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目的探讨体外移植整合素连接激酶(ILK)转染的骨髓间充质干细胞(BM-MSCs)抑制心肌梗死后心脏纤维化的机制。方法收集培养72 h的MSCs和ILK-MSCs后的培养基,分别与心脏成纤维细胞(CFB)共培养48 h,用5-乙炔基-2′脱氧尿嘧啶核苷(EdU)增殖试剂盒检测CFB的增殖能力,采用实时荧光定量PCR检测CFB表达金属蛋白酶(MMPs)、金属蛋白酶抑制剂(TIMPs)及细胞外基质的组成成分Ⅰ型胶原和Ⅲ型胶原的情况。结果与对照组比较,MSCs培养液组、ILK-MSCs培养液组可抑制CFB的增殖和减少Col1a1、Col3a1、TIMP-1、TIMP-2和-αSMA的基因表达(P<0.05),增加MMP-2、MMP-9的基因表达(P<0.05);与MSCs培养液组比较,ILK-MSCs的培养液组可进一步抑制Col3a1和TIMP-2的基因表达(P<0.05),并增加MMP-2的基因表达(P<0.05)。结论 ILK可能通过增加MSCs的旁分泌,抑制CFB的增殖和胶原合成能力,从而在体内发挥降低心肌梗死后纤维化的作用。
OBJECTIVE: To investigate the mechanism of BM-MSCs transfected with integrin-linked kinase (ILK) in vitro to inhibit cardiac fibrosis after myocardial infarction. Methods The media of MSCs and ILK-MSCs cultured for 72 h were collected and co-cultured with cardiac fibroblasts (CFB) for 48 h. The cells were detected by 5-ethynyl-2 ’deoxyuridine (EdU) CFB were used to detect the expression of MMPs, TIMPs and type I collagen in CFB by real-time fluorescence quantitative PCR. Results Compared with the control group, MSCs culture medium and ILK-MSCs culture medium inhibited the proliferation of CFB and decreased the gene expression of Col1a1, Col3a1, TIMP-1, TIMP-2 and -αSMA 2 and MMP-9 (P <0.05). Compared with MSCs culture medium, ILK-MSCs culture medium could inhibit the gene expression of Col3a1 and TIMP-2 (P <0.05) and increase the expression of MMP-2 Of the gene expression (P <0.05). Conclusion ILK may play a role in reducing fibrosis after myocardial infarction by increasing paracrine of MSCs, inhibiting the proliferation and collagen synthesis of CFB.