探讨活血养阴法调控Bcl-2/Bax通路改善精索静脉曲张大鼠生精功能的作用机制  

Exploring the mechanism of regulating Bcl-2/Bax pathway by the therapy of promoting blood circulation and nourishing yin to improve spermatogenesis in rats with varicocele

作　　者：梁仪春 王文 梁树麟 付瑞杰 覃湛[2] 耿立果[2] 何超拔 袁少英[2] LIANG Yichun;WANG Wen;LIANG Shulin;FU Ruijie;QIN Zhan;GENG Liguo;HE Chaoba;YUAN Shaoying(The Second Clinical Medical College,Guangzhou University of Chinese Medicine,Guangzhou 510006,China;Zhuhai Hospital,Guangdong Provincial Hospital of Chinese Medicine,Zhuhai 510915,China)

机构地区：[1]广州中医药大学第二临床医学院,广州510006 [2]广东省中医院珠海医院

出　　处：《北京中医药大学学报》2024年第11期1481-1489,共9页Journal of Beijing University of Traditional Chinese Medicine

基　　金：广东省中医药局中医药科研项目面上项目(No.202205111913416600);珠海市科技局项目(No.2220004000307,No.2320004000300)。

摘　　要：目的基于B淋巴细胞瘤-2(Bcl-2)/Bcl-2相关X蛋白(Bax)通路探讨活血养阴法改善精索静脉曲张大鼠生精功能的作用机制。方法选取8周龄SD雄性大鼠30只,按随机数字表法分为假手术组、模型组、左卡尼汀组(1.0 g/kg)、活血养阴法低剂量组(5.2 g/kg)、活血养阴法中剂量组(10.4 g/kg)及活血养阴法高剂量组(20.8 g/kg),每组5只。除假手术组外,其余大鼠均通过经典的Turner法复制精索静脉曲张模型。术后4周,假手术组及模型组大鼠灌胃生理盐水,各给药组灌胃相应药物,每日1次,连续4周。灌胃结束后,使用计算机辅助精子分析仪检测精子质量,HE染色法观察睾丸组织病理学形态并通过Johnsen评分评价睾丸生精功能,TUNEL法检测睾丸细胞凋亡情况,实时荧光PCR法检测睾丸组织Bcl-2及Bax mRNA表达,免疫组织化学法检测细胞色素C蛋白表达。结果与假手术组比较,模型组大鼠的精子总数、精子活动率及(a+b)级精子率均降低(P<0.05);睾丸组织中生精小管排列紊乱,有明显空泡样改变,Johnsen评分下降(P<0.05);睾丸组织细胞凋亡率增加(P<0.05);睾丸组织Bax mRNA表达上升(P<0.05);细胞色素C蛋白阳性表达增加(P<0.05)。与模型组比较,活血养阴法中剂量组大鼠的精子总数,低、中剂量组精子活动率,以及低剂量组(a+b)级精子率均上升(P<0.05);大鼠睾丸病理学结构有不同程度的改善,活血养阴法中剂量组Johnsen评分增加(P<0.05);左卡尼汀组与活血养阴法各剂量组睾丸细胞凋亡率均下降(P<0.05);活血养阴法低、高剂量组Bcl-2 mRNA表达上升(P<0.05),左卡尼汀组与活血养阴法中剂量组Bax mRNA表达下降(P<0.05);活血养阴法中剂量组细胞色素C蛋白阳性表达下降(P<0.05)。结论活血养阴法可提高精索静脉曲张大鼠的精子质量,修复损伤的睾丸组织结构,改善生精功能,其作用机制可能与激活Bcl-2/Bax凋亡通路、抑制细胞凋亡有关。Objective To explore the mechanism of the therapy of promoting blood circulation and nourishing yin(PBCNY)in improving spermatogenesis in rats with varicocele based on Bcl-2/Bax pathway.Methods Thirty 8-week-old male SD rats were randomly divided into six groups(five rats per group):sham,model,levocarnitine(1.0 g/kg),and PBCNY low-dose(5.2 g/kg),PBCNY mediumdose(10.4 g/kg),and PBCNY high-dose(20.8 g/kg)groups.Apart from the sham group,all the groups were built a varicocele model by the classical Turner method.After 4 weeks,the sham group and model group were gavaged with normal saline,and the remaining groups were given with levocarnitine or different doses of PBCNY decoction once a day for 4 weeks.At the end of gavage,the sperm quality of rats in each group was detected by a computer-assisted sperm analyzer,and HE staining was used to observe the testicular tissue morphology of rats in each group and performed with Johnsen score.TUNEL method was used to detect apoptosis,RT-qPCR was used to detect Bcl-2 and Bax mRNA expression,and immunohistochemical method was used to detect cytochrome C protein expression.Results Compared to the sham group,sperm total count,motility rate and percentage of sperm with grade(a+b)motility were lower in the model group of rats(P<0.05).HE staining showed disturbed arrangement of seminiferous tubules in the testicular tissue with significant changes and decreased Johnsen score(P<0.05).The result of TUNEL assay showed that the apoptosis rate was increased in the model group(P<0.05).Bax mRNA expression was increased in testicular tissue(P<0.05).The result of immunohistochemistry showed that the positive expression of cytochrome C was increased in the model group(P<0.05).Compared with the model group,the rat sperm total count in the PBCNY medium-dose group,motility rate in the PBCNY low-and medium-dose groups,and percentage of sperm with grade(a+b)motility in the PBCNY low-dose group increased(P<0.05);the pathological structure of rat testis had different degrees of improvement,and Johns

关 键 词：活血养阴法 精索静脉曲张 生精功能 细胞凋亡 B淋巴细胞瘤-2/B淋巴细胞瘤-2相关X蛋白通路 大鼠 

分 类 号：R285.5[医药卫生—中药学]