摘要
目的探讨肿瘤坏死因子样凋亡微弱诱导剂(TWEAK)影响心肌成纤维细胞(CFs)中Ⅰ型胶原和基质金属蛋白酶1(MMP-1)表达的作用机制。方法取Wistar新生大鼠CFs,加入重组人TWEAK(rhTWEAK)及P38MAPK抑制剂(SB203580)干预,将实验分为①对照组:不加干预因素;②TWEAK组:加入100 ng/mLTWEAK;③TWEAK+SB203580组:加入100 ng/mLTWEAK+SB203580。采用MTT法检测CFs增殖;Westernblot法检测Ⅰ型胶原蛋白及磷酸化P38MAPK(P-P38MAPK)蛋白表达;qRT-PCR法检测Ⅰ型胶原mRNA表达;ELISA法检测CFs细胞培养液中MMP-1的浓度。结果加入TWEAK干预后,促进了CFs增殖,Ⅰ型胶原蛋白及P-P38MAPK蛋白表达上调,Ⅰ型胶原mRNA和MMP-1表达上调。加入SB203580可抑制CFs增殖,部分抑制Ⅰ型胶原mRNA、Ⅰ型胶原蛋白、P-P38MAPK蛋白及MMP-1的表达。结论 TWEAK可通过P38MAPK途径促进CFs表达Ⅰ型胶原和MMP-1。
Objective To investigate the mechanism and effects of tumor necrosis factor-like inducer of apoptosis（TWEAK） on expressions of collagen I and matrix metalloproteinase-1（MMP-1） in cardiac fibroblasts（CFs）.Methods Cultured CFs from neonatal Wistar rats were treated with the recombinant human TWEAK and the P38MAPK specific inhibitor（SB203580）.CFs were divided into 3 groups： ① the control group： CFs were cultured without any stimulus;② the TWEAK group： CFs were incubated with 100 ng/mL TWEAK;③ the TWEAK＋SB203580 group： CFs were incubated with 100 ng/mL TWEAK＋SB203580.The CFs proliferation was examined by MTT,and expressions of collagen Ⅰ and phosphor-P38MAPK protein were detected by Western blot.qRT-PCR was used to detect the collagen Ⅰ mRNA level.ELASA was used to detect the concentration of MMP-1 in CFs cell-culture supernatant.Results TWEAK promoted the proliferation of the CFs and expressions of collagen Ⅰ and phosphor-P38MAPK protein.The expressions of collagen Ⅰ mRNA and MCP-1 were up regulated due to TWEAK.P38MAPK specific inhibitor（SB203580） could inhibit the proliferation of CFs.P38MAPK specific inhibitor（SB203580） could partially inhibit expressions of collagens Ⅰ mRNA,collagens Ⅰ protein,phosphor-P38MAPK protein and MMP-1.Conclusion TWEAK can promote expressions of collagen Ⅰ and MMP-1 in rat cardiac fibroblasts via P38MAPK pathway.
出处
《山东大学学报（医学版）》
CAS
北大核心
2012年第11期43-47,共5页
Journal of Shandong University:Health Sciences
基金
山东省科技攻关计划(2007GG10002014)
