固体培养蝉花菌丝延缓肾小球硬化作用及其机制的实验研究

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

目的 :探讨人工固体培养法培育的蝉花菌丝延缓实验大鼠肾小球硬化作用的机制 ,观察和评价其对肾小球硬化的疗效。方法 :对大鼠行 5 / 6肾切除 ,建立肾小球硬化动物模型 ,采用天然、不同剂量固体培养蝉花菌丝治疗4 2d ,观察各组大鼠的血肌酐、尿素氮、白蛋白和 2 4h尿蛋白定量 ;对大鼠肾脏做病理组织学分析 ,行HE染色、PAS染色 ,采用病理图像分析系统计算肾小球病理损害面积的百分率 ;采用免疫组化技术进行Col -Ⅳ、FN、LN、MMP - 2、TIMP - 2免疫染色 ,应用图像分析系统对其阳性表达结果进行定量分析。结果 :与模型组比较 ,天然、固体培养蝉花菌丝能明显减少大鼠血肌酐、尿素氮上升程度 (P <0 .0 5 ) ,天然、大剂量固体培养蝉花菌丝组血白蛋白显著升高 (P <0 .0 5 ) ,2 4h尿蛋白量不同程度减少 ;HE染色、PAS染色肾组织病理片检查和免疫组化染色显示 ,固体培养蝉花菌丝能明显减轻大鼠肾小球硬化程度 ,减少肾小球内Col-Ⅳ、FN、LN、TIMP - 2的表达 ,提高MMP - 2的表达 ,与模型组比较有统计学差异 (P <0 .0 5 )。结论 :固体培养蝉花菌丝能延缓大鼠肾小球硬化进程和慢性肾衰竭进展速度。

Objective:To explore the mechanism of Artificial Culture Cordyceps Sobolifera Mycelium (ACCSM) preventing the progression in the rat glomerulosclerosis model.To observe and evaluate the results of CSM used to cure the experimental rats with CRF.Methods:The glomerulosclerosis model was produced in rats by 5/6 renal mass ablation. ACCSM was used to cure the glomerulosclerosis in forty-two days. The serum biochemistry assay (Scr,BUN,Alb) and 24 hours' urinary albumin quantity were detected in forty-two days. Renal samples were obtained after rats received ACCSM. Sections were stained with HE and PAS. The percentage of injury area in the glomeruli of kidney was counted by the analytic system of pathology image. Immunohistochemistry was performed to detect the expression of Col-Ⅳ,FN,LN,MMP-2,TIMP-2.The positive results were evaluated by using quantitative analysis method with the analytic system of pathology image.Results:The serum creatinine (Scr) and blood usea nitrogen (BUN) of rats from the CSM and ACCSM treatment group were lower than those of rats from the control group (P<0.05). The ALB in the CSM and big dosage ACCSM treatment group were higher than those in the control group (P<0.05), and 24 hours' urinary albumin quantity was decreased.Sections stained with HE and PAS and immunohistochemical method and image analysis showed that the expression of Col-Ⅳ,FN,LN,TIMP-2 in the ACCSM treatment group was downregulated and the expression of MMP-2 was upregulated compared with the control group (P<0.05).Conclusion:ACCSM could relieve the expression of glomerulosclerosis and prevent the progression of chronic renal failure (CRF).