白藜芦醇对2型糖尿病大鼠睾丸PCNA、SOD表达及生精功能影响

Effects of Resveratrol on the Expression of PCNA and SOD in Testis of Type 2 Diabetic Rats and Its Effect on Spermatogenesis

目的 :研究白藜芦醇对2型糖尿病大鼠睾丸PCNA、SOD的表达及对生精功能的影响。方法 :首先培养2型糖尿病大鼠模型即实验组,先喂雄性SD大鼠高热量饲料4周来进行诱导胰岛素抵抗,然后给予小剂量STZ[链脲佐菌素(streptozocin)]连续2次进行腹腔注射,继续高热量饲料喂养诱导小鼠成2型糖尿病模型。将实验组随机分为6组:A组(n=10)、B组(n=10)、C组(n=10)及3组对照组(n=10)用于后面3项检测。对照组均不做处理。A组、B组和C组:将中药单体白藜芦醇(购于上海纳贝生物技术有限责任公司)分别用无菌蒸馏水稀释后进行灌胃。检测睾丸中SOD活性:A组:采用断头处死大鼠,取一侧睾丸组织来称重、摇匀浆液后按试剂盒说明采用黄嘌呤氧化酶法进行操作测定。检测PCNA表达的情况:B组:大鼠麻醉通过腹腔注射,固定选用4%多聚甲醛灌注,取双侧睾丸,固定一夜后进行切片处理,单片厚5μm,PCNA的表达情况通过SP免疫组化法来检测。C组:对睾丸质量、睾酮含量、黄体生成素及ECFR含量进行检测。结果 :睾丸组织中SOD活性的检测及对睾丸组织中PCNA表达的检测,说明白藜芦醇对2型糖尿病大鼠SOD活性提高和PCNA表达有很好的效果,两组检测比较差异具有统计学意义(P<0.05)。白藜芦醇治疗后睾丸质量、睾酮含量、黄体生成素及ECFR含量较治疗前明显提高,两者比较差异显著,具有统计学意义(P<0.05)。对照组小鼠精子密度、精子活率显著低于正常组,差异具有统计学意义(P<0.05)。对照组小鼠精子密度、精子活率显著低于C组,差异具有统计学意义(P<0.05)。白藜芦醇可显著增加小鼠的精子密度及精子存活率。白藜芦醇均可显著恢复小鼠各级生精细胞的百分比,差异具有统计学意义(P<0.05)。结论 :通过对小鼠治疗前后的比较,可以看出白藜芦醇对2型糖尿病大鼠SOD活性提高和PCNA表达有很好的效果。小鼠在经过白藜芦醇治疗后,睾丸质量、睾酮含量、黄体生成素及ECFR均有明显提高。且白藜芦醇显著增加小鼠的精子密度及精子存活率,还可以显著恢复小鼠各级生精细胞的百分比,对提高生精功能有重要影响,治疗前、后比较差异显著,有助于延缓衰老,改善生理功能,对生精功能的改善有较好的作用。

Objective ：To study the effect of resveratrol on the expression of PCNA and SOD in testis of type 2 diabetic rats and its effect on spermatogenesis.Methods ：The male rats were fed with highcalorie diet for 4 weeks to induce insulin resistance,and then low-dose STZ（streptozocin）（30 mg/kg）was administered continuously.2 times intraperitoneal injection,continue to high calorie feed feeding induced mode.The experimental group was randomly divided into 6 groups ：group A（n=10）,group B（n=10）,group C（n=10）and group 3 control group（n=10）for the following three tests.The control group was not treated.Group A,group B and group C ：Traditional Chinese medicine resveratrol（purchased from Shanghai Beibei biotechnology Co.,Ltd.）was diluted with sterile distilled water and then fed.Detection of SOD activity in testis ：A group ：the rats were sacrificed and the rats were sacrificed.The rats were sacrificed and the testis tissues were taken quickly.After weighing,homogenization was performed by xanthine oxidase method according to the instructions of the kit.PCNA expression ：B group ：rats were intraperitoneally anesthetized,4% paraformaldehyde perfusion fixed,take bilateral testes,fixed overnight after the slice,slice thickness 5 μm,according to the kit to SP immunohistochemical detection of PCNA expression.Group C ：testicular mass,testosterone content,luteinizing hormone and ECFR content were detected.Results ：The detection of SOD activity in the testis tissue and the detection of PCNA expression in the testis tissue showed that the effect of resveratrol on SOD activity and PCNA expression in type 2 diabetic rats was very good.The difference between the two groups was statistically significant P〈0.05）.The content of testis,testosterone,luteinizing hormone.The sperm density and sperm viability were significantly lower in the control group than in the C group,the difference was statistically significant（P〈0.05）.Resveratrol can significantly increase sperm density and sperm viability in mice.Resveratrol could significantly restore the percentage of spermatogenic cells in mice at all levels.Conclusion ：Through the comparison of the mice before and after treatment,we can see that observing the effect of resveratrol on SOD activity and PCNA expression in type 2 diabetic rats has a good effect.The testis mass,testosterone content,luteinizing hormone and ECFR were significantly increased after treatment with resveratrol.And resveratrol significantly increased the sperm density and sperm survival rate of mice,but also can significantly restore the percentage of spermatogenic cells in mice at all levels,to improve spermatogenic function has an important impact,before and after treatment has significantly different.In the anti-aging,improving the physiological function of the improvement of spermatogenic function has a better effect.