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  

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

作　　者：巨媛媛 任满意[2] 李睿[1] 赵萌萌[1] 隋树建[1] 

机构地区：[1]山东大学第二医院心内科,山东济南250033 [2]山东大学附属千佛山医院心内科,山东济南250014

出　　处：《山东大学学报（医学版）》2016年第5期23-28,共6页Journal of Shandong University:Health Sciences

基　　金：山东省科技攻关项目(2013GSF12110)

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(rh TWEAK)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 100μg/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。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 rh TWEAK 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 The100 μg / L TWEAK significantly upregulated the protein expression of p-ERK1 /2,improved the expressions of MMP2 and collagenⅠat the transcriptional and translational level,and promoted cell proliferation. PD98059 inhibited the protein and mRNA expressions of MMP2 and collagenⅠas well as the proliferation of CFs by blocking ERK1 /2 signaling pathway.Conclusion TWEAK promotes the expression of MMP2 and collagenⅠin CFs via ERK1 /2 signaling pathway.

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

分 类 号：R542.2[医药卫生—心血管疾病]