摘要
目的探讨氧化苦参碱对人增生性瘢痕成纤维细胞增殖及凋亡的影响。方法从人的增生性瘢痕组织中提取成纤维细胞进行培养,通过MTT实验观察不同浓度的氧化苦参碱对成纤维细胞增殖的影响。应用流式细胞术检测经氧化苦参碱处理后成纤维细胞的周期变化,并通过Annexin V-FITC/PI双染色法和Hoechst 33258染色观察细胞的凋亡情况。通过Western Blot和RT-PCR检测氧化苦参碱对细胞周期蛋白CDK4、CDK6,以及凋亡调节蛋白Bcl-2和Bax的作用。结果与对照组相比,氧化苦参碱的浓度为10μmol·L^(-1)时,24 h细胞增殖的抑制率为(18.92±3.77)%,成纤维细胞的早期和晚期凋亡率分别为(3.28±2.56)%和(2.98±1.78)%;氧化苦参碱的浓度为20μmol·L^(-1)时,24 h细胞增殖的抑制率为(32.93±2.68)%,成纤维细胞的早期和晚期凋亡率分别为(6.89±0.76)%和(1.47±2.57)%。氧化苦参碱能够通过抑制细胞周期蛋白CDK4、CDK6和抗凋亡蛋白Bcl-2的表达,诱导促凋亡蛋白Bax的表达来影响细胞周期进程,将细胞周期阻滞于G1期,使细胞增殖受到抑制,凋亡率增加。结论氧化苦参碱能够抑制人增生性瘢痕成纤维细胞的增殖并诱导其凋亡,可作为预防和治疗增生性瘢痕的潜在药物。
Objective To explore the effect of oxymatrine on proliferation and apoptosis of human hypertrophic scar fibroblasts.Methods Fibroblasts derived from human hypertrophic scar were isolated and cultured.The effect of different concentrations of oxymatrine on proliferation of fibroblasts was detected by MTT assay.After oxymatrine treated,the change of fibroblast cycle was examined by flow cytometry.The condition of fibroblasts apoptosis was observed through Annexin V-FITC/PI and Hochest 33258 staining.Effect of oxymatrine on cell cyclin protein of CDK4 and CDK6,and apoptosis-related protein of Bcl-2 and Bax was verified by Western Blot and RT-PCR assay.Results Compared with the control group,when oxymatrine concentration was 10μmol·L-1,the inhibitory rate of 24 h cell proliferation was(18.92±3.77)%,and the early and late apoptosis rates were(3.28±2.56)%and(2.98±1.78)%,respectively.When the concentration of oxymatrine was 20μmol·L-1,the inhibitory rate of 24 h cell proliferation was(32.93±2.68)%,and the early and late apoptosis rates were(6.89±0.76)%and(1.47±2.57)%,respectively.Oxymatrine can affect cell cycle progression by inhibiting the expression of cell cyclin protein of CDK4 and CDK6 and promoting the expression of apoptotic protein of Bcl-2 and Bax.Cell cycle was blocked in G1 phase and cell proliferation was inhibited.The apoptosis rate was increased.Conclusion Oxymatrine can inhibit the proliferation and promote the apoptosis of human hypertrophic scar fibroblasts,and it can be served as a potential drug for the prevention and treatment of hypertrophic scar.
作者
回蔷
郭冰玉
张倩
常鹏
徐志山
陶凯
HUI Qiang;GUO Bingyu;ZHANG Qian;CHANG Peng;XU Zhishan;TAO Kai(Department of Plastic Surgery,the General Hospital of North Theater,Shenyang 110840,China)
出处
《沈阳药科大学学报》
CAS
CSCD
北大核心
2019年第5期415-420,共6页
Journal of Shenyang Pharmaceutical University
基金
辽宁省自然科学基金资助项目(20180550992)
