曲尼司特对高糖培养心肌成纤维细胞增殖及表型转化的影响

摘要

OBJECTIVE To investigate the effect of tranilast on cardiac fibroblasts proliferation and phenotype transformation incubated with a high concentration of glucose and the possible mechanism. METHODS The cardiac fibroblasts were divided into seven groups in accordance with different nutrient solutions:normal control group(5.5 mmol · L-1 glucose),hypertonic group(5.5 mmol · L-1 glucose+mannitol 25 mmol · L- 1),high glucose group (25 mmol · L- 1 glucose),tranilast intervention groups (25 mmol·L-1 glucose+tranilast 50,100 and 200μmol·L-1),and activin receptor-like kinase 7(ALK7) inhibitor group(25 mmol · L-1 glucose+10μmol · L-1 SB431542). The cell proliferation was detected by MTT method. The transformation of cardiac fibroblasts was determined by immunfluorescence staining. The expression of fibroblast specific protein 1 (FSP-1),α-smooth muscle actin (α-SMA),transforming growth factor-β1(TGF-β1)and ALK7 was assessed by Western blotting. RESULTS Compared with nor⁃mal control group,A492 nm of cells in high glucose group was significantly increased(P<0.01),while the expression of α-SMA,TGF-β1 and ALK7 protein in high glucose group was significantly increased(P<0.01),but FSP-1 protein was significantly decreased(P<0.01). There was no difference between normal control group and hypertonic group. Compared with high glucose group,A492 nm of cells in tranilast or SB431542 intervention groups were decreased(P<0.05),and the expression of α-SMA,TGF-β1 and ALK7 protein was significantly decreased(P<0.05),but the expression of FSP-1 protein was increased (P<0.05)in tranilast or SB431542 intervention groups. CONCLUSION Tranilast can inhibit the proliferation and phenotype transformation of cardiac fibroblasts induced by high glucose,which may be related to down-regulation of the expression of ALK7.%目的：探讨曲尼司特对高糖培养大鼠心肌成纤维细胞增殖及表型转化的作用及可能机制。方法体外培养大鼠心肌成纤维细胞（CF）分为正常对照组（5.5 mmol·L-1葡萄糖）、高渗对照组（葡萄糖5.5 mmol·L-1+甘露醇25 mmol·L-1）、高糖组（25 mmol·L-1葡萄糖）、曲尼司特干预组（葡萄糖25 mmol·L-1+曲尼司特50，100和200μmol·L-1）及活化素受体样激酶7（ALK7）阻断剂组（葡萄糖25 mmol·L-1+SB43154210μmol·L-1）。各组细胞分别与不同药物孵育48 h后，采用MTT法检测CF存活，免疫荧光法检测CF表型转化，Western蛋白印迹法检测CF特异性蛋白1（FSP-1）、α平滑肌动蛋白（α-SMA）、转化生长因子β1（TGF-β1）和ALK7的蛋白表达。结果与正常对照组比较，高糖组CF A492 nm明显升高（P<0.01），FSP-1表达明显减少（P<0.01），α-SMA表达明显增多（P<0.01），TGF-β1和ALK7表达增多（P<0.01），而高渗对照组以上指标均无显著性差异。与高糖组比较，应用ALK7阻断剂SB431542及不同浓度曲尼司特同步干预后，均可使CF A492 nm显著降低（P<0.05）， FSP-1表达明显增多（P<0.05），α-SMA表达明显减少（P<0.01），TGF-β1表达明显减少（P<0.05），ALK7表达减少（P<0.05）。结论曲尼司特可抑制高糖诱导的CF增殖及表型转化，其机制可能与下调ALK7的表达有关。