e32.44\u0026plusmn;6.33\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 92px;\"\u003e\n \u003cp\u003e2.05\u0026plusmn;0.69\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 153px;\"\u003e\n \u003cp\u003eExperimental group\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003e31.73\u0026plusmn;7.41\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e22.88\u0026plusmn;3.43\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 92px;\"\u003e\n \u003cp\u003e23.96\u0026plusmn;6.45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 92px;\"\u003e\n \u003cp\u003e1.07\u0026plusmn;0.32\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" style=\"width: 96px;\"\u003e\n \u003cp\u003eFemale group\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 153px;\"\u003e\n \u003cp\u003eControl group\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003e21.47\u0026plusmn;3.79\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e16.57\u0026plusmn;3.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 92px;\"\u003e\n \u003cp\u003e31.59\u0026plusmn;6.89\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 92px;\"\u003e\n \u003cp\u003e1.97\u0026plusmn;0.56\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 153px;\"\u003e\n \u003cp\u003eExperimental group\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003e31.06\u0026plusmn;8.83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e22.57\u0026plusmn;4.89\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 92px;\"\u003e\n \u003cp\u003e23.15\u0026plusmn;6.65\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 92px;\"\u003e\n \u003cp\u003e1.07\u0026plusmn;0.35\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eThere were significant differences in Caspase-3, Bax, Bcl-2 and Bcl-2/Bax ratio between the control group and the experimental group in male rats (t =4.143, 4.294, -3.252, -4.457, \u003cem\u003erespectively\u003c/em\u003e; \u003cem\u003eP\u003c/em\u003e =0.000, 0.000, 0.004, 0.000, respectively).\u003c/p\u003e\n\u003cp\u003eThere were significant differences in Caspase-3, Bax, Bcl-2 and Bcl-2/Bax between the control group and the experimental group in female rats (t =3.454, 3.607, -3.047, -4.712; \u003cem\u003eP\u003c/em\u003e =0.002, 0.002, 0.006, 0.000, \u003cem\u003erespectively\u003c/em\u003e).\u003c/p\u003e\n\u003cp\u003eThere were no significant differences in Caspase-3, Bax, Bcl-2 and Bcl-2/Bax between male control group and female control group (t =0.120, 0.098, 0.319, 0.325, \u003cem\u003erespectively\u003c/em\u003e; \u003cem\u003eP\u003c/em\u003e =0.906, 0.923, 0.753, 0.749, respectively).\u003c/p\u003e\n\u003cp\u003eThere were no significant differences in Caspase-3, Bax, Bcl-2 and Bcl-2/Bax between male and female rats (t =0.202, 0.181, 0.302, -0.013, \u003cem\u003erespectively\u003c/em\u003e; \u003cem\u003eP\u003c/em\u003e =0.842, 0.858, 0.765, 0.990, respectively).\u003c/p\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":"radioactive iodine, age, gender, renal cell, apoptosis, rats","lastPublishedDoi":"10.21203/rs.3.rs-8555726/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8555726/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eObjective:\u003c/strong\u003e To explore the effects of age and gender on renal cell apoptosis induced by high-dose radioactive iodine (\u003csup\u003e131\u003c/sup\u003eI) in rats.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods:\u003c/strong\u003e Forty-eight rats were divided into 4 groups (aged male group, aged female group, young male group, and young female group) according to age and sex, with 12 rats in each group, each group was randomly divided into control group and experimental group, with a total of 8 groups and 6 rats in each group. All rats in the experimental groups were intraperitoneally injected with 1.85 MBq/2g \u003csup\u003e131\u003c/sup\u003eI solution, and rats in control group were intraperitoneally injected with the same volume of 0.9% NaCl solution. All the rats were sacrificed after 4 weeks, the kidney sections were removed for immunohistochemical analysis of apoptosis-related proteins such as Bax, Bcl-2 and Caspase-3.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults: \u003c/strong\u003e4 weeks later, the expression levels of Caspase-3 and Bax proteins in all the experimental groups were significantly increased, while the expression level of Bcl-2 protein and Bcl-2/Bax ratio were significantly decreased. Compared with the aged rats, the expression level of apoptosis in young rats was increased more significantly, and the difference was statistically significant. There was no significant difference in the apoptosis level between male and female rats.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion:\u003c/strong\u003e A single high-dose of \u003csup\u003e131\u003c/sup\u003eI can lead to a significant increase in the level of renal cell apoptosis in rats, and the level of apoptosis is more obvious in young rats than aged rats, but the gender of the rats is not related to the level of apoptosis.\u003c/p\u003e","manuscriptTitle":"Effect of age and gender on radioactive iodine induced apoptosis in rats kidney cells","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-02-17 09:12:06","doi":"10.21203/rs.3.rs-8555726/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":"45e9a843-34b4-4b06-a81d-e3096ed99111","owner":[],"postedDate":"February 17th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":62754303,"name":"Biological sciences/Biochemistry"},{"id":62754304,"name":"Health sciences/Nephrology"},{"id":62754305,"name":"Biological sciences/Physiology"}],"tags":[],"updatedAt":"2026-03-05T03:54:52+00:00","versionOfRecord":[],"versionCreatedAt":"2026-02-17 09:12:06","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8555726","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8555726","identity":"rs-8555726","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}