| 摘要: |
| 目的 探讨自噬在大鼠睾丸扭转所致缺血再灌注损伤（I/RI）中的作用机制。方法 将40只健康雄性SD大鼠随机分为假手术组、I/RI组、I/RI＋雷帕霉素（RAPA）组和I/RI＋3-甲基腺嘌呤（3-MA）组，每组10只。后3组大鼠建立左侧睾丸扭转（720°，1 h）复位模型，I/RI＋RAPA组和I/RI＋3-MA组大鼠造模前分别给予自噬激动剂RAPA和自噬抑制剂3-MA干预。于睾丸复位12 h后取各组大鼠左侧睾丸和附睾，收集精子进行精子质量分析，采用H-E染色观察睾丸组织病理变化，采用试剂盒检测睾丸组织中超氧化物歧化酶（SOD）活性、丙二醛（MDA）含量、总抗氧化能力（T-AOC），采用TUNEL染色检测睾丸组织细胞凋亡水平，通过蛋白质印迹法检测睾丸组织中自噬相关蛋白（Beclin1、p62）和凋亡相关蛋白（Bcl2、Bax、cleaved caspase 3）表达水平。结果 与假手术组比较，I/RI组大鼠精子质量下降，睾丸曲细精管排列松散，生殖细胞减少、脱落，睾丸组织SOD活性、T-AOC降低（均P<0.05），MDA含量升高（P<0.05），p62和Bcl2蛋白表达水平降低（均P<0.05），Beclin1、Bax和cleaved caspase 3蛋白表达水平升高（均P<0.05），凋亡生殖细胞明显增多。与I/RI组比较，I/RI＋RAPA组大鼠精子质量提高，睾丸曲细精管排列正常，睾丸组织SOD活性、T-AOC升高（均P<0.05），MDA含量降低（P<0.05），Beclin1和Bcl2蛋白表达水平升高（均P<0.05），p62、Bax和cleaved caspase 3蛋白表达水平降低（均P<0.05），凋亡生殖细胞减少；I/RI＋3-MA组大鼠精子质量下降，睾丸曲细精管排列松散，睾丸组织SOD活性、T-AOC降低（均P<0.05），MDA含量升高（P<0.05），Beclin1和Bcl2蛋白表达水平降低（均P<0.05），p62、Bax和cleaved caspase 3蛋白表达水平升高（均P<0.05），凋亡生殖细胞增多。结论 激活自噬可改善I/RI大鼠精子质量和睾丸组织病理损伤，缓解氧化应激，抑制生殖细胞凋亡。 |
| 关键词: 睾丸扭转 缺血再灌注损伤 自噬 氧化性应激 生殖细胞 细胞凋亡 |
| DOI：10.16781/j.CN31-2187/R.20230051 |
| 投稿时间：2023-02-15修订日期：2023-07-10 |
| 基金项目:武汉市卫生健康委员会面上项目重点项目（WX20A16）. |
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| Role of autophagy in ischemia/reperfusion injury caused by testicular torsion in rats |
| HU Zhi1,XU Lü2,SUN Wei2,FU Qiao2,ZHANG Wei2,CHEN Yiyan2,CHU Hao2* |
(1. Department of Urology, Guanggu Branch of Tongren Hospital of Wuhan University (Wuhan Third Hospital), Wuhan 430014, Hubei, China;
2. Department of Urology, Tongren Hospital of Wuhan University (Wuhan Third Hospital), Wuhan 430000, Hubei, China
*Corresponding author) |
| Abstract: |
| Objective To investigate the role of autophagy in ischemia/reperfusion injury (I/RI) caused by testicular torsion in rats. Methods Forty healthy male SD rats were randomly assigned (1:1:1) to sham group, model (I/RI) group, I/RI+rapamycin (RAPA) group, or I/RI+3-methyladenine (3-MA) group. Left testicular torsion (720°, 1 h) reduction rat models were established in the latter 3 groups. The rats in the I/RI+RAPA group and I/RI+3-MA group were intervened with autophagy agonist RAPA and autophagy inhibitor 3-MA before modeling, respectively. At 12 h after testicular reduction, the left testis and epididymis of rats in each group were collected for sperm quality analysis. The histopathological changes of testicular tissue were observed by hematoxylin-eosin (H-E) staining. The levels of superoxide dismutase (SOD), malondialdehyde (MDA), and total antioxidant capacity (T-AOC) in testicular tissue were measured by kits. The level of apoptosis in testicular tissue was detected by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay (TUNEL). The expression levels of autophagy-related proteins (Beclin1 and p62) and apoptosis-related proteins (Bcl2, Bax, and cleaved caspase 3) in testicular tissue were detected by Western blotting. Results Compared with the rats in the sham group, the rats in the I/RI group had poorer sperm quality, loosened testicular varicocele arrangement, fewer and shed germ cells, significantly lower SOD activity, T-AOC level, and p62 and Bcl2 expression levels (all P<0.05), significantly higher MDA content and expression levels of Beclin1, Bax, and cleaved caspase 3 (all P<0.05), and more apoptotic germ cells. Compared with the rats in the I/RI group, the rats in the I/RI+RAPA group had better sperm quality, normalized testicular varicocele arrangement, significantly higher SOD activity, T-AOC level, Beclin1 and Bcl2 expression levels (all P<0.05), significantly lower MDA content, p62, Bax and cleaved caspase 3 expression levels (all P<0.05), and fewer apoptotic germ cells; the rats in the I/RI+3-MA group had poorer sperm quality, loosened testicular varicocele arrangement, significantly lower SOD activity, T-AOC level, Beclin1 and Bcl2 expression levels (all P<0.05), significantly higher MDA content, p62, Bax and cleaved caspase 3 expression levels (all P<0.05), and more apoptotic germ cells. Conclusion Activation of autophagy can improve the sperm quality and testicular histopathological damage in I/RI rats, alleviate oxidative stress, and inhibit germ cell apoptosis. |
| Key words: testicular torsion ischemia/reperfusion injury autophagy oxidative stress germ cells apoptosis |
