龟鹿二仙胶对雷公藤多苷诱导弱精子症大鼠精子线粒体结构和功能及SLC22A14表达的影响

Effects of Guilu-Erxian-Glue on the structure and function of sperm mitochondria and the expression of SLC22A14 in asthenospermia rat model induced by Tripterygium wilfordii polyglycoside

目的:研究龟鹿二仙胶对雷公藤多苷(GTW)诱导弱精子症(AZS)大鼠精子线粒体结构和功能及溶质载体家族22成员14(SLC22A114)表达的影响。方法:60只SD大鼠随机分为正常组,模型组,龟鹿二仙胶低、中、高剂量组、五子衍宗丸组,各10只;除正常组外,各组均给予GTW40mg·kg^(-1)·d^(-1)连续灌胃4周造模AZS大鼠模型;造模结束后正常组、模型组给予0.9%氯化钠溶液灌胃4周,龟鹿二仙胶低、中、高剂量组分别给予龟鹿二仙胶混悬液0.25、0.50、1.00g·kg^(-1)·d^(-1),五子衍宗丸组给予1.00g·kg^(-1)·d^(-1)五子衍宗丸混悬液,灌胃4周;采用HE染色观察睾丸组织形态;计算机辅助精子分析检测大鼠精液质量;流式细胞术分析检测精子线粒体膜电位;透射电镜观察精子线粒体结构;实时荧光定量聚合酶链式反应和蛋白免疫印迹法检测精子线粒体SLC22A114蛋白及mRNA表达。结果:龟鹿二仙胶可减轻AZS大鼠模型睾丸组织病理损伤,改善精液质量,且龟鹿二仙胶高剂量组可显著改善精子浓度、精子活力、活动精子数(P<0.01);龟鹿二仙胶可提高AZS大鼠精子线粒体膜电位(P<0.05,P<0.01),并可改善其结构;龟鹿二仙胶高剂量组可上调AZS大鼠精子线粒体SLC22A14蛋白及mRNA表达(P<0.05,P<0.01)。结论:龟鹿二仙胶可通过上调GTW诱导AZS大鼠模型精子线粒体SLC22A14蛋白及mRNA表达,进而改善精子线粒体结构和功能,提高精子活力。

Objective:To study the effects of Guilu-Erxian-Glue on the structure and function of sperm mitochondria and the expression of solute carrier family 22 member 14(SLC22A114)in asthenospermia(AZS)rat model induced by Tripterygium wilfordi polyglycosid(GTW).Methods:Sixty SD rats were randomly divided into normal group,model group,Guilu-Erxian-Glue low,medium and high dose group and Wuzi-Yanzong-Pill group,with 10 rats in each group;In addition to the normal group,each group was given Tripterygium wilfordi polyglycoside 40 mg·kg^(-1)·d^(-1)for 4 weeks to induce the AZS rat model;After modeling,the normal and model group were given 0.9%sodium chloride solution by gavage for 4 weeks,and the Guilu-Erxian-Glue low,medium,high group were given 0.25,0.50,1.00 g·kg^(-1)·d^(-1)Guilu-Erxian-Glue,Wuzi-Yanzong-Pill group were given 1.00 g·kg^(-1)·d^(-1)Wuzi-Yanzong-il,suspension by gavage for 4 weeks;The morphology of testis was observed by HE staining;The sperm quality was detected by computer-aided sperm analysis;Sperm mitochondrial membrane potential was detected by flow cytometry;The morphology of sperm mitochondria were observed by transmission electron microscope;The protein and mRNA expressions of SLC22A14 of sperm mitochondria were detected by real-time PCR and western blot.Results:Guilu-Erxian-Glue could mitigate the pathological damage of testicular tissue and improve the sperm quality of epididymis in the AzS rat model,Guilu-Erxian-Glue high dose group could improve the sperm concentration,sperm motility,and mobile sperm counts(P<0.01);Guilu-Erxian-Glue could increase the sperm mitochondrial membrane potential(P<0.05,P<0.01)and improve its structure;Guilu-Erxian-Glue high dose group could up-regulate the protein and mRNA expressions of SLC22A14 of sperm mitochondria(P<0.05,P<0.01).Conclusion:Guilu-Erxian-Glue could up-regulate the expression of protein and mRNA of sperm mitochondria SLC22A14 in the AZS rat model induced by Tripterygium wilfordi polyglycoside,so as to improve the structure and function of sperm mitochondria,and improve the sperm motility.