朱红膏对基质金属蛋白酶活性及成纤维细胞分泌的影响  

Effects of Zhuhong ointment on MMPs activities and production by HSF

作　　者：林燕[1] 戴淼可[2] 何秀娟[3] 李萍[1] 

机构地区：[1]北京市中医研究所,北京100010 [2]北京垂杨柳医院,北京100022 [3]首都医科大学,北京100069

出　　处：《中国中药杂志》2013年第11期1795-1799,共5页China Journal of Chinese Materia Medica

基　　金：北京市自然科学基金项目(7072014);国家"十一五"科技支撑计划项目(2008BAI53B085)

摘　　要：目的:观察朱红膏是否通过对基质金属蛋白酶的影响来促进创面的愈合。方法:收集阳证疮疡创面渗液8例,检测朱红膏对渗出物中明胶酶MMP-2,MMP-9酶活性的影响;在蛋白水平观察朱红膏对MMP-1,-2活性的影响;检测朱红膏对HSF细胞的细胞毒性及IC50及用药前后HSF细胞MMP-1,-2,-9的蛋白表达。结果:朱红膏对创面渗出液中MMP-2活性有明显的抑制作用,与对照组相比P<0.01;对创面渗出液中MMP-9活性在1 g.L-1时与对照组比较无差异,但高于5 g.L-1时有明显抑制作用,与对照组比较有差异。当汞质量浓度为320 mg.L-1时,朱红膏溶液能直接抑制MMP-1活性(P<0.01)和MMP-2活性(P<0.05);但当汞质量浓度为2.56～0.51 mg.L-1时,朱红膏溶液能增强MMP-1活性(P<0.05);同时当汞在0.51～64 mg.L-1时,朱红膏溶液能够增强MMP-2活性(P<0.01)。朱红膏在DMEM的饱和浓度为39.6 mg.L-1,当汞质量浓度大于1.23 mg.L-1时有细胞毒性,HSF细胞IC50汞质量浓度为61.11 mg.L-1,在1.23,0.62,0.31 mg.L-1组均可促进HSF细胞MMP-1的蛋白表达(P<0.01);朱红膏1.23,0.62 mg.L-1组对HSF细胞MMP-2蛋白表达有抑制作用,作用不明显,0.31 mg.L-1组对MMP-2蛋白表达有促进作用,作用不明显;在汞0.62 mg.L-1时,朱红膏可促进HSF细胞表达MMP-2(P<0.01);但在这3个汞浓度条件下,朱红膏均可降低HSF细胞MMP-9的蛋白表达。结论:朱红膏在高浓度情况下对创面渗出液的MMP-2,MMP-9均有抑制作用,且抑制作用呈朱红膏含量依赖性增强;并直接抑制体外激活的MMP-1,MMP-2活性。但低浓度情况下却刺激正常成纤维细胞MMP-1,2蛋白的表达及体外酶活性;但对正常成纤维细胞MMP-9的表达表现出抑制效应。HuaFu Shengji is the primary traditional Chinese medicine（TCM） therapy for treating chronic skin ulcer. The high activities of the protein enzyme in the wound fluids is one of the main cause of healing delay.In order to investigate the effect of TCM Zhuhong ointment for promoting wound healing, This research focused on its influence on matrix metalloproteinase （MMP）activities in wound fluids with TCM Yang syndromes, directly on the activated MMP-1,2 activities in vitro and on MMP-1,-2,-9 production by HSF. 8 wound fluid samples were collected, which were diagnosed Yang Syndromes in TCM.Wound fluid activities of MMP-2 and MMP-9 were measured by gelatin zymogram assay. MMP-1 and MMP-2 activities in vitro were measured by substrate cleavage. CCK-8 was used to observe the toxicity of Zhuhong ointment on HSF. MMP-1,-2,-9 production by HSF were detected by confocal microscope. Zhuhong ointment from 1 to 25 g·L-1obviously inhibited MMP-2 activity in wound fluid.When Zhuhong ointment was over 5 g·L-1, it showed significantly inhibitory effect on wound fluid MMP-9 activity. In vitro study, when the mercury concentration was 320 mg· L-1, Zhuhong ointment solution directly inhibited both MMP-1 activity and MMP-2.But mercury concentration from 0.51-2.56 mg· L-1, it could activate MMP-1 activity, and from 0.51-64 mg·L-1, activate MMP-2 activity instead.The mercury concentration when Zhuhong ointment saturated in DMEM was 39.6 mg· L-1. When the mercury concentration was over 1.23 mg· L-1, Zhuhong ointment showed toxicity to HSF. At 1.23,0.62,0.31 mg ·L-1 of mercury concentration, it increased MMP-1 expression by HSF, and at 1.23, 0.62 mg·L-1, decreased MMP-2 expression. However,at 1.23,0.62, 0.31 mg· L-1, it decreased MMP-9 expression. At higher concentration, Zhuhong ointment can inhibit MMP-2,MMP-9 activities in wound fluid with dose-dependent way and show a direct inhibitory effect on activated MMP-1 and MMP-2 in vitro.But at a lower concentration,it showed two-way adjustment, with increased MMP-1, MMP-2 a

关 键 词：朱红膏 慢性皮肤溃疡 基质金属蛋白酶类 

分 类 号：R285[医药卫生—中药学]