川芎嗪对CVB3感染小鼠心肌细胞凋亡的影响

Effects of Ligustrazine on Cardiomyocyte Apoptosis in Mouse Infected with Coxsackie Virus B3

目的探讨川芎嗪对病毒性心肌炎(VM)小鼠心肌细胞凋亡的影响。方法将50只BALB/c小鼠随机分为A、B、C、D、E 5组,每组10只。A组腹腔注射生理盐水0.5 mL;B、C、D、E组均腹腔注射柯萨奇病毒B组(CVB3)0.5 mL诱导小鼠,建立VM模型。1 d后,A、B组给予腹腔注射生理盐水10 mg.kg-1,1次.d-1,连用7 d;C、D、E组分别腹腔注射盐酸川芎嗪注射液5 mg.kg-1、10 mg.kg-1、15 mg.kg-1,1次.d-1,均连用7 d;9 d后处死小鼠。采用TUNEL法检测心肌细胞凋亡;光镜下观察心肌病理变化;免疫组织化学技术检测各组小鼠心肌细胞中Fas/FasL蛋白的表达。结果心肌病理变化:A组无病变;心肌病变积分B组明显高于C、D、E组(均P<0.01);细胞凋亡:与A组比较,B、C、D、E组细胞凋亡指数明显升高(P<0.01或P<0.05),与B组比较C、D、E组细胞凋亡指数明显降低(P<0.01);Fas/FasL蛋白阳性染色指数:B、C、D、E组均显著高于A组(P<0.01或P<0.05),C、D、E组显著低于B组(P<0.01)。结论川芎嗪可通过下调病毒性心肌炎小鼠心肌细胞Fas/FasL蛋白表达,减少心肌细胞凋亡和心肌损伤。

Objective To investigate the effects of Ligustrazine on cardiomyocyte apoptosis in mouse with viral myocarditis.Methods A total of 50 BALB/c mice were randomly divided into 5 groups.Mice in group A were inoculated with 0.5 mL normal saline by peritoneal injection;Mice in group B,C,D,E inoculated with the Coxsackie Virus B（CVB3）by peritoneal injection were adopted as viral myocarditis model.One day later,Mice in group A and B were given 10 mg·kg^-1 normal saline by peritoneal injection for 7 days;Mice in group C,D and E were given 5 mg·kg^-1,10 mg·kg^-1,15 mg·kg^-1 Ligustrazine by peritoneal injection for 7 days.All the mice were sacrificed on the 9th day after modeling.The cardiomyocyte apoptosis was detected by TDT/Dutp nick end labeling（TUNEL）.The myocardial pathologic changes were observed,and the expression of Fas/FasL protein in myocardium was determined by immunohistochemistry.Results The myocardial pathologic change was not found in group A.Pathology scores in group B was significantly increased than that in group C,D and E（P〈0.01）;Apoptosis index in the group B,C,D,E were higher than that of group A（P〈0.01 or P〈0.05）,Apoptosis index in the group C,D,E were significantly lower than that in group B（P〈0.01）;Levels of Fas/FasL expression in he group B,C,D,E were higher than that of group A（P〈0.01 or P〈0.05）,Levels of Fas/FasL expression the group C,D,E were significantly lower than that in group B（P〈0.01）.Conclusion The expression of Fas/FasL protein may be down-regulated in the viral myocarditis in mice treated with Ligustrazine.The change may contribute to reduce the apoptosis of cardiomyocyte and myocardial injury.