知柏地黄汤对解脲支原体感染大鼠生精细胞TRPV1、TRPV5表达的影响  

Effect of Zhibai Dihuang Decoction on Expressions of TRPV1 and TRPV5 in Spermatogenic Cells of UU-infected Rats

作　　者：何清湖[1] 郭炫佐[2] 郭军华[2] 李迎秋[3] 刘朝圣[1] 

机构地区：[1]湖南中医药大学中西医结合临床重点学科,长沙410007 [2]湖南中医药大学研究生学院,长沙410007 [3]湖南中医药大学中医学院,长沙410007

出　　处：《中国中西医结合杂志》2015年第10期1218-1222,共5页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金资助项目(No.81373641);湖南省科技厅课题资助项目(No.2011sk3104);长沙市科技局课题资助项目(No.K1301014-31);湖南省科技计划项目(No.2012SK3138)

摘　　要：目的观察知柏地黄汤对解脲支原体（Ureaplasma urealyticum,UU）感染大鼠精液及生精细胞瞬时受体电位香草酸亚型（transient receptor potential family vanilloid subtype,TRPV）1、TRPV5表达的影响,探讨UU感染导致不育症的病理机制及知柏地黄汤的干预作用。方法 4～5月龄SD大鼠60只,随机抽取45只,经膀胱接种UU悬液,建立UU睾丸感染动物模型。剩余15只同步注射生理盐水作为正常对照组,UU感染模型大鼠再随机分成模型对照组、阿奇霉素组、知柏地黄汤组,每组15只。知柏地黄汤组给予知柏地黄汤1g/（kg·d）灌胃,阿奇霉素组给阿奇霉素混悬液0.105 g/（kg·d）灌胃,正常对照组、模型对照组给予等剂量生理盐水灌胃。各组给药均每日1次,连续给药21天。处死大鼠后取睾丸、附睾组织,比较各组UU阳性率;采用机械分离法分离精子细胞,应用彩色精子动态检测系统进行精子活力运动参数测定;采用实时定量PCR法测定生精细胞TRPV1、TRPV5 m RNA表达;采用VVestern blot法检测生精细胞TRPV1、TRPV5蛋白表达。结果模型对照组UU阳性率[86.7（13门5）]明显高于正常对照组（0.0%）（P〈0.05）,而知柏地黄汤组[33.3%（5/15）]及阿奇霉素组[26.7%（4/15）]均较模型对照组降低（P〈0.05）。与正常对照组比较,模型对照组大鼠a及b级精子减少,直线速度、平均速度降低,生精细胞TRPV1、TPRV5 mRNA和蛋白表达下降,差异有统计学意义（P〈0.01）。与模型对照组比较,知柏地黄汤组和阿奇霉素组a及b级精子增多,直线速度、曲线速度及平均速度提高,TRPV1、TRPV5 mRNA和蛋白表达水平升高,差异有统计学意义（P〈0.05,P〈0.01）。与阿奇霉素组比较,知柏地黄汤组a及b级精子增多,曲线速度、平均速度提高,TRPV1、TRPV5mRNA和蛋白表达水平升高,差异有统计学意义（P〈0.05,P〈0.01）。结论知柏地黄汤具有抗UU感染、提高大鼠精液质量作用,可能与Objective To observe the effect of Zhibai Dihuang Decoction（ZDD） on mRNA and protein expressions of transient receptor potential family vanilloid subtype 1（TRPV1） and transient receptor potential family vanilloid subtype 5（TRPV5） in Ureaplasma urealyticum（UU）-infected rat semens and spermatogenic cells,and to explore the pathomechanism of UU-infected infertility and the intervention of ZDD.Methods Totally 45 were randomly selected from 60 4-5 months old SD rats.UU testicular infected animal models were set up after bladder inoculation of UU suspension.The remaining 15 rats were simultaneously injected with normal saline as a normal control group.After a successful modeling,UU infected model rats were randomly divided into the model group,the azithromycin group,and the ZDD group,15 in each group.Rats in the ZDD group were administered with ZDD at the daily dose of 1 g/kg by gastrogavage.Rats in the azithromycin group were administered with azithromycin suspension at the daily dose of 0.105 mg/kg by gastrogavage.Equal volume of normal saline was administered to rats in the normal control group and the model group.All medication was performed once daily for 21 successive days.Testes and epididymis were extracted after rats were killed and UU positive rates were compared among all groups.Sperm cells were separated using a mechanical separation technique.Sperm motility parameters were detected using color sperm motion detection system.mRNA and protein expressions of TRPV1 and TRPV5 in spermatogenic cells were determined by real-time quantitative PCR and Western blot.Results The UU positive rate was obviously higher in the model group than in the normal control group[（86.7%（13/15 cases） vs.0]P0.05）.It was lower in the ZDD group[33.3%（5/15 cases）]and the azithromycin group[26.7%（4/15 cases）]than in the model group（P0.05）.Compared with the normal control group,class A and B sperms were reduced,the linear velocity and the average velocity were significantly lowered,mRNA and protein expr

关 键 词：知柏地黄汤 解脲支原体 生精细胞 瞬时受体电位香草酸亚型1 瞬时受体电位香草酸亚型5 

分 类 号：R285.5[医药卫生—中药学]