加味补中益气汤对少弱精子症小鼠精液质量及线粒体功能的影响

摘要

目的:本实验通过观察加味补中益气汤对雷公藤多苷诱导少弱精子症小鼠的精液质量、线粒体膜电位和线粒体超微结构的影响,探讨加味补中益气汤治疗少弱精子症的作用机制.方法:以雷公藤多苷制备少弱精子症小鼠模型,随机分为正常组、模型组、对照组、加味补中益气汤低剂量组、加味补中益气汤高剂量组,每组12只,应用伟力精子测试仪检测小鼠精液质量,应用JC-1荧光染色流式细胞技术测定小鼠精子线粒体膜电位正常的精子百分率(JC-1+％),应用日立HT7700型透射电镜观察附睾精子超微结构.结果:对照组、中药组的A级、A+B级精子的百分比与模型组相比有统计学意义(P＜0.05);对照组与中药低剂量组相比有统计学意义(P＜0.05);中药高剂量组与对照组相比无统计学意义(P＞0.05).对照组、中药组的精子浓度、总数与模型组相比具有统计学意义(P＜0.05);对照组、中药低剂量组、中药高剂量组相互之间无统计学意义(P＞0.05).模型组小鼠精予JC-1+％较正常组显著下降(P＜0.05);中药高剂量组与中药低剂量组和对照组相比有显著性差异(P＜0.05);中药低剂量组与对照组相比有显著性差异(P＜0.05).正常组小鼠精子的尾部中段横切面可见完整的线粒体鞘结构.模型组整个精子线粒体肿胀明显、纤维明显疏松、轴丝断裂.药物干预后各组精子线粒体鞘结构较完整,肿胀较前显著减轻,纤维、轴丝结构较整齐、完整.结论:加味补中益气汤可以显著改善睾丸精子的发生及成熟,其可能的机制是通过调节小鼠精子线粒体功能(改善线粒体膜电位及超微结构)从而改善睾丸精子发生达到提高精子质量的目的.%Objectives:To discuss the mechanism of Jiawei Buzhong Yiqi Recipe in treating Oligoasthenospermia mice by bbserving the impact of Jiawei Buzhong Yiqi Recipe on such mice's semen quality,mitochondria membrane potential and ultrastructures of mitochondria.Methods:Oligoasthenospermia mice induced using tripterygium glycosides were randomly divided into normal group,model group,control group,low Jiawei Buzhong Yiqi Recipe dose group and high Jiawei Buzhong Yiqi Recipe dose group,each of 12 mice.WLJU-9000 Sperm tester was adopted to analyze sperm quality,JC-1 fluorescent staining and flow cytometry was used to detect the percentage of normal mitochondria membrane potential's sperm (JC-1 + ％),and Hitachi HT770 Transmission Electron Microscopy was used to observe the ultrastructure of epididymal sperm.Results:The percentage of class A,A + B sperm in the control group and Chinese medicine group had statistically significant difference compared with the model group (P ＜ 0.05);and the control group had statistically significant difference compared with the low dose group (P ＜ 0.05),but not with the high dose group (P ＞ 0.05).The total number of sperm and sperm concentration in the control group and the Chinese medicine group had statistically significant difference compared with model group (P ＜ 0.05);but there was no statistically significant difference between the control group,low dose group and high dose group.The JC-1 + ％ in model group decreased more significantly compared that in the normal group (P ＜ 0.05);high dose group and low dose group had significant difference compared with control group (P ＜ 0.05);low dose group had significant difference compared with control group (P ＜ 0.05).Complete mitochondrial sheath structure was found in the cross section of sperm tail in normal group.In the model group,the sperm's mitochondria were significant swollen,with significant loose fiber and ruptured axial filament.After drug intervention,each group sperm's mitochondrial sheath structure was complete,the swollen was significantly reduced,and axial filament became complete and neat.Conclusion:Jiawei Buzhong Yiqi Recipe can significantly improve the sperm's generation and mature,possibly through adjusting the sperm mitochondrial function in mice (improving the mitochondria membrane potential and ultrastructures of mitochondria) to improve the quality of sperm.