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目的:探讨患糖尿病12周大鼠睾丸生精细胞周期及其凋亡和血清与睾丸抗氧化水平的变化。方法:W istar大鼠随机分为正常对照组10只,糖尿病组20只。腹腔注射链佐脲菌素(STZ)建立糖尿病大鼠模型,12周末记录其存活率、体重和睾丸重量;采用流式细胞术检测生精细胞周期各时相细胞百分率和细胞凋亡率,应用硫代巴比妥酸法(TBAR s)、硝酸还原酶法、黄嘌呤氧化酶法、二硫代二硝基苯甲酸法(DTNB)和分光光度法分别检测血清及睾丸丙二醛(MDA)和一氧化氮(NO)含量,超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)和NO合酶(NOS)活性。结果:大鼠患糖尿病12周后,其存活率、体重和睾丸重量显著低于正常对照组(P<0.05);G0/G1期生精细胞显著增加(P<0.05),S期和G2/M期细胞减少,即发生了G0/G1期细胞阻滞;生精细胞凋亡率明显增加(P<0.05)。与正常对照组相比,糖尿病大鼠血清和睾丸MDA含量增加,其中后者增加明显(P<0.01);血清及睾丸SOD活性降低;血清GSH-Px活性显著低于对照组(P<0.05),而睾丸GSH-Px活性显著增高(P<0.01);血清和睾丸NO含量增高,尤其前者显著升高(P<0.01);血清NOS活性显著降低(P<0.05)。结论:睾丸组织及血清MDA和NO含量增加,以及抗氧化酶活性降低,可能与糖尿病大鼠生精细胞G0/G1期阻滞和凋亡增多所致生精障碍有关。
Objective: To investigate the changes of spermatogenic cell cycle, apoptosis and serum and testicular antioxidant levels in the testes of diabetic rats for 12 weeks. Methods: Wistar rats were randomly divided into normal control group (n = 10) and diabetic group (n = 20). The model of diabetic rats was established by intraperitoneal injection of streptozotocin (STZ), and the survival rate, body weight and testicular weight were recorded at the end of 12th week. The percentage of cells in each phase of germ cell cycle and apoptosis rate were detected by flow cytometry Thiobarbituric acid (TBARs), nitrate reductase, xanthine oxidase, DTNB and spectrophotometry were used to detect serum and testis malondialdehyde (MDA) And nitric oxide (NO) content, superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and NO synthase (NOS) activity. Results: After 12 weeks, the survival rate, body weight and testicular weight of diabetic rats were significantly lower than those of normal control group (P <0.05). The number of spermatogenic cells in G0 / G1 phase increased significantly (P <0.05) The number of senescent cells decreased, that is, G0 / G1 phase cell arrest occurred. The apoptosis rate of spermatogenic cells was significantly increased (P <0.05). Compared with the normal control group, the content of MDA in serum and testis of diabetic rats increased, the latter increased significantly (P <0.01), the activity of SOD in serum and testis decreased, the activity of GSH-Px in serum decreased significantly (P <0.05) , While the activity of GSH-Px in testis increased significantly (P <0.01). The content of NO in serum and testis increased, especially the former increased significantly (P <0.01) and the activity of NOS decreased significantly (P <0.05). CONCLUSION: Increased MDA and NO contents in testis, serum and antioxidase activity may be related to dysgenesis of spermatogenesis induced by increased G0 / G1 arrest and apoptosis in germ cells of diabetic rats.