Toll样受体3信号通路介导自身免疫性心肌炎炎症反应及作用机制

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目的探讨自身免疫性心肌炎大鼠心肌组织Toll样受体3(TLR3)表达水平及临床意义。方法将54只大鼠按照随机数字表法分为AM组、干预组和对照组,每组18只,对照组大鼠正常喂养,AM组、干预组建立自身免疫性心肌炎大鼠模型,干预组于模型建立第21天注射0.1 mg TLR3配体溶液,AM组、对照组大鼠注射等量PBS溶液。分别于模型建立第7、14、21、24、28、35天处死各组3只大鼠,观察各组大鼠心肌组织病理变化,酶联免疫吸附试验检测血清心肌肌凝蛋白自身抗体滴度,免疫组织化学染色法检测心肌组织TLR3蛋白表达,实时定量PCR检测心肌组织TLR3、TNF-αm RNA表达。结果对照组大鼠血清心肌肌凝蛋白自身抗体在不同时间点比较差异无统计学意义(P>0.05),干预组血清心肌肌凝蛋白自身抗体滴度在第28天达高峰,AM血清心肌肌凝蛋白自身抗体滴度在第21天达高峰;第14、21、24、28、35天AM组、干预组血清心肌肌凝蛋白自身抗体滴度显著高于对照组(P<0.05),第24、28、35天干预组血清心肌肌凝蛋白自身抗体滴度显著高于AM组(P<0.05)。对照组TLR3蛋白表达在不同时间点比较差异无统计学意义(P>0.05);第14、21、24、28、35天,AM组、干预组TLR3蛋白表达显著高于对照组(P<0.05),第24、28、35天,干预组TLR3蛋白表达显著高于AM组(P<0.05)。对照组TLR3、TNF-αm RNA相对表达量在不同时间点比较差异无统计学意义(P>0.05);第7、14、21、24、28、35天,AM组、干预组TLR3、TNF-αm RNA相对表达量显著高于对照组(P<0.05),第24、28、35天,干预组TLR3、TNF-αm RNA相对表达量显著高于AM组(P<0.05)。结论自身免疫性心肌炎心肌组织TLR3表达上调,TLR3信号通路介导的炎症反应参与了自身免疫性心肌炎的发病。 Objective To investigate the expression of Toll-like receptor 3 (TLR3) in myocardium and its clinical significance in rats with autoimmune myocarditis. Methods Fifty-four rats were randomly divided into AM group, intervention group and control group, with 18 rats in each group. Rats in control group were fed normal rats. AM rats and rats in intervention group were given autoimmune myocarditis model. At the 21st day after the model was established, 0.1 mg TLR3 ligand solution was injected, and AM rats and control rats were injected with equal volume of PBS solution. Three rats in each group were sacrificed on the 7th, 14th, 21st, 24th, 28th and 35th day after the establishment of the model. The pathological changes of myocardium in each group were observed, and the serum cardiac myosin autoantibody titer was detected by enzyme-linked immunosorbent assay The expression of TLR3 protein in myocardium was detected by immunohistochemical staining. The expression of TLR3 and TNF-αmRNA in myocardium was detected by real-time quantitative PCR. Results There was no significant difference in serum cardiac myosin autoantibodies in control group at different time points (P> 0.05). The titer of serum cardiac myosin peaked at the 28th day in the intervention group, The titers of autoantibodies to coagulation protein peaked on the 21st day. The titers of serum cardiac myosin antibodies in the intervention group were significantly higher than those in the control group on the 14th, 21st, 24th, 28th and 35th days (P <0.05) The titers of serum cardiac myosin autoantibodies in 24, 28, 35 days intervention group were significantly higher than those in AM group (P <0.05). The TLR3 protein expression in control group was not significantly different at different time points (P> 0.05). On the 14th, 21st, 24th, 28th and 35th days, TLR3 protein expression in AM group and intervention group was significantly higher than that in control group ). On the 24th, 28th and 35th days, the expression of TLR3 protein in intervention group was significantly higher than that in AM group (P <0.05). The relative expression levels of TLR3 and TNF-αmRNA in the control group had no significant difference at different time points (P> 0.05). On the 7th, 14th, 21st, 24th, 28th and 35th days, the TLR3 and TNF- (P <0.05). On the 24th, 28th and 35th days, the relative expression of TLR3 and TNF-αmRNA in the intervention group was significantly higher than that in the AM group (P <0.05). Conclusion The expression of TLR3 in myocardium of autoimmune myocarditis is up-regulated, and the inflammatory response mediated by TLR3 signaling pathway is involved in the pathogenesis of autoimmune myocarditis.
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