三组中药复方对大鼠肾小球系膜细胞MMP-3/TIMP-1表达影响的实验研究

【作者】 丁晓欢；

【导师】 张君；

【作者基本信息】 辽宁中医药大学 ， 中西医结合临床， 2010， 硕士

【摘要】 目的:从基因水平研究三组中药复方对肾小球系膜细胞基质金属蛋白酶(matrix metallo proteinases, MMPS)MMP-3及基质金属蛋白酶抑制物(tissue inhibitor of metallo proteinases,TIMPS)TIMP-1mRNA表达的影响,对具有延缓和防治肾小球硬化进程的有效药味探讨其作用机理。材料与方法:1.应用血清药理学方法,制取三组中药小复方及正常组的药理血清。2.采用体外培养法培养大鼠肾小球系膜细胞。3.采用血管紧张素Ⅱ作为刺激因子,作用于大鼠肾小球系膜细胞,使其发生增殖。4.应用ELISA法测定10%FBS组及血管紧张素Ⅱ组纤维连接蛋白(FN)、Ⅳ型胶原的含量。5.采用逆转录-聚合酶链反应技术(RT-PCR)观察各组间MMP-3及TIMP-1的mRNA的表达情况。结果:1.血管紧张素Ⅱ组FN及Ⅳ型胶原的分泌量显著增加,同时与10%FBS组相比有显著差异(P<0.05)。表明用血管紧张素Ⅱ造肾小球系膜细胞增殖模型是可以信赖的。2.复方1号组对MMP3有显著的促进作用,与病理组比较有显著差异(P<0.05)。其余各组虽有促进MMP-3分泌的趋势,但无统计学意义。3.三组小复方对TIMP-1的影响无统计学意义。结论:1.复方1号组有显著的促进MMP-3基因表达的作用。2.复方1号组通过促进MMP-3的基因表达,从而发挥延缓和防治肾小球硬化的作用。3.复方2号组与复方3号组对MMP-3的基因表达有促进趋势。更多还原

【Abstract】 Purpose:Research the effect of three groups of chinese medicine compound prescriptions on expression of glomerular mesangial cells matrix metalloproteinases-3 (MMP-3) and tissue inhibitor of metallo proteinases-1 (TIMP-1), and probe into the effect mechanism of the effective medicine potency of delaying and preventing the glomerular sclerosis process.Material and method:1. Make three groups of chinese medicine small compound prescriptions and pharmacological blood serum of normal group by applying blood serum pharmacology method.2. Bring up rat glomerular mesangial cells by applying in vitro-culture method.3. Using AngiotensinⅡas a stimulating factor on rats mesangial cells, in order to make the cells to proliferate。4. Measure the content of fibronectin (FN) and collagen type IV in 10% FBS group and Angiotensin II group.5. Observe the expression situation of mRNA of MMP-3 and TIMP-1 among various groups by applying reverse transcription-polymerase chain reaction (RT-PCR).Results:1. Exudation of Angiotensin II group FN and collagen type IV increased significantly, and very different from that of 10% FBS group(P<0.05).2. Treatment group 1 has great promotion effect on MMP-3, and very different from that of pathology group(P<0.05), and although other groups have the current of promoting the exudation of MMP-3, it has no meaning to the statistics.3.The effect of three groups of small compound prescriptions on TIMP-1 has no meaning to the statistics. Conclusion:1. Treatment group 1 has the obvious effect of promoting the expression of MMP-3.2. Treatment group 1 will delay and prevent glomerular sclerosis by promoting the gene expression of MMP-3.3. Treatment group 2 and Treatment group 3 have the promotion effect on the gene expression of MMP-3.更多还原