苦杏仁甙抑制成人肾脏成纤维细胞的增殖

Inhibitory effect of amygdalin on human renal fibroblast proliferation

背景:苦杏仁甙在体外具有抑制纤维细胞增殖的作用,而肾脏移植后一定时间内的肾活检均提示有程度不同的纤维化,苦杏仁甙是否能抑制肾脏移植后的纤维化是一个有临床意义的课题。目的:观察苦杏仁甙对成人肾脏成纤维细胞增殖的影响。设计、时间及地点:前瞻性随机对照实验,于2003-06/2007-12在南京军区福州总医院实验科完成,实验室为南京军区重点实验室。材料:苦杏仁甙经中国药品生物制品检定所鉴定,批号020903。方法:采用消化法原代培养人肾成纤维细胞,并以免疫细胞化学方法鉴定。采用MTT比色法检测含25,50,80,100,200mg/L苦杏仁甙的培养液对人肾成纤维细胞增殖的影响,以不含苦杏仁甙的培养液为对照。主要观察指标:苦杏仁甙培养液对肾脏成纤维细胞的增殖活性的影响。结果:分离出的成纤维细胞免疫细胞化学方法进行鉴定显示,vimentin呈阳性表达,keramin和desmin呈阴性表达,确认为成纤维细胞。随着培养时间的延长,对照组肾脏成纤维细胞明显增殖。不同质量浓度苦杏仁甙组肾脏成纤维细胞增殖明显低于对照组。不同质量浓度苦杏仁甙对成纤维细胞的增殖活性均有抑制作用,且呈一定剂量依赖性,以100mg/L苦杏仁甙抑制作用最强。结论:苦杏仁甙能明显抑制人肾成纤维细胞增殖,并呈剂量依赖性。

BACKGROUND： Amygdalin can inhibit fibroblast proliferation in vitro. Various degrees of fibrosis are found in renal biopsy at certain time after renal transplantation. Therefore, whether amygdalin can inhibit fibrosis following renal transplantation is of clinical significance. OBJECTIVE： To explore the effect of amygdalin on the proliferation of human renal fibroblasts. DESIGN, TIME AND SETTING： Prospective randomized controlled trial was performed at Key Laboratory of Fuzhou General Hospital of Nanjing Military Area Command of Chinese PLA between June 2003 and December 2007. MATERIALS： Amygdalin is identified by the National Institute for the Control of Pharmaceutical and Biological Products （No. 020903）. METHODS： Human renal fibroblasts isolated from human renal papilla were cultured and identified by immunocytochemistry. The influence of culture solution containing amygdalin （25, 50, 80, 100 and 200 mg/L） on the proliferation of human renal fibroblasts was detected by MTT colorimetric assay. Cells cultured in culture solution with no amygdalin served as controls. MAIN OUTCOME MEASURES： The influence of amygdalin on the proliferation of human renal fibroblasts. RESULTS： Immunocytochemistry showed that the cells were vimentin-positive and keramin and desmin-negative, indicating the cells were fibroblasts. The renal fibroblasts in control condition significantly proliferated with culture time. The cell proliferation in each amygdalin group was slow than in control group. Amygdalin inhibited human renal fibroblasts proliferation in a dose-dependent manner in certain concentration range, and 100 mg/L had amygdalin the strongest inhibition. CONCLUSION： Amygdalin could markedly inhibit human renal fibroblast proliferation in a dose-dependent manner.