电针对干眼症兔结膜细胞凋亡及相关蛋白Caspase-3、Fas和Bcl-2表达的影响

目的:通过观察电针对干眼症兔结膜细胞凋亡及相关蛋白Caspase-3、Fas和Bcl-2表达的影响,从细胞凋亡角度探讨电针治疗干眼症的作用机制.方法:雄性新西兰兔随机分为正常组、模型组、电针组和假电针组.采用0.1%苯扎氯铵滴眼制作兔干眼症模型.检测各组兔泪液分泌量、泪膜破裂时间;采用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记测定(TUNEL)法检测结膜细胞凋亡情况;应用免疫组化法检测结膜细胞Caspase-3、Fas和Bcl-2蛋白表达.结果:与正常组比较,模型组兔泪液分泌量降低,泪膜破裂时间缩短(均P<0.01);与模型组、假电针组比较,电针组兔泪液分泌量增多,泪膜破裂时间延长(均P<0.05).与正常组比较,模型组兔结膜细胞凋亡增加(P<0.01),Caspase-3和Fas蛋白表达增加(均P<0.05),Bcl-2蛋白表达降低(P<0.01);与模型组、假电针组比较,电针组兔结膜细胞凋亡减少(均P<0.01),Caspase-3和Fas蛋白表达减少(均P<0.05),Bcl-2蛋白表达增加(均P<0.01).结论:电针能抑制干眼症兔结膜细胞凋亡,下调凋亡相关蛋白Caspase-3和Fas表达,上调Bcl-2蛋白表达,该作用可能是电针治疗干眼症的作用机制之一.

Effects of electroacupuncture on conjunctival cell apoptosis and the expressions of apoptosis-related proteins Caspase-3,Fas and Bcl-2 in rabbits with dry eye syndrome

Objective:By observing the effects of electroacupuncture(EA)on the apoptosis of conjunctival cells of rabbits with dry eye syndrome(DES)and the expressions of apoptosis-related proteins Caspase-3,Fas and Bcl-2,to discuss the mechanism of EA in the treatment of DES from the perspective of cell apoptosis.Methods:Male New Zealand rabbits were randomly divided into a normal group(NG),a model group(MG),an EA group(EAG)and a sham EA group(SEAG).DES rabbit model was developed by eye drop of 0.1%benzalkonium chloride.The rabbit tear secretion and tear film break-up time(BUT)were measured;terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL)assay was used to detect the apoptosis of conjunctival cells;the expressions of Caspase-3,Fas and Bcl-2 proteins in conjunctival cells were detected by immunohistochemistry.Results:Compared with the NG,the rabbit tear secretion decreased and the BUT was shortened in the MG(both P<0.01);compared with the MG and the SEAG,the rabbit tear secretion increased and the BUT was prolonged in the EAG(all P<0.05).Compared with the NG,the apoptosis of rabbit conjunctival cells increased(P<0.01),the expressions of Caspase-3 and Fas proteins increased(both P<0.05),and the expression of Bcl-2 protein decreased(P<0.01)in the MG;compared with the MG and the SEAG,the apoptosis of rabbit conjunctival cells decreased(both P<0.01),the expressions of Caspase-3 and Fas proteins decreased(all P<0.05),and the expression of Bcl-2 protein increased(both P<0.01)in the EAG.Conclusion:EA can inhibit the apoptosis of rabbit conjunctival cells,down-regulate the expressions of apoptosis-related proteins Caspase-3 and Fas,and up-regulate the expression of Bcl-2 protein,which may be one of the mechanisms of EA in treatment of DES.