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目的探讨姜黄素对人胚肺成纤维细胞MRC-5在低氧下增殖和胶原表达中的作用及其机制。方法以人胚肺成纤维细胞MRC-5进行培养,随机分为常氧组(12 h,24 h,36 h)、低氧组(12 h,24 h,36 h)、低氧(12 h,24 h,36 h)+姜黄素(5μmol/L,10μmol/L,20μmol/L)组、低氧(36 h)+p38丝裂原激活蛋白激酶(p38 MAPK)阻断剂SB203580(10μmol/L)组和低氧(36 h)+三磷酸肌醇激酶/蛋白激酶B(PI3K/AKT)阻断剂LY-294002(10μmol/L)组。分别采用MTT法检测细胞增殖情况,Western blot检测MRC-5细胞中磷酸化AKT(p-AKT)、磷酸化P38(p-p38)和Ⅰ型胶原α1前体(pro-col1α1)蛋白的表达情况。结果姜黄素能抑制低氧下肺成纤维细胞的增殖,并且以低氧(36 h)+姜黄素20μmol/L组最为明显,SB203580和LY-294002能抑制MRC-5细胞增殖(P<0.05)。低氧下pro-col1α1、p-AKT、p-p38蛋白表达较常氧下增强(P<0.05),但姜黄素只对低氧下pro-col1α1、p-p38蛋白的表达有抑制,且呈剂量依赖性(P<0.05),姜黄素对p-AKT表达无明显影响(P>0.05)。结论低氧促进MRC-5细胞增殖及胶原表达可能是通过PI3K/AKT及p38 MAPK途径,姜黄素对低氧下MRC-5细胞增殖及I型胶原表达的抑制则可能是通过p38 MAPK实现。
Objective To investigate the effect of curcumin on the proliferation and collagen expression of human embryonic lung fibroblasts (MRC-5) under hypoxia and its mechanism. Methods Human embryonic lung fibroblast MRC-5 cells were randomly divided into normoxia group (12 h, 24 h, 36 h), hypoxia group (12 h, 24 h, 36 h) , 24h, 36h) + curcumin (5μmol / L, 10μmol / L, 20μmol / L) and p38 MAPK inhibitor SB203580 (10μmol / L group and LY-294002 (10μmol / L) group with hypoxia (36 h) + PI3K / AKT blocker. The proliferation of MRC-5 cells was detected by MTT assay, and the protein expressions of phosphorylated AKT, phosphorylated P38 (p-p38) and pro-col1α1 in MRC-5 cells were detected by Western blot . Results Curcumin inhibited the proliferation of lung fibroblasts under hypoxia and was most obvious in hypoxia (36 h) + curcumin (20 μmol / L) group. SB203580 and LY-294002 inhibited the proliferation of MRC-5 cells (P <0.05) . The expression of pro-col1alpha1, p-AKT and p-p38 protein in hypoxia group was higher than that in normoxia group (P <0.05), but curcumin only inhibited the expression of pro-col1alpha and p-p38 protein in hypoxia In a dose-dependent manner (P <0.05), curcumin had no significant effect on the expression of p-AKT (P> 0.05). Conclusion Hypoxia may promote the proliferation and collagen expression of MRC-5 cells through PI3K / AKT and p38 MAPK pathway. Curcumin inhibits the proliferation and expression of type I collagen in MRC-5 cells under hypoxia by p38 MAPK.