摘要
目的:探讨五子衍宗方对环磷酰胺致成年雄性小鼠睾丸生殖细胞凋亡的保护作用。方法:随机将50只成年雄性Balb/C小鼠分为正常对照组、模型组以及五子衍宗方低(100 mg/kg)、中(200 mg/kg)、高(400 mg/kg)剂量组,每组10只。五子衍宗方3个剂量组每天分别灌胃相应剂量五子衍宗方,正常对照组和模型组灌胃等体积双蒸水,1次/d。正常对照组从第4天开始腹腔注射生理盐水,其余各组从第4天开始腹腔注射环磷酰胺200 mg/kg,每隔7 d注射1次,共4次,同时各组每天继续灌胃相应药物。第4次腹腔注射环磷酰胺后间隔12 h称量小鼠体质量,乙醚麻醉颈椎脱臼处死小鼠,迅速取出睾丸去除多余组织,称量睾丸质量,计算睾丸指数,HE染色观察睾丸组织形态变化,TUNEL检测睾丸生殖细胞凋亡,Western blot检测睾丸组织中凋亡相关蛋白Caspase-3、BAX、BCL-2表达。结果:与正常对照组比较,模型组小鼠体质量、睾丸质量、睾丸指数、睾丸内各级生殖细胞数量及睾丸组织内BCL-2蛋白表达水平显著降低,睾丸内凋亡阳性染色细胞显著增多,睾丸组织内BAX、Caspase-3蛋白表达水平显著升高;而五子衍宗方可显著升高小鼠体质量、睾丸质量、睾丸指数、睾丸内各级生殖细胞数量以及睾丸内BCL-2蛋白表达水平,明显减少睾丸内凋亡阳性染色细胞,降低睾丸内BAX和Caspase-3蛋白表达水平。结论:五子衍宗方能显著减少环磷酰胺致成年雄性小鼠睾丸生殖细胞凋亡,其作用机制可能与升高BCL-2蛋白表达水平、降低BAX、Caspase-3蛋白表达有关。
Objective: To investigate the protective effect and mechanism of Wuzi Yanzong prescription on apoptosis in germ cell of adult male mice induced by cyclophosphamide( CTX). Methods: Male Balb / C mice were randomly divided into normal control group,model group,the low-,medium- and high- dose of Wuzi Yanzong prescription groups( 100 mg / kg,200 mg / kg and 400 mg / kg),with 10 mice in each group. The mice were injected intraperitoneally with CTX( 200 mg / kg) from 4th day,and gave drug once a week,and executed for 4 weeks. Three doses of Wuzi Yanzong prescription were intragastrically administered every day. For normal control group,the same procedure was performed with intraperitoneal normal saline. Twelve hours after giving CTX at last time,all mice were weighed and sacrificed by cervical dislocation. The testis was immediately dissected and weighed,and then calculated the testis index. The pathological changes of testis were observed by HE staining,the apoptosis of germ cells were detected by TUNEL,the expression of apoptosis-related protein Caspase-3,BAX,BCL-2 in testis were examined by Western blot. Results: Compared with normal control group,the body weight,testis weight,testis index,and the expression of BCL-2 protein levels in testis of model control group were significantly decreased,the expression of BAX,Caspase-3 protein levels and apoptosis in testis of model control group were significantly increased. Wuzi Yanzong prescription significantly increased the body weight,testis weight,testis index,the expression of BCL-2 protein,while decreased the levels of BAX and Caspase-3 protein expression,and then led to the reduction in apoptosis of testis. Conclusion:Wuzi Yanzong prescription can effectively protect the apoptosis of germ cell induced by CTX,and its mechanism may be associated with downregulating protein expression of BAX and Caspase-3,and increasing the protein expression of BCL-2.
出处
《中药材》
CAS
CSCD
北大核心
2016年第5期1143-1147,共5页
Journal of Chinese Medicinal Materials
基金
国家自然科学基金(81173375
81373881
81573931
81503334)
三峡大学人才科研启动基金(KJ2014B067)
关键词
五子衍宗方
环磷酰胺
生殖细胞
细胞凋亡
Wuzi yanzong prescription
Cyclophosphamide
Germ cell
Cell apoptosis