水蛭素对瘢痕成纤维细胞基质金属蛋白酶作用的实验研究(1)
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[摘要]目的：探讨水蛭素对体外培养的增生性瘢痕成纤维细胞基质金属蛋白酶(matrix metalloproteinases，MMPs)表达水平的影响。方法：取10例烧伤愈合后6个月内瘢痕挛缩需手术治疗的患者的瘢痕组织，采用组织块法分离培养增生性瘢痕成纤维细胞。取第4～7代细胞实验，加入0、1、10、50U/ml的水蛭素干预。观察加药后24h、48h成纤维细胞形态学变化，利用MTT(四甲基偶氮唑蓝)比色法及酶联免疫吸附试验(Enzyme-linked immunosorbent assay，ELISA)检测水蛭素对增生性瘢痕成纤维细胞(hypertrophic scar fibroblast，HSFB)及基质金属蛋白酶(MMP-2、MMP-9)的作用。结果：①1、10、50U/ml组水蛭素对HSFB均有抑制作用(P<0.05)，以50U/ml作用最明显(24h、48h抑制率分别为14.75%、15.42%)；②水蛭素作用成纤维细胞24h后 MMP-2、MMP-9表达含量均增加，分别以1U/ml、50U/ml作用最明显，差异有统计学意义(P<0.05)；48h后MMP-2表达降低,MMP-9表达有增加但无意义。结论：水蛭素能够抑制增生性瘢痕成纤维细胞增殖，能促进增生性瘢痕成纤维细胞分泌基质金属蛋白酶(MMP-2,MMP-9)有利于细胞外基质降解减少沉积。
[关键词]水蛭素；增生性瘢痕；基质金属蛋白酶(MMPs)
[中图分类号]R282.74 [文献标识码]A [文章编号]1008-6455(2012)02-0247-03
Effect of Hirudin on Matrix Metalloproteinases of Human Hypertrophic Scar Fibroblast
LI Kai-tong,LIU Da-en,LI Shun-tang,LU Hai-qiang,CHEN Xiao-ting,GAO Xing-xin
(Department of Burns and Plastic Surgery, The First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi,China)
Abstract: Objective To explore the effect of hirudin on the expression of matrix metalloproteinases of the human hypertrophic scar fibroblast. Methods 10 cases of hypertrophic scar tissue were taken from the operated patients who were within 6 months after injury . Fibroblasts of hypertrophic scar tissue were cultured by tissue block method. The cells after 4～7generations were used in experiments, which were interfered with exposure to 0U/ml,1U/ml,10U/ml or 50U/ml hirudin for 24 and 48 hours. We detected the effect of hirudin on the proliferation of fibroblasts by MTT method and the expression of MMP-2,MMP-9 by enzyme-linked immunosorbent assay (ELISA). Results ①Hirudin could inhibit the proliferation of scar fibroblasts in the group of 1,10,50U/ml(P<0.05).The effect of inhibition of hirudin with 50U/ml was most significant(the inhibition rates of 24,48hours were 14.75% and 15.42%). ②The level of expression of MMP-2 and MMP-9 was increased with the role of hirudin on fibroblasts after 24h,especially it was most significant with 1U/ml and 50U/ml ......
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