紧密连接破坏及ZO-1蛋白异常表达在氯胺酮相关性膀胱炎中的意义

Significance of Tight Junction Disruption and Abnormal Expression of ZO-1 Protein in the Ketamine Associated Cystitis

目的:研究紧密连接及紧密连接蛋白ZO-1在氯胺酮相关性膀胱炎中的改变及其意义。方法:60只6-8周龄的C57小鼠随机分为氯胺酮组和对照组,2组分别按时间梯度再分为4、8、12周组,每组各10只。氯胺酮组给予氯胺酮100mg/(kg·d)腹腔注射,对照组给予等量的生理盐水腹腔注射;电镜观察膀胱组织超微结构,免疫组化分析ZO-1蛋白表达。结果:氯胺酮组4周时出现指状突起减少,细胞电子密度减低,斑点状结晶减少,紧密连接疏松。8周时细胞管腔面变平,指状突起消失,紧密连接间距离进一步增大。12周时,除上述变化外,紧密连接几乎破坏,黏膜层出现断裂,黏膜下纤维化明显。氯胺酮组从4周开始,ZO-1蛋白表达于细胞质中,部分呈斑块状表达,细胞膜中表达极少,部分标本未见表达。对照组4,8,12周异常表达所占比例分别为0/10,1/10,1/10;实验组分别为7/10,7/10,9/10;Fisher确切检验后P值分别为0.003,0.02,0.001。结论:紧密连接破坏及ZO-1蛋白的异常表达与氯胺酮导致的膀胱炎相关,尿路屏障的破坏可能导致了氯胺酮相关性膀胱炎。

Objective：To investigate the significance of tight junction disruption and abnormal expression of zonula occludens-1（ZO-1）protein in the ketamine induced cystitis.Methods：Sixty female C57BL/6mices（6-8weeks）randomly distributed into experimental group and control group,and the two groups were subdivided into 4,8,12 weeks groups according to the treatment time,with10 rats in each group.Ketamine,100mg/kg,was given as intraperitoneal injection daily,while controls mice were given injection of normal saline.The ultrastructure was observed by transmission election microscopy（TEM）and the expression of ZO-1was explored by immunohistochemistry.Results：The number of raised microplicaes,electron density and subapical vesicles were decreased after 4weeks.Tight junctions became loose.At the end of 8weeks,the surface bladderurothelium become flatten,tight junctions appeared widen,and denudation was found on epithelia.Transmission electron microscope showed that surface bladder urothelium become flatten,and tight junctions appeared widen.In addition to the above description,tight junctions was almost destroyed,while increased collagen and denuded epithelium were observed in 12 weeks.In the ketamine group,ZO-1protein was expressed in the cytoplasm in a patchy pattern,rarely outlined membrane,even it was absent.The ketamine treated group vs control group findings were abnormal in 7of 10（P=0.003）after 4weeks,7of 10（P=0.02）after 8weeks,and 9of 10（P=0.001）after 12 weeks.Conclusion：Tight junction disruption and abnormal expression of ZO-1protein associate with ketamine induced cystitis,and the changes of bladder permeabity barrier may induce the cystitis.