肾血宁煎剂对原发性IgA肾病大鼠肾组织Ⅳ型胶原影响的实验研究

【作者】 李英南；

【导师】 马进；

【作者基本信息】 辽宁中医药大学 ， 中医内科学， 2008， 硕士

【摘要】 目的:通过肾血宁煎剂对原发性IgA肾病大鼠肾组织Ⅳ型胶原表达的影响,探讨其对原发性IgA肾病肾纤维化的影响及作用机理,寻求延缓或阻止IgA肾病病程发展的有效药物。方法:体重160±20g的健康雌性Wistar大鼠48只,实验前,均做尿蛋白、尿潜血检查,排除非健康实验动物对实验结果的影响。随机分为4组,即空白对照组、模型组、肾血宁煎剂组、代文(缬沙坦)组,每组12只。随机选取3组,建立IgA肾病模型。12周后,观察大鼠24h尿蛋白定量、尿红细胞计数、血清肌酐、血尿素氮、肾组织形态学和Ⅳ型胶原表达等指标。结果:1、模型组大鼠24h尿蛋白定量明显升高(P<0.05),肾血宁煎剂组、代文(缬沙坦)组大鼠24h尿蛋白定量增高的幅度显著低于模型组(P<0.05),肾血宁煎剂组与代文(缬沙坦)组相比,无显著性统计学意义(P>0.05)。2、模型组大鼠尿红细胞计数明显升高(P<0.05),肾血宁煎剂组大鼠尿红细胞计数增高的幅度显著低于模型组、代文(缬沙坦)组(P<0.05),代文(缬沙坦)组与模型组相比,无显著性统计学意义(P>0.05)。3、模型组大鼠血清肌酐、血尿素氮明显升高(P<0.05),肾血宁煎剂组、代文(缬沙坦)组大鼠血清肌酐、血尿素氮增高的幅度显著低于模型组(P<0.05),肾血宁煎剂组与代文(缬沙坦)组相比,无显著性统计学意义(P>0.05)。4、图像分析:模型组大鼠肾组织Ⅳ型胶原(Col一Ⅳ)平均光密度值明显升高(P<0.05),肾血宁煎剂组、代文(缬沙坦)组大鼠肾组织Ⅳ型胶原(Col一Ⅳ)平均光密度值增高的幅度显著低于模型组(P<0.05),肾血宁煎剂组与代文(缬沙坦)组相比,无显著性统计学意义(P>0.05)。5、病理:⑴HE染色结果:模型组大鼠肾小球明显肥大,肾小球系膜细胞增生伴系膜基质增多,部分肾小球节段性硬化,球囊壁粘连,肾小管上皮细胞空泡变性、肥大、增生,部分肾小管萎缩,肾间质有不同程度的单个核炎症细胞浸润和纤维化,肾血宁煎剂组、代文(缬沙坦)组大鼠较模型组程度明显减轻,且肾血宁煎剂组较代文(缬沙坦)组程度轻。⑵免疫荧光结果:模型组、肾血宁煎剂组、代文(缬沙坦)组大鼠肾小球基底膜和系膜区均有亮绿色阳性染色沉积,肾血宁煎剂组、代文(缬沙坦)组的亮绿色阳性染色沉积较模型组少,肾血宁煎剂组的亮绿色阳性染色沉积较代文(缬沙坦)组少。⑶免疫组化染色结果:空白对照组肾组织中Ⅳ型胶原(Col一Ⅳ)在肾小球毛细血管基底膜和系膜区、肾小管上皮细胞基底膜轻度表达,阳性染色为棕黄色颗粒,模型组中肾小球毛细血管基底膜和系膜区、肾小管上皮细胞基底膜及间质可见大量棕黄色颗粒沉积,肾血宁煎剂组、代文(缬沙坦)组棕黄色颗粒沉积较模型组明显减少,且肾血宁煎剂组较代文(缬沙坦)组程度轻。结论:1.肾血宁煎剂有降低原发性IgA肾病大鼠24小时尿蛋白、尿红细胞、血清肌酐、血尿素氮的作用。2.肾血宁煎剂有降低原发性IgA肾病大鼠Ⅳ型胶原表达的作用。3.肾血宁煎剂具有延缓IgA肾病病程发展的作用。其机制可能是通过降低Ⅳ型胶原表达,减轻ECM的过度蓄积,减轻了肾小球硬化和肾小管-间质纤维化,从而减轻尿蛋白、血尿,保护了肾功能。更多还原

【Abstract】 Goal:Through Shenxuening Jianji to primary IgA nephrosis big mouse’s kidney organization in typeⅣCollagen expression influence, discusses it to the primary IgA nephrosis kidney fibrosis influence and the action mechanism, seeks the effective medicine to postpone or prevent the IgA nephrosis course development.Method:Body weight 160±20g healthy female Wistar big mouse 48, before the experiment, makes the urine protein, the urine occult blood inspection, removes the non-health experimental animal to the experimental result influence. Divides into 4 groups stochastically, namely blank control group, model group, Shenxuening Jianji group, Diovan (Valsartan) group, each group of 12. Selects 3 groups stochastically, establishes the IgA nephrosis model. 12 weeks later, observes the indicators about the big mouse 24h urine protein quota, the urine red cell count, the serum creatinine, the blood urea nitrogen, Renal morphology and expression of typeⅣcollagen ,and so on.Finally:1、The model group big mouse 24h urine protein quota is significantly increased (P <0.05),the rate of increase of the Shenxuening Jianji group、the Diovan (valsartan) group 24h urine protein quota levels is significantly lower than the rate of the model group (P <0.05),Compared the Shenxuening Jianji group with the Diovan (valsartan) group, there is no statistically significant (P> 0.05).2、The model group big mouse urine red cell count is significantly increased (P <0.05),the rate of increase of the Shenxuening Jianji group urine red cell count levels is significantly lower than the rate of the model group、the Diovan (valsartan) group (P <0.05),Compared the Diovan (valsartan) group with the model group, there is no statistically significant (P> 0.05).3、The model group big mouse serum creatinine、blood urea nitrogen is significantly increased (P <0.05),the rate of increase of the Shenxuening Jianji group、the Diovan (valsartan) group serum creatinine, blood urea nitrogen levels is significantly lower than the rate of the model group (P <0.05),Compared the Shenxuening Jianji group with the Diovan (valsartan) group, there is no statistically significant (P> 0.05).4、Image Analysis:The model group big mouse kidney organization typeⅣCollagen(Col一Ⅳ)average optical density value is significantly increased (P <0.05),the rate of increase of the Shenxuening Jianji group、the Diovan (valsartan) group kidney organization typeⅣCollagen(Col一Ⅳ)average optical density value levels is significantly lower than the rate of the model group (P <0.05),Compared the Shenxuening Jianji group with the Diovan (valsartan) group, there is no statistically significant (P> 0.05).5、Pathology:⑴HE staining results: The model group big mouse glomerular is obviously hypertrophy, glomerular mesangial cells hyperplasia with Mesangial matrix increase, Part of glomerular Segmental sclerosis, ball wall adhesion, Renal tubular epithelial cells degeneration, hypertrophy, hyperplasia, Some tubular atrophy, Renal interstitial have varying degrees of mononuclear inflammatory cell infiltration and fibrosis, the Shenxuening Jianji group、the Diovan (valsartan) group compared to the model group is reduced significantly, and the Shenxuening Jianji group than the Diovan (valsartan) group level light.⑵Immunofluorescence Results: The model group, Shenxuening Jianji group, the Diovan (Valsartan) group big mouse glomerular basement membrane and mesangial area have bright green positive staining deposition, Shenxuening Jianji group, the Diovan (Valsartan) group bright green positive staining deposition less than the model group, and Shenxuening Jianji group bright green positive staining deposition less than the Diovan (Valsartan) group.⑶Immunohistochemical staining results: In the blank control group kidney organization ,typeⅣCollagen(Col一Ⅳ)has the Mild expression in the glomerular basement membrane and mesangial area, the basement membrane proximal tubular epithelial cells, positive staining for the brown granules ,and in the model group, glomerular basement membrane and mesangial area,renal tubular epithelial cells and the basement membrane mesenchymal show a large number of brown particle deposition. Shenxuening Jianji group, the Diovan (Valsartan) group brown granules deposition is significantly reduced compared to the model group,and Shenxuening Jianji group than the Diovan (Valsartan) group level light.Conclusion:1、The Shenxuening Jianji has the function of reducing the IgA nephrosis big mouse 24-hour urine protein and red blood cells, serum creatinine, blood urea nitrogen.2、The Shenxuening Jianji has the function of reducing the primary IgA nephrosis big mouse expression of typeⅣcollagen.3、The Shenxuening Jianji has the function of delaying the IgA nephrosis course development. Its mechanism may be by reducing expression of typeⅣcollagen,to reduce the excessive accumulation of ECM, reduce glomerulosclerosis and tubular - interstitial fibrosis, thereby reduce urine protein、hematuria, and Protect renal function.更多还原