抑制c-Kit和Cx43对膀胱收缩功能的影响及其作用机制的研究  

Study of the effect of inhibition of c-Kit and Cx43 on bladder contractile function and its mechanism of action

作　　者：谭明辉 欧阳松[1] 钱彪[1] 郝雨 轩留明 孙鹏 张永强 王勤章[1] Tan Minghui;Ouyang Song;Qian Biao;Hao Yu;Xuan Liuming;Sun Peng;Zhang Yongqiang;Wang Qinzhang(Dept of Urology,The First Affiliated Hospital of Shihezi University,Shihezi 832000)

机构地区：[1]石河子大学第一附属医院泌尿外科,石河子832000

出　　处：《安徽医科大学学报》2022年第12期1941-1947,共7页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81960147)。

摘　　要：目的 通过抑制膀胱Cajal间质细胞(ICCs)上的缝隙链接蛋白43(Cx43)与酪氨酸激酶受体蛋白(c-Kit)探究对膀胱收缩功能的影响。方法 80只雄性豚鼠随机分为空白对照组、Glivec(c-Kit抑制剂)组、Gap27 (Cx43抑制剂)组和Glivec+Gap27组。4组于每日上午灌注生理盐水、Glivec、Gap 27、Glivec+Gap 27,持续灌注2个月。于灌注2个月后行尿流动力学检测造模是否成功,收取新鲜膀胱组织,行体外肌条实验检测各组的肌肉收缩功能;免疫荧光检测c-Kit与Cx43在ICCs细胞上的表达;qRT-PCR和Western blot法检测膀胱组织中c-Kit与Cx43的表达;电镜观察膀胱平滑肌超微结构变化。结果 尿动力学显示实验组较空白对照组膀胱顺应性增加(P<0.05);Glivec+Gap27组膀胱顺应性比Glivec组和Gap27组增加(P<0.01)。体外肌条实验显示实验组的膀胱肌条收缩频率和张力较空白对照组降低(P<0.05),实验组给予乙酰胆碱(ACH)后与对照组相比肌条收缩减弱(P<0.05)。免疫荧光显示c-Kit与Cx43在ICCs细胞上共表达。qRT-PCR和Western blot提示抑制c-Kit后,膀胱组织中的Cx43蛋白表达水平、基因表达水平低于空白对照组(P<0.05);抑制Cx43后,膀胱组织中的c-Kit蛋白表达水平、基因表达水平低于空白对照组(P<0.05)。电镜显示同时抑制c-Kit与Cx43后膀胱平滑肌线粒体结构破坏。结论 Cx43在膀胱ICCs上表达,二者可能共同参与调控膀胱收缩功能。Cx43和c-Kit的降低可能破坏了膀胱平滑肌的线粒体,影响其功能,进而影响膀胱收缩功能。Objective To investigation of the interaction between interstitial cells of Cajal(ICCs) and connexin 43(Cx43) in bladder contraction and its significance. Methods Eighty male guinea pigs were randomly divided into blank control group, Glivec group, Gap27 group and Glivec+ Gap27 group. Four groups of guinea pigs were perfused with saline, Glivec, Gap 27, and Glivec+Gap 27 every morning for 2 months. Success of urodynamic testing model after 2 months. Bladder tissue was collected for an in vitro muscle strip test to detect muscle contraction in each group. Correlation between c-Kit and Cx43 was detected by immunofluorescence. The interaction between c-Kit and Cx43 in the bladder was further validated by qRT-PCR and Western blot. Ultrastructural changes in the muscle layer of the bladder were observed by electron microscopy. Results Urodynamics revealed increased blad-der compliance in the experimental group compared to the blank control group(P<0.05);bladder compliance increased in the Glivec + Gap27 group compared to the Glivec and Gap27 groups( P < 0. 01). In vitro muscle strip experiments revealed that the frequency and tone of bladder muscle strip contractions were lower in the experimental group compared to the blank control group( P < 0. 05),and that muscle strip contractions were weaker in the experimental group after administration of acetylcholine( ACH) compared to the control group( P < 0. 05). Immunofluorescence showed that c-Kit was co-expressed with Cx43 on ICCs cells. qRT-PCR and Western blot suggested that the protein expression level and gene expression level of Cx43 in bladder tissues were lower after inhibition of c-Kit than in the blank control( P < 0. 05);after inhibition of Cx43,the protein expression level and gene expression level of c-Kit in bladder tissues the levels of c-Kit protein expression and gene expression in bladder tissues were lower than those in the blank control group( P < 0. 05). Electron microscopy revealed that the mitochondrial structure of bladder smooth muscle was disrupt

关 键 词：逼尿肌活动低下 CAJAL间质细胞 缝隙链接蛋白43 超微结构 豚鼠 

分 类 号：R694.5[医药卫生—泌尿科学]