动态观察胱天蛋白酶募集域蛋白9在心肌梗死后表达的基础研究

Dynamic observation on the expression of caspase recruitment domain protein 9 protein in post myocardial infarction

目的:探讨心肌梗死后小鼠胱天蛋白酶募集域蛋白9(caspase recruitment domain-containingprotein 9,CARD9)的表达及其与炎症的关系。方法:选择野生型小鼠和CARD9敲除小鼠各26只,分为野生型对照组、野生型心肌梗死组、CARD9敲除对照组和CARD9敲除心肌梗死组共4组,每组13只。心肌梗死模型组是通过结扎冠状动脉前降支复制心肌梗死模型,而对照组仅穿线不结扎。于模型复制后第1天,从每组随机抽取3只小鼠处死并行氯化三苯四氮唑(TTC)染色,观察模型是否复制成功;于第3天和第28天,每组分别随机各抽取3只小鼠,处死后取心肌组织行石蜡切片,苏木素伊红(HE)染色和免疫组化染色观察炎症细胞浸润,免疫组化染色和蛋白定量观察CARD9表达。结果:TTC证实心肌梗死模型复制成功;免疫组化染色和蛋白印迹证实,与野生型对照组比较,野生型心肌梗死组3d及28d时梗死区域CARD9表达均增高(P<0.05)与野生型心肌梗死组比较,3 d时CARD9敲除组心肌梗死组巨噬细胞浸润增加(P<0.05)。结论:心肌梗死后CARD9表达上调,而CARD9敲除炎症细胞浸润增加,提示CARD9参与心肌梗死并对早期炎症可能有一定抑制作用。

Objective：To explore the expression and relationship between inflammation and caspase recruitment domain protein 9（CARD9） in mice after myocardial infarction（MI）.Methods：26 WT mice and 26 CARD9 knock out（CARD9^-/-） mice were divided into the control WT group,the control CARD9^-/-mice group,the MI WT group,the MI CARD9^-/-mice group randomly.The anterior descending branch of coronary artery was ligated to establish the acute myocardium infarction model in MI group,while being thread without ligation was operated on the control mice.On the first day,one three mice were randomly selected from every group,sacrificed and stained with TTC to document MI procedure was successful.Similarly,on day 3 and 28,three WT mice were randomly selected from every group respectively.The heart sections were undergone HE staining and immunohistochemistrial staining.Immunohistochemistry staining and western-blot were performed to determine the expression of CARD9.Results：TTC staining showed that MI remodeling was established successfully.For the WT mice on day 3 and even 28,immunohistochemistry staining and Western-blot showed that the expression of the CARD9 in the infarction area was higher in the myocardial infarction group than that in the control group（P 0.05）;Compared with the myocardial infarction group of the WT mice on day 3,macrophage infiltration in the myocardial infarction group of the CARD9^-/-mice on day 3 was significantly increased（P 0.05）.Conclusion：The expression of the protein CARD9 was increased in post myocardial infarction,inflammation cell infiltration was increased in CARD9^-/-mice,it involved in the process of myocardial infarction and may inhibit early stages of inflammation.