穿孔素和Fas配体在柯萨奇B3病毒性心肌炎小鼠心肌浸润细胞中表达

Expressions of perforin and Fas ligand in infiltrating cells in murine myocardium with acute myocarditis caused by coxsackievirus B3

目的探讨穿孔素（ＰＦＰ）和Ｆａｓ配体（Ｌ）介导的细胞毒作用在病毒性心肌炎发病中的作用。方法将４０只ＢＡＬＢ／ｃ小鼠随机等分为实验组和对照组，分别用柯萨奇Ｂ３病毒（ＣＶＢ３）及不含病毒的病毒稀释液经腹腔接种，于接种后７天处死，取其心脏。应用免疫组化、逆转录－多聚酶链反应（ＲＴ－ＰＣＲ）和原位杂交等方法，检测细胞介导的细胞毒作用的主要效应分子ＰＦＰ和ＦａｓＬ在心肌浸润细胞中的表达。结果（１）实验组小鼠的心肌组织中均有ＰＦＰ和ＦａｓＬ抗原阳性细胞浸润，对照组则无；（２）ＲＴ－ＰＣＲ检测发现实验组鼠的心肌组织中ＰＦＰ和ＦａｓＬｍＲＮＡ的阳性率均为１００％，明显高于对照组的２０％和３０％（Ｐ均＜０．０１）；（３）原位杂交检查显示，实验组小鼠心肌组织中均可见ＰＦＰ和ＦａｓＬｍＲＮＡ阳性的浸润细胞，而对照组则均未发现。结论ＣＶＢ３小鼠心肌炎急性期，其心肌浸润细胞中有ＰＦＰ和ＦａｓＬ表达。

To clarify the role of perforinand Fas Ligand (L)mediated cytotoxicity on pathogenesis of viral myocarditis,40 BALB/c mice were randomly divided into experimental group(n=20) and control group(n=20),and inoculated intraperitoneally with coxsackievirus B3(CVB3) and Eagles’ solution without CVB3,respectively.The mice were sacrificed and their hearts were examined on day 7 postinoculation.Expressions of perforin and FasL were detected with immunohistochemistry,reverse transcriptionpolymerase chain reaction(RTPCR) and in situ hybridization.Results:(1)Perforinand FasLpositive cells were demonstrated in experimental murine hearts by immunohistochemistry,however,no such cells were discovered in control murine hearts;(2)Test of RTPCR showed the positive ratios of perforin and FasL mRNA in myocardium were significantly higher in experimental group(100% and 100%) than those in control group(20% and 30% P<0.01);(3)Positive signals of perforin and FasL.mRNA were found in myocardium of all experimental mice by in situ hybridization,but nothing was detected in control group.The results indicated that perforin and FasL could be expressed in infiltrating cells in murine myocardium with acute myocarditis caused by coxsackievirus B3,suggesting perforin and FasL might play an important role in pathogenesis of viral myocarditis.