TWEAK通过ERK1/2通路促进大鼠心肌成纤维细胞MMP2与Ⅰ型胶原表达

doi:10.6040/j.issn.1671-7554.0.2016.212

摘要/Abstract

摘要： 目的探讨肿瘤坏死因子样凋亡微弱诱导剂(TWEAK)调控大鼠心肌成纤维细胞(CFs)表达基质金属蛋白酶2(MMP2)及Ⅰ型胶原的作用机制。方法胰酶消化法分离培养新生大鼠原代CFs,利用Western blotting法检测磷酸化ERK1/2(p-ERK1/2)蛋白的表达水平,从而确定重组人TWEAK(rhTWEAK)及ERK1/2通路抑制剂PD98059对CFs的最佳干预浓度及干预时间。采用实时荧光定量PCR法(RT-PCR)以及Western blotting法检测干预后MMP2及Ⅰ型胶原的mRNA与蛋白表达水平,采用四甲基偶氮唑蓝(MTT)法检测不同处理对细胞增殖的影响。结果 100 μg/L TWEAK干预CFs显著上调p-ERK1/2的蛋白表达、上调MMP2及Ⅰ型胶原mRNA与蛋白的表达水平,同时显著促进细胞增殖。抑制剂PD98059阻断ERK1/2通路后显著抑制MMP2与Ⅰ型胶原的mRNA及蛋白表达,抑制CFs的增殖。结论 TWEAK通过ERK1/2通路促进CFs表达MMP2及Ⅰ型胶原。

关键词: Ⅰ型胶原, 基质金属蛋白酶2, 肿瘤坏死因子样凋亡微弱诱导剂, 细胞外信号调节激酶

Abstract: Objective To explore the mechanism of tumor necrosis factor-like weak inducer of apoptosis(TWEAK)mediating the expression of MMP2 and collagen Ⅰ in cultured neonatal rat cardiac fibroblasts(CFs). Methods CFs were isolated and cultured using trypsin enzyme digestion technique. The protein expression of phosphorylated-ERK(p-ERK1/2)was detected with Western blotting, and then the optimal interventional concentration and duration of rhTWEAK and inhibitor PD98059 on ERK1/2 pathway in CFs were determined. The protein and mRNA expressions of MMP2 and collagen Ⅰ were investigated subsequently with real-time PCR(RT-PCR)and Western blotting. The effects of different treatments on cell proliferation were assessed with methyl thiazolyl tetrazolium(MMT). Results The 100 μg/L TWEAK significantly upregulated the protein expression of p-ERK1/2, improved the expressions of MMP2 and collagen Ⅰ at the transcriptional and translational level, and promoted cell proliferation. PD98059 inhibited the protein and mRNA expressions of MMP2 and collagen Ⅰ as well as the proliferation of CFs by blocking ERK1/2 signaling pathway. Conclusion TWEAK promotes the expression of MMP2 and collagen Ⅰ in CFs via ERK1/2 signaling pathway.

Key words: Tumor necrosis factor-like weak inducer of apoptosis, Collagen Ⅰ, Matrix metalloproteinase 2, Extracellular signal-regulated kinase 1/2

中图分类号:

R542.2

巨媛媛,任满意,李睿,赵萌萌,隋树建. TWEAK通过ERK1/2通路促进大鼠心肌成纤维细胞MMP2与Ⅰ型胶原表达[J]. 山东大学学报（医学版）, 2016, 54(5): 23-28.

JU Yuanyuan, REN Manyi, LI Rui, ZHAO Mengmeng, SUI Shujian. Tumor necrosis factor-like weak inducer of apoptosis promotes expression of matrix metalloproteinase 2 and collagen Ⅰ in rat cardiac fibroblasts via the ERK1/2 signaling pathway[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2016, 54(5): 23-28.

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