温肾生精饮对生精细胞凋亡和组蛋白H3K9二甲基化的影响  

Effect of “Wenshen Shengjing Decoction ” on spermatogenic cell apoptosis and histone H3K9 dimethylation

作　　者：潘晓燕[1] 王喜艳[1] 王雪楠[2] 孙占轩 王丹[1] 王晓[1] 杨胤颖 李质馨[1] 

机构地区：[1]吉林医药学院组织学与胚胎学教研室,吉林吉林132013 [2]济宁医学院附属医院生殖医学中心,山东济宁272029

出　　处：《上海中医药杂志》2014年第11期82-86,共5页Shanghai Journal of Traditional Chinese Medicine

基　　金：吉林省科技发展计划重点攻关项目(20140204033YY);吉林省中医药管理局中医药科技项目(2012-165);吉林省吉林市科技发展计划资助项目(2013625023);山东省高等学校科技计划项目(J13LL04);吉林省大学生创新创业训练计划项目

摘　　要：目的研究温肾生精饮对环磷酰胺致小鼠睾丸生精细胞凋亡和组蛋白H3K9二甲基化(H3K9me2)的影响。方法将8周龄的60只昆明种雄性小鼠随机分为对照组、模型组、西药组和中药组,每组15只。对照组腹腔注射0.9%Na Cl溶液,模型组、西药组和中药组连续5天腹腔注射环磷酰胺(80μg·g-1·d-1的)然后将对照组和模型组小鼠灌胃0.9%Na Cl溶液,西药组、中药组小鼠分别灌胃克罗米芬、温肾生精饮。各组均干预30天,检测小鼠的体质量、睾丸重和附性腺重;观察睾丸生精上皮的发育且进行Johnsen评分;TUNEL法检测睾丸生精细胞的凋亡;免疫荧光法检测睾丸生精小管中H3K9me2蛋白的表达。结果与模型组和西药组比较,中药组显著提高了生精障碍小鼠的体质量、附性腺指数和小鼠睾丸组织的Johnsen评分(P<0.05),促进了生精上皮的发育,显著减少了小鼠睾丸组织中凋亡的生精小管数和生精小管内凋亡的阳性细胞数(P<0.05),上调了精子细胞中H3K9me2的表达水平。结论温肾生精饮能显著修复环磷酰胺致小鼠睾丸生精功能损伤,其修复机制可能与减少睾丸生精细胞凋亡,稳定精子细胞中H3K9me2的表达水平有关。Objective To observe the effect of ＂Wenshen Shengjing Decoction＂ on spermatogenic cell apoptosis and histone H3K9 dimethylation. Methods Sixty mice at the age of 8 weeks were randomized into control group, model group, western medicine group and Chinese medicine group, with 15 mice in each group. Mice in the control group received intraperitoneally normal saline, and mice in the model group, western medicine group and Chinese medicine group were injected intraperitoneally with 80 μg·g-1·d-1 cyclophosphamide for five days continuously. Then mice in the control group and model group were gavaged with normal saline, and mice in the western medicine group and Chinese medicine group were garaged with clomifene citrate or ＂Wenshen Shengjing Decoction＂, with the conrse of 30 days. The body weight, testis weight and sex accessory gland weight were recorded, and the structure of seminiferous epithelium was observed and quantified with the Johnsen score. The apoptosis of spermatogenic cells in the testis tissues was detected by TUNEL assay. The expression of H3K9me2 in spermatogenic ceils was detected by immunofluorescence method. Results Compared with model group and western medicine group, the body weight growth, sex accessory gland index, development of seminiferous epithelium, and seminiferous tubule scores through the Johnsen score were significantly increased （ P 〈 0.05 ）, while the numbers of apoptotic seminiferous tubule and apoptotic spermatogenic cells were significantly decreased in Chinese medicine group （ P 〈 0.05 ）. The expression of H3K9me2 protein in spermatids was greatly improved. Conclusion These results suggest that ＂Wenshen Shengjing Decoction＂ can significantly improve the impaired reproductive function induced by cyclophospamide in mice; its mechanism may be association with reducing the spermatogenic cell apoptosis in testis and maintaining the H3K9me2 level in spermatids.

关 键 词：温肾生精饮 环磷酰胺 睾丸 细胞凋亡 组蛋白H3K9二甲基化 

分 类 号：R285.5[医药卫生—中药学]