蝉花菌丝延缓肾小球硬化的作用机制研究

The Mechanism Study of Cordyceps Sobolifera Mycelium Preventing the Progression of Glomerulosclerosis

目的:探讨蝉花菌丝延缓实验大鼠肾小球硬化作用的可能机制,观察和评价蝉花菌丝对实验大鼠慢性 肾衰竭的治疗效果。方法:对大鼠行5/6肾切除,建立肾小球硬化模型,并用蝉花菌丝对其治疗42d,其间观察大鼠的 尾动脉血压和测定24h尿蛋白定量,并在实验结束时检测各种生化指标(Scr、BUN、Alb、TG、TC、Hb、HCT、RBC)和取 材作病理分析,行HE染色,采用病理图像分析系统计算肾小球平均损伤面积的百分率;采用免疫组化和原位杂交技 术进行Col-Ⅳ、PAI-1、u-PA、u-PAmRNA和PAI-1mRNA免疫染色,应用计算机图像分析系统对其阳性表达 结果进行分析,并对模型对照组中Col-Ⅳ、PAI-1、u-PA蛋白表达作相关分析。结果:蝉花菌丝与模型组比较能明 显降低大鼠肾重/体重比值,24h尿蛋白量,降低大鼠BUN和Scr(P<0.01);模型组大鼠尾动脉血压在42d里均明显 高于蝉花菌丝组(P<0.05);蝉花菌丝组大鼠红细胞计数、血红蛋白、红细胞压积和血浆白蛋白均高于模型组大鼠 (P<0.05);免疫组化显示蝉花菌丝能减少大鼠肾小球中PAI-1蛋白的表达,提高u-PA的表达,与模型组比较有统 计学差异(P<0.01),而模型组大鼠肾小球内Col-Ⅳ的表达明显强于蝉花菌丝组(P<0.01);原位杂交技术显示 u-PAmRNA和PAI-1mRNA主要表达定位在肾小球系膜区、毛细血管襻?

Objective:To explore the mechanism of Cordyceps Sobolifera Mycelium (CSM) preventing the progression in the rat glomerulosclerosis model. To observe and evaluate the results of CSM used to treat the experimental rats with CRF.Methods:The glomerulosclerosis model was produced in rats by 5/6 renal mass ablation. Cordyceps Sobolifera Mycelium (CSM) was used to treat the glomerulosclerosis in forty-two days. The arteriotony in the rat-tail and 24 hours' urinary albumin quantity were detected in forty-two days. The serum biochemistry assay (Scr,BUN,Alb,TG,TCH,Hb,Hct,RBC) and renal tissue samples were obtained after rats were sacrificed. The sections were stained with hematoxylin and eosin. The percentage of injury area in the glomeruli was counted by the analytic system of pathological image. Immunohistochemistry and in situ hybridization were performed to detect the expression of PAI-1, u-PA,Col-Ⅳ,PAI-1 mRNA and u-PA mRNA in the glomeruli. The positive results were evaluated by a computer analytic system of pathological image. The relative analysis was made between control group and treated group for the expression of u-PA, PAI-1 and Col-Ⅳ protein in the glomeruli.Results:The renal weight/body weight, 24 hours' urinary albumin quantity and the blood urea nitrogen (BUN) and serum creatinine (Scr) of rats in CSM treated group were lower than those in the control group (P<0.01 respectively). The arteriotony in the rat-tail in the control group was higher than that in CSM treated group in the day 42 (P<0.05).The RBC,Hb,Hct and Alb in CSM treated group were higher than those in the control group (P<0.05 respectively).Immunohistochemical results showed that the expression of u-PA was upregulated and the expression of PAI-1 was downregulated in the CSM treated group compared with those in the control group (P<0.01).The expression of Col-Ⅳ in the control group was higher than that in the CSM treatment group (P<0.01). In situ hybridization results showed that the expression of u-PAmRNA and PAI-1mRNA localized in the mesangical area of glomeruli,capillary loops and part of visceral epithelial cells.The expression of u-PA mRNA in CSM treated group was higher than that in the control group (P< 0.01).But the expression of PAI-1 mRNA was lower than that in the control group (P<0.05).In the control group the expression of PAI-1 was closely related to that of Col-Ⅳ(r=0.932,P<0.01)Conclusion:CSM treatment could alleviate the hypertrophy of glomeruli and accumulation of Col-Ⅳ, and prevent the progression of CRF. The mechanism might be linked with its upregulation of u-PA mRNA expression and downregulation of PAI-1 mRNA expression.