液体培养蝉花菌丝对5/6肾切除大鼠肾小球硬化的干预作用

目的探讨人工液体培养法培育的蝉花菌丝对5/6肾切除大鼠肾小球硬化的干预作用及其作用机制。方法建立5/6肾切除SD大鼠肾小球硬化模型,随机分为模型对照组（模型组）、科素亚治疗组（科素亚组）、天然蝉花菌丝治疗组（天然组）、大剂量液体培养蝉花菌丝治疗组（液大组）和小剂量蝉花菌丝治疗组（液小组）,分别给予相应的干预措施干预42天;检测各组大鼠的血清肌酐（Scr）、尿素氮（BUN）、白蛋白（ALB）和24h尿蛋白定量;对大鼠残余肾做病理组织学分析,行HE染色、PAS染色,观察大鼠残肾组织形态学改变情况,计算肾小球硬化指数（GSI）,采用病理图像分析系统计算肾小球病理损害面积;采用免疫组化技术分别检测残余肾组织中IV型胶原（Col-IV）、纤维连接蛋白（FN）、转化生长因子-β1（TGF-β1）、基质金属蛋白酶-2（MMP-2）、基质金属蛋白酶抑制因子-2（TIMP-2）的表达,应用图像分析系统对其阳性表达结果进行定量分析。结果①模型组Scr、BUN、ALB及24h尿蛋白定量水平与假手术组比较,差异均有统计学意义（P〈0.05）;各给药组大鼠血Scr、BUN、ALB与模型组比较,差异均有统计学意义（P〈0.05）;给药组组间差异无统计学意义（P〉0.05）;科素亚组、天然组24h尿蛋白定量较模型组明显降低（P〈0.05）;液大组、液小组与模型组之间差异无统计学意义（P〉0.05）,给药组组间差异无统计学意义（P〉0.05）。②模型组与假手术组GSI及基质阳性面积比值比较,差异均有统计学意义（P〈0.05）;各给药组与模型组GSI及基质阳性面积比值比较,差异均有统计学意义（P〈0.05）;科素亚组与天然组、液大组GSI及基质阳性面积比值差异无统计学意义（P〉0.05）,与液小组差异有统计学意义（P〈0.05）;天然组与液大组GSI及基质阳性面积比值差异无统计学意义（P〉0.05）,与液小组差异有统计学意义（P〈0.05）;液大组与液小组基质阳性面积比值差异有统计学意义（P〈0.05）。③模型组与假手术组免疫组化染色阳性面积比值比较,差异均有统计学意义（P〈0.05）;各给药组与模型组比较,差异均有统计学意义（P〈0.05）;科素亚组与天然组结果相近,除MMP-2、TIMP-2染色阳性面积比值外,其他指标与液大组差异无统计学意义（P〉0.05）,与液小组各指标差异均有统计学意义（P〈0.05）;天然组与液大组、液大组与液小组各指标差异均有统计学意义（P〈0.05）。结论液体培养蝉花菌丝能改善慢性肾衰竭大鼠的肾功能,减轻肾组织内炎性细胞浸润和细胞增生,延缓肾小球硬化的进程。

Intervention of glomerulosclerosis following 5/6 nephrectomy by artificial cultured cordyceps sobolifera mycelium in rats

Objective To explore the mechanisms of artificial cultured cordyceps sobolifera mycelium（ACCSM） in preventing the progress of glomerulosclerosis following 5/6 nephrectomy in rats.Methods The glomerulosclerosis model was produced in rats by 5/6 nephrectomy.ACCSM was used to treat the glomerulosclerosis for forty-two days.The serum biochemistry assay（Scr,BUN,ALB） and 24-hours＇ urinary albumin were detected.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 pathologyical image,and glomerulosclerosis index（GSI） was counted too.Immunohistochemistry was performed to detect the expressions of Col-IV,FN,TGF-β1,MMP-2 and TIMP-2.The positive results were evaluated by using quantitative analysis method with the analytic system of pathological image.Results The serum creatinine（Scr） and blood urea nitrogen（BUN） of rats from the CSM and ACCSM treatment groups were lower than those of rats from the control group（P〈0.05）;the ALB in the CSM and ACCSM treatment groups were higher than those in the control group（P〈0.05）.Sections stained with HE and PAS and immunohistochemical method and image analysis showed that the expressions of Col-IV,FN,TGF-β1 and TIMP-2 in the ACCSM treatment group were downregulated and the expression of MMP-2 was upregulated（P〈0.05）.Conclusion ACCSM can prevent the progression of glomerulosclerosis in rats with 5/6 nephrectomy,relieve the expression of glomerulosclorosis and prevent the progression of chronic renal failure.