通脉口服液对实验性DN模型大鼠肾组织Col-Ⅳ及MMP-9/TIMP-1的影响

Effects of Tongmai Oral Solution on Expression of Col-Ⅳ and MMP-9/TIMP-1 in Kidney of DN Rats

目的:观察通脉口服液对实验性DN模型大鼠肾组织Col-Ⅳ蛋白表达及MMP-9/TIMP-1作用的影响。方法:SD雄性大鼠,予STZ单次腹腔注射及高脂饲料造模,筛选DN成模大鼠,随机分组,药物干预;6周后处死大鼠取肾组织,以免疫组化方法测定Col-Ⅳ、MMP-9及TIMP-1的蛋白表达,计算积分光密度及面密度比值。结果:Col-Ⅳ表达主要定位于肾小球;MMP-9表达主要定位于肾小球,肾间质内有少量表达;TIMP-1表达主要定位于肾小球,肾小管及间质区内有少量表达。通脉组Col-Ⅳ免疫组化染色面密度及积分光密度较模型组降低,MMP-9免疫组化染色面密度及积分光密度较模型组升高,差异有统计学意义(P<0.01或P<0.05);MMP-9/TIMP-1比值与Col-Ⅳ免疫组化染色面积存在负相关(r=-0.919,P<0.01)。结论:益气活血中药复方通脉口服液可能通过调节肾组织MMP-9/TIMP-1蛋白表达,改善Col-Ⅳ代谢,发挥DN肾脏保护作用。

Objective：To observe the effects of the TongMai oral solution on the expression of Col-Ⅳ and MMP-9/TIMP-1 in renal tissue of DN rats.Methods：SD male adult rats of specified-pathogen free were injected with streptozotocin （STZ） intraperitoneally and given high-fat food on alternate days to establish animal model of Diabetes Mellitus.After the model was set up,the model rats were randomly divided into model group and medicated group treated by Tongmai oral solution.After 6 weeks,immunohistochemical staining method was used for detecting the expressions of type Ⅳ collagen,MMP-9,TIMP-1 in kidneys of rat model.Another group was set for experimental control,consisted of normal SD rats.Results：The type Ⅳ collagen was located in the glomerulus.Strong positive expression was found in model group.According to the outcomes of immunohistology,the values of area density and photodensity of positively stained area were less than those of model group with statistical significance（P〈0.01）.The expression of MMP-9 was located mainly in glomerulus and sparse expression could be seen in renal interstitium.It＇s more weakly expressed in model group than in normal control group.The values of area density and photodensity in MMP-9 positively stained area were higher than those of model group（P〈0.01 or P〈0.05）.Expression of TIMP-1 was mainly located in glomerulus with weakly expression located in renal tubule and interstitium.The ratio of MMP-9/TIMP-1 and photodensity of type Ⅳ collagen changed in reverse direction（r=-0.919,P〈0.01）.Conclusion：The mechanism that Tongmai Oral Solution ameliorated the structure and function damage of DN model rats may be due to regulation of the expression of MMP-9/TIMP-1 and increase of degeneration of type Ⅳ collagen.