黄芩素对瘢痕疙瘩成纤维细胞增殖和凋亡的影响

Effect of baicalein on proliferation and apoptosis of keloid fibroblast

目的探讨黄芩素对人体瘢痕疙瘩成纤维细胞在增殖、凋亡的影响,为临床瘢痕疙瘩的治疗提供实验基础。方法 MTT法测定黄芩素干预后瘢痕疙瘩成纤维细胞的活力和乳酸脱氢酶的含量;Annexin V-FITC分析成纤维细胞在干预后的细胞凋亡率;蛋白印迹实验检测凋亡蛋白表达水平的变化。结果黄芩素对体外培养的瘢痕疙瘩成纤维细胞的增殖有抑制作用(P<0.01),抑制效应呈剂量依赖性,作用24 h后,黄芩素50μM组抑制率为65%,IC50值为(31.34±0.29)μM;黄芩素还可诱导瘢痕疙瘩成纤维细胞凋亡,黄芩素25μM组凋亡率达22.7%,和对照组相比,差异有统计学意义(P<0.01);但不影响乳酸脱氢酶的含量。黄芩素作用后明显降低线粒体膜电位,上调凋亡蛋白裂解半胱天冬酶3和裂解半胱天冬酶9的表达,裂解半胱天冬酶8的表达不受影响。结论黄芩素通过抑制细胞增殖和促进细胞凋亡来抑制瘢痕疙瘩生长。

objective To study the role of baicalein on the proliferation and apoptosis of human keloid fibroblasts and provide experimental basis for clinical treatment of keloid. Methods MTT method was applied to determine the viability and the content of lactate dehydrogenase in keloid fibroblasts after baiealein treatment; Annexin V - FITC analysis was used for detection of apoptosis of fibroblasts after baicalein treatment; Western bloting was used to detect the expression levels of fiborblasts. Results The proliferfation of keloid fibroblasts was inhibited by Baicalein in dose-dependent way and the inhibition rate of with Baicalein 50 μM was 65% 24 hours after treatment and the IC50 was （31.34±0.29）μM. Baiealein could also induce the apoptosis of keloid fibroblasts with the apoptotic rate of 22.7% ,showing significant difference compared to control group（P〈0.01）; But it did not affect the content of lactic dehydrogenase. It could reduce mitoehondria membrane potential and up-regulate the expression of cleaved easpase 3 and 9 levels and but the level of cleaved caspase 8 was not affected. Conclusion Baicalein might inhibit the growth of keloid by inhibiting proliferation and promoting apoptosis of fibroblas.