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目的研究肿瘤坏死因子样凋亡微弱诱导剂(TWEAK)影响大鼠心肌成纤维细胞(CFs)增殖和胶原合成的作用机制。方法采用胰酶消化法培养新生Wistar大鼠CFs,加入重组人TWEAK(rhTWEAK)和NF-κB抑制剂PDTC干预,qRT-PCR检测核转录因子NF-κB和金属蛋白酶9(MMP9)mRNA,Western blot检测NF-κB和MMP9蛋白表达,四甲基偶氮唑蓝(MTT)法测定细胞增殖。结果 rhTWEAK作用后NF-κB和MMP9 mRNA表达上调,同时蛋白表达明显增加,PDTC可抑制MMP9蛋白表达和细胞增殖。结论 TWEAK可通过NF-κB途径促进大鼠心肌成纤维细胞表达MMP9。
Objective To investigate the mechanism of TWEAK on the proliferation and collagen synthesis of rat cardiac fibroblasts (CFs). Methods The CFs of neonatal Wistar rats were cultured by trypsin digestion. The recombinant human TWEAK (rhTWEAK) and NF-κB inhibitor PDTC were used to intervene. The nuclear transcription factor NF-κB and MMP9 mRNA were detected by qRT-PCR. Western blot The expressions of NF-κB and MMP9 were detected, and the cell proliferation was measured by MTT assay. Results After rhTWEAK treatment, the expressions of NF-κB and MMP9 mRNA were up-regulated while the protein expression was significantly increased. PDTC inhibited MMP9 protein expression and cell proliferation. Conclusion TWEAK can promote the expression of MMP9 in rat cardiac fibroblasts through NF-κB pathway.