›› 2012, Vol. 32 ›› Issue (1): 1-.doi: 10.3969/j.issn.1674-8115.2012.01.001

• 论著(基础研究) •    下一篇

冬虫夏草制剂拮抗5/6肾切除大鼠肾脏纤维化作用研究

郭山脉, 仲 芳, 周 桥, 卢 颖, 郝 旭, 王伟铭, 陈 楠   

  1. 上海交通大学 医学院附属瑞金医院肾脏科, 上海 200025
  • 出版日期:2012-01-28 发布日期:2012-01-29
  • 通讯作者: 王伟铭, 电子信箱: wweiming@medmail.com.cn。
  • 作者简介:郭山脉(1983—), 女, 硕士生;电子信箱: gshanmai@126.com。
  • 基金资助:

    国家自然科学基金(30270613,30771000);上海市重点学科项目(T0201);上海市科委重点项目(07JC14037);上海市科委科研计划项目(09dz1900502);2011年上海交通大学医学院“博士创新基金”(BXJ201107)

Renal protective effect of Cordyceps sinensis on 5/6 nephrectomy-induced renal fibrosis in rats

GUO Shan-mai, ZHONG Fang, ZHOU Qiao, LU Ying, HAO Xu, WANG Wei-ming, CHEN Nan   

  1. Department of Nephrology, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200025, China
  • Online:2012-01-28 Published:2012-01-29
  • Supported by:

    National Natural Science Foundation of China, 30270613, 30771000;Shanghai Leading Academic Discipline, T0201;Shanghai Science and Technology Committee Foundation, 07JC14037, 09dz1900502;Shanghai Jiaotong University School of Medicine Foundation, BXJ201107

摘要:

目的 构建5/6肾切除大鼠慢性肾脏病模型,研究冬虫夏草制剂对肾脏纤维化的拮抗作用。方法 48只雄性SD大鼠随机分为假手术组(SO组)、5/6肾切除组(OP组)和5/6肾切除术后冬虫夏草制剂治疗组(CS组),每组16只。分别于肾切除术后4、8周,采集血、尿样本,处死大鼠后留取肾组织标本,测定各组大鼠血清肌酐(SCr)、尿素氮(BUN)、白蛋白(Alb)和尿白蛋白肌酐比(UACR),观察肾脏组织病理改变和超微结构变化,免疫组织化学法检测各组大鼠肾脏组织中细胞外基质纤维连接蛋白(FN)和Ⅲ型胶原(ColⅢ)以及成纤维细胞特异性蛋白1(FSP-1)和α-平滑肌动蛋白 (α-SMA)的表达,采用RT-PCR技术检测肾脏组织中FN、Col-Ⅲ和α-SMA 的mRNA表达。结果 与SO组比较,OP组肾脏组织病变明显,表现为炎症细胞浸润,细胞外基质增多,足突融合及线粒体肿胀、空泡变性甚至嵴溶解、消失,自噬小体形成,且术后8周较术后4周的病变更为严重;与SO组比较,OP组SCr、BUN和UACR均显著升高,Alb则明显降低,肾脏组织中FSP-1、FN、Col-Ⅲ、α-SMA的蛋白表达及FN、Col-Ⅲ和α-SMA 的mRNA表达均显著增加,上述改变术后8周较术后4周更为明显。术后8周,与OP组比较,CS组肾脏组织病理变化明显减轻,线粒体损伤明显缓解,UACR明显降低,而Alb升高显著,肾脏组织中FSP-1、FN、Col-Ⅲ、α-SMA的蛋白表达及FN、Col-Ⅲ和α-SMA的mRNA表达明显减少。结论 冬虫夏草制剂可通过减轻线粒体损伤和细胞外基质沉积等机制,拮抗5/6肾切除大鼠的肾脏纤维化程度。

关键词: 冬虫夏草制剂, 肾脏纤维化, 细胞外基质, 线粒体损伤

Abstract:

Objective To establish the rat model of 5/6 nephrectomy-induced chronic renal disease, and investigate the protective effect of Cordyceps sinensis on renal fibrosis. Methods Forty-eight male SD rats were randomly divided into sham-operation group (SO group), 5/6 nephrectomy group (OP group) and5/6 nephrectomy + Cordyceps sinensis treatment group (CS group), with 16 rats in each group. Blood and urine samples were collected 4 weeks and 8 weeks after nephrectomy, and rats were sacrificed for renal tissue samples. Serum creatinine (SCr), blood urea nitrogen (BUN), albumin (Alb) and urinary albumin to creatinine ratio (UACR) were measured, pathological changes and ultrastructure changes of renal tissues were observed, the expression of extracellular matrix fibronectin (FN), collagen type Ⅲ (Col-Ⅲ), fibroblast-specific protein 1 (FSP-1) and α-smooth muscle actin (α-SMA) in renal tissues was detected by immunohistochemistry, and the expression of FN, Col-Ⅲ and α-SMA mRNA in renal tissues was determined by RT-PCR. Results Compared with SO group, significant pathological changes were observed in OP group 4 weeks after surgery, which were characterized by infiltration of inflammatory cells, more deposition of extracellular matrix, podocyte foot process effacement, mitochondrial swelling, vacuolization, distorted cristae and autophagosome formation, and these changes were more significant 8 weeks after surgery. Compared with SO group, SCr, BUN and UACR were significantly higher, Alb was significantly lower, the expression of FSP-1, FN, Col-Ⅲ and α-SMA protein and FN, Col-Ⅲ and α-SMA mRNA in renal tissues was significantly higher in OP group 4 weeks after surgery, and these differences were more significant 8 weeks after surgery. Compared with OP group, the pathological changes of renal tissues obviously lessened, mitochondrial damage significantly alleviated, UACR significantly decreased, Alb significantly increased, and the expression of FSP-1, FN, Col-Ⅲ and α-SMA protein and FN, Col-Ⅲ and α-SMA  mRNA in renal tissues significantly decreased in CS group 8 weeks after surgery. Conclusion Cordyceps sinensis ameliorates 5/6 nephrectomy-induced renal fibrosis of rats by relieving mitochondrial damage and reducing deposition of ECM.

Key words: Cordyceps sinensis, renal fibrosis, extracellular matrix, mitochondrial damage