基于TGF-β1/Smads信号通路观察强精胶囊对精索静脉曲张大鼠生育力的影响

Effect of Qiangjing Capsule on Fertility of Varicocele Rats Based on TGF-β1/Smads Signaling Pathway

目的:观察强精胶囊对精索静脉曲张(varicocele,VC)大鼠附睾组织中TGF-β1/Smads信号通路中的关键信号及大鼠生育力的影响,探讨强精胶囊治疗VC性不育的可能作用机制。方法:从68只青春期雄性SD大鼠中随机抽取8只为空白对照组,剩余60只利用缩窄左肾静脉成功建立VC模型后,随机分为模型对照组、强精胶囊组(高、中、低剂量)、五子衍宗+少腹逐瘀组。分别给予生理盐水、强精胶囊混悬液(高、中、低剂量)、五子衍宗胶囊+少腹逐瘀胶囊混悬液灌胃,每天一次,连续4周。利用Western blot和RT-PCR检测大鼠附睾组织中的TGF-β1蛋白水平与mRNA水平的表达以及Smad2、Smad7的蛋白水平表达,并观察大鼠精子质量的变化以及雌性大鼠与其合笼交配后的受孕情况。结果:相较于空白对照组,模型对照组精子密度与活力显著降低(P<0.01),TGF-β1的蛋白、mRNA表达均显著升高(P<0.01),Smad2的蛋白表达显著升高(P<0.01),Smad7的蛋白表达显著减少(P<0.01)。相较于模型对照组,强精胶囊(高、中、低剂量)及五子衍宗+少腹逐瘀组的精子密度与活力显著提高(P<0.01),TGF-β1、Smad2的蛋白表达均减少(P<0.05),Smad7的蛋白表达显著升高(P<0.01),TGF-β1的mRNA表达减弱(P<0.05)。与动情期雌性大鼠合笼后,发现高剂量强精胶囊组受孕率最高。结论:强精胶囊可以通过TGF-β1/Smads信号通路,下调VC大鼠附睾组织中TGF-β1的蛋白、mRNA表达以及Smad2的蛋白表达,上调Smad7的蛋白表达,提高VC大鼠精子质量以及受孕率,从而改善VC大鼠的生育力。

Objective:The objective of this study was to investigate the effects of Qiangjing capsules on the key signals in the TGF-β1/Smads signaling pathway in the epididymis of varicocele(VC)rats,as well as the fertility of rats.The possible mechanism of action of Qiangjing capsules in the treatment of VC infertility was also investigated.Methods:A total of 68 adolescent male SD rats were used in this study.Eight rats were randomly selected as the blank control group,while the remaining 60 rats were randomly divided into the model control group,Qiangjing capsule group(high,medium,and low doses),and Wuzi Yanzong+Shao Abdominal Expulsion group after the VC model was successfully established by narrowing the left renal vein.Each group was given saline,strong essence capsule suspension(high,medium,and low dose),and Wu Zi Yan Zong capsule+Shao Abdominal Expulsion capsule suspension by gavage once a day for 4 weeks.Western blot and RT-PCR were used to detect the protein level and mRNA level expression of TGF-β1 and the protein level expression of Smad2 and Smad7 in the epididymal tissues of rats.The changes in sperm quality of rats and the conception of female rats after mating with them in a combined cage were also observed.Results:The model control group showed significantly lower sperm density and viability(P<0.01),significantly higher protein and mRNA expression of TGF-β1(P<0.01),significantly higher protein expression of Smad2(P<0.01),and significantly lower protein expression of Smad7(P<0.01)compared with the blank control group.Compared with the model control group,sperm density and viability were significantly increased in the Qiangjing Capsules(high,medium,and low doses)and the Wuzi Yan Zong+Shao Abdominal Expulsion group(P<0.01).Protein expression of TGF-β1 and Smad2 were reduced(P<0.05),protein expression of Smad7 was significantly increased(P<0.01),and mRNA expression of TGF-β1 was attenuated(P<0.05).The highest conception rate was found in the high-dose strong semen capsule group after cage combination with female rats in the motility phase.Conclusion:Qiangjing capsules can improve the fertility of VC rats by down-regulating the protein and mRNA expression of TGF-β1 and Smad2 and up-regulating the protein expression of Smad7 in the epididymis of VC rats through the TGF-β1/Smads signaling pathway.This finding suggests that Qiangjing capsules can improve the sperm quality and conception rate of VC rats.