【作者】 张新；

【导师】 吕永曼；

【作者基本信息】 华中科技大学 ， 内科学， 2008， 硕士

【摘要】 目的本实验构建阿霉素肾硬化模型,通过曲尼司特给药干预,观察曲尼司特对实验动物肾组织形态学变化以及细胞外基质金属蛋白酶诱导因子(extracellular matrix metalloproteinase inducer ,CD147)和基质金属蛋白酶2(matrix metalloproteinase 2,MMP-2)表达的影响,初步探讨曲尼司特对阿霉素肾硬化大鼠肾脏病理的影响及作用机理,为临床防治肾小球纤维化提供理论依据。方法30只清洁级SD雄性大鼠,随机分为假手术组(S组)和手术组,手术组采用单侧左肾切除并阿霉素尾静脉注射方法建立大鼠肾小球硬化模型,造模成功后手术组大鼠随机分为模型组(M组)和曲尼司特治疗组(T组)。治疗组给予曲尼司特(400mg/kg/day),观察干预前后尿蛋白、血尿素氮(BUN)、肌酐(SCr)的变化,给药后8周处死所有大鼠,取右肾组织进行病理染色,在光镜下观察肾组织形态学变化;RT-PCR方法检测肾组织CD147和MMP-2mRNA的表达,并用免疫组化法观察肾组织CD147和MMP-2蛋白的表达,并进行图像分析。结果(1)肾组织PAS染色显示:S组未见明显病理改变;M组见多数肾小球表现为系膜细胞轻度弥漫性增生,伴节段性加重,细胞外基质显著增多,局灶节段性硬化,少部分可见球性硬化,毛细血管腔部分节段性塌陷,肾重/体重明显增高(P<0.05),肾小球硬化指数明显增高(P<0.05);与M组相比,T组病理改变明显减轻,肾小球系膜细胞和基质轻度节段性增生,毛细血管腔轻度狭窄,偶见局灶节段硬化,肾重/体重较模型组明显降低(P<0.05),肾小球硬化指数明显降低(P<0.05)。(2)免疫组化结果显示:M组CD147和MMP-2蛋白表达降低,与S组相比有统计学意义(P<0.05); T组上述表达增加,与M组相比有统计学意义(P<0.05)。(3)RT-PCR结果显示:M组CD147和MMP-2 mRNA表达降低,与S组相比有统计学意义(P<0.05);T组上述表达增加,与M组相比有统计学意义(P<0.05)。结论(1)在阿霉素肾硬化大鼠模型,曲尼司特能够减轻肾组织的病理损害,延缓肾小球硬化的进程;(2)曲尼司特能抑制CD147和MMP2 mRNA及其蛋白质的表达;(3)曲尼司特对肾小球硬化的防治作用可能与其提高CD147的表达从而提高MMP-2的表达有关。更多还原

【Abstract】 Objective In this experiment, we construct a rat model of Adriamycin-induced glomerulosclerosis and observe the effect of tranilast on the pathology change of kidney and the expression of CD147 and MMP-2 . And also we discusse the mechanism of Adriamycin-induced glomerulosclerosis in rats and provide some evidence for its prospective use in treatment of glomerular fibrosis.Methods Thirty Sprague-Dawley male rats were randomly divided into sham operation group(S group) and operation group. The operation group rats underwent unilateral nephrectomy of left kidney and intravenous injection of doxorubicin. Then the operation group rats were randomly divided into two groups: model group(M group), tranilast-treated group(T group). T group rats were fed tranilast(400mg/kg/day) and then albuminuria, creatinine, urea nitrogen, were detected. The rats were killed eight weeks later and the right renal tissues were examined by microscope. And the expression of CD147 and MMP-2 was studied by PT-PCR and immunohistochemistry. The intensity was analyaed by imagine quantitative analysis technique.Results (1)PAS staining of the renal tissue revealed that S group rats had no obvious pathological changes. M group rats tissue mainly revealed that mesangial cells mild to moderate segmental hyperplasia, extracellular matrix significantly increased, focal segmental sclerosis, and that a small portion of the ball sclerosis, capillary cavity collapse of the segmental renal weight / body weight significantly increased(P<0.05), glomerulosclerosis index increased significantly(P<0.05). Compared with M group, T group pathological changes reduced significantly, a few glomerular mesangial cells and stromal mild to moderate segmental increased capillary cavity see only a few mild stenosis, and occasionally focal segmental sclerosis, kidney weight / body weight compared model group decreased(P<0.05), glomerulosclerosis index decreased significantly(P<0.05). (2) Immunohistochemistry showed that group M CD147 and MMP-2 expression decreased, compared with the S group statistically significant(P<0.05), obove expression increased in the T group(P<0.05). (3) RT-PCR showed that in the M group CD147 and MMP-2 mRNA expression, compared with the S group statistically significant(P<0.05), obove expression increased in the T group(P<0.05).Conclusions (1) In the doxorubicin rat model of renal sclerosis, tranilast can reduce the renal pathological damage and delay the process of glomerulosclerosis. (2)Tranilast can inhibit the expression of CD147 and MMP-2 mRNA and protein. (3) Effects of tranilast on glomerular sclerosis may be related to the preventive effect of increased expression of CD147 to enhance the expression of MMP-2.更多还原