苦参碱对瘢痕疙瘩成纤维细胞生长的抑制作用实验研究  

Inhibitory Effect of Matrine on the Growth of Keloid Fibroblasts

作　　者：徐志山 王洪一[1] 钟黎明[1] 回蔷[1] 郭冰玉[1] 陶凯[1] XU Zhi-shan;WANG Hong-yi;ZHONG Li-ming;HUI Qiang;GUO Bing-yu;TAO Kai(Department of Plastic and Reconstructive Surgery,the General Hospital of Shenyang Military Region,Shenyang 110016,Liaoning,China)

机构地区：[1]沈阳军区总医院整形外科,辽宁沈阳110016

出　　处：《中国美容医学》2018年第10期137-139,143,共4页Chinese Journal of Aesthetic Medicine

摘　　要：目的:研究苦参碱对瘢痕疙瘩成纤维细胞增殖、凋亡和自噬的影响及其相关作用机制。方法:通过收集手术切除的瘢痕疙瘩进行体外培养成纤维细胞,将细胞分为3组,分别加入10μmol/L,20μmol/L的苦参碱以及对照组(0.9%NaCl)进行实验。MTT实验检测细胞增殖情况;流式细胞技术进行细胞周期分析;Hoechest33258染色检测细胞凋亡情况;Westernblot实验检测苦参碱对瘢痕疙瘩成纤维细胞增殖、凋亡和自噬相关调控蛋白表达的影响。结果:通过不同浓度的苦参碱处理细胞后,MTT结果发现成纤维细胞的增殖受到抑制,抑制作用具有剂量依赖性;细胞周期分析发现,苦参碱作用后细胞的G1-S期转化受到了不同程度的抑制;Hoechest 33258染色发现苦参碱可以促进凋亡细胞的增加;Western blot实验结果发现苦参碱可以抑制CDK4、CDK6、Bcl-2和LC3-I蛋白的表达,促进Bax、LC3-Ⅱ和Beclin-1的表达。结论:苦参碱对瘢痕疙瘩成纤维细胞具有抑制增殖、促进凋亡的作用,能够影响细胞增殖、凋亡和自噬相关调控蛋白的表达。Objective To study the effects of Matrine on the proliferation, apoptosis and autophagy of keloid fibroblasts in vitro and its related mechanisms. Methods The fbroblasts were cultured in vitro by collecting surgical excised keloids. The cells were divided into three groups and the 10μmol/L, 20μmol/L matrine and the control group （0.9% NaCl） were added to the experiment. MTT test was used to detect the proliferation of fbroblasts, fow cytometry was used for cell cycle analysis, Hoechest 33258 staining was used to detect cell apoptosis, Western blot was used to detect the effect of Matrine on the proliferation, apoptosis and autophagy related protein expression of keloid fbroblasts. Results After treating cells with different concentrations of matrine, MTT results showed that the proliferation of fbroblasts was inhibited and the inhibitory effect was dose-dependent. The cell cycle analysis showed that the G1-S phase transformation of the cells was inhibited in varying degrees after the action of matrine. Hoechest 33258 staining showed that matrine could lead to a signifcant increase in apoptosis. The results of Western blot showed that matrine could inhibit the expression of CDK4, CDK6, Bcl-2 and LC3-I protein, and promote the expression of Bax, LC3-Ⅱ and Beclin-1. Conclusion Matrine can inhibit the proliferation and promote apoptosis of keloid fbroblasts, and can affect the cell proliferation, apoptosis and the expression of autophagy related proteins.

关 键 词：苦参碱 成纤维细胞 增殖 凋亡 自噬 细胞周期 瘢痕疙瘩 

分 类 号：R619.6[医药卫生—外科学]