芡实对糖尿病肾病大鼠肾组织MMP-9、TIMP-1及Collagen Ⅳ表达的影响

Effect of Euryale Ferox on the Expression of MMP-9,TIMP-1 and Collagen Ⅳ in the Renal Tissues of Diabetic Nephropathy Rats

目的:探讨芡实对糖尿病肾病大鼠肾组织MMP-9、TIMP-1及CollagenⅣ表达的影响。方法:健康雄性Wistar大鼠65只,随机选取10只为正常对照组(N组);其余一次性腹腔注射链脲佐菌素(STZ)制作DN模型,造模成功后,随机分为DN组、芡实低、中、高剂量组及氯沙坦钾组,均采用灌胃给药,N组和DN组给予等体积蒸馏水。12周后检测大鼠生化指标;HE、Masson、PAS染色观察肾脏病理变化;免疫组化法测定MMP-9、TIMP-1及CollagenⅣ在肾组织表达情况;采用实时荧光定量PCR技术检测肾组织中MMP-9、TIMP-1及CollagenⅣ基因的表达。结果:实验12周末,与N组比较,DN组大鼠肾小球肥大,细胞外基质积聚,血糖、24 h尿蛋白定量、血尿素氮(BUN)、肌酐(Scr)、尿白蛋白排泄率(UAER)、尿蛋白/尿肌酐(Up/Ucr)比值显著升高(P<0.05),肾组织MMP-9蛋白和mRNA表达降低(P<0.05),TIMP-1、CollagenⅣ蛋白和mRNA表达升高(P<0.05);与DN组比较,EM组、EH组及LP组大鼠病理改变减轻,24 h尿蛋白定量、Scr、BUN、UAER、Up/Ucr显著降低(P<0.05),肾组织MMP-9蛋白和mRNA表达显著升高(P<0.05),TIMP-1、CollagenⅣ蛋白和mRNA表达显著减少(P<0.05)。结论:芡实可能通过调节DN大鼠肾脏MMP-9、TIMP-1的表达,减少细胞外基质积聚,从而减轻肾小球硬化,减少蛋白尿,保护肾功能,延缓DN病变的进展。

Objective ：To investigate the effect of Euryale ferox on the expression of MMP - 9, TIMP - 1 and Collagen Ⅳ in renal tissues of diabetic ne -phropathy rats. Methods：65 Wistar male rats were used. 10 rats were selected randomly as the normal control group （N）, the rest of 55 rats were made DN models by injected streptozocin into abdominal cavity （STZ）, then these rats were randomly divided into 5 groups： diabetic nephropathic model group （DN）, 1- ow- dose Euryale ferox treatment group （EL）, medium - dose Euryale ferox treatm - ent group （ EM）, high - dose Euryale ferox treatment group （EH） and losartan pot - assium group （LP）, and 11 rats in every group, while rats were treated respecti -vely by intragastric administration for 12 weeks with the different dose of Eur - yale ferox and losartan potassium. Meanwhile rats of group N and group DN r - eceived the same volume dis- tilled water by garage. 24 h urinary protein, serum BUN, Scr, Urinary albumin excretion ratio （UAER） and the urinary protein to creatinine ratio （Up/Ucr） were measured after the research. The renal pathologica - 1 changes were observed by hematoxylin - eosin, Masson and Periodic acid Schif-f （PAS） staining. The expression of MMP-9, TIMP- 1 and Collagen Ⅳ were det -ected by im- munohistochemistry. The gene expressions of MMP - 9, TIMP - 1 and Collagen Ⅳ were measured by Real - time PCR. Results： Compared with group N, the renal glomerular became hypertrophic and the extracellular matrix accum - ulated in group DN. 24 h uri- nary protein, BUN, Scr, UAER and Up/Ucr also in - creased significantly in group DN （ P 〈 0.05 ） ; DN group has a decrease of mR - NA of MMP - 9 but an increase of the TIMP - 1 and Collagen Ⅳ expressions （ P 〈 0.05 ）. While the pathology changes were re- lieved, 24 h urinary protein, BU -N, Scr, UAER and Up/Ucr as well as the mRNA of MMP -9 increased but the decrease of the TIMP - 1 and Collagen Ⅳ expressions in group EM, EH and L - P （ P 〈 0.05 ）. Conclusion： Euryale ferox may be affected by the expression of MMP - 9 and TIMP - 1 in the DN rats kidney, to reduce the extracellular matrix accumulation, thus it may relieve glo- merular sclerosis and decrease proteinuria, meanwhile protect kidney function and delay the progress of diabetic nephropath- y.