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  • 孙亮亮,李文桐,冯晓云,张兰予,邹俊杰,石勇铨,刘志民.通络方剂减轻糖尿病大鼠肾小球足细胞损伤[J].第二军医大学学报,2012,33(2):165-169    [点击复制]
  • SUN Liang-liang,LI Wen-tong,FENG Xiao-yun,ZHANG Lan-yu,ZOU Jun-jie,SHI Yong-quan,LIU Zhi-min.Protective effects of Tongluo recipe against injury of glomerular visceral epithelial cells in diabetic rats[J].Acad J Sec Mil Med Univ,2012,33(2):165-169   [点击复制]
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通络方剂减轻糖尿病大鼠肾小球足细胞损伤
孙亮亮1,李文桐2,冯晓云1,张兰予1,邹俊杰1,石勇铨1,刘志民1*
0
(1.第二军医大学长征医院内分泌科,上海 200003
2.上海警备区第一干休所,上海 200040
*通信作者)
摘要:
目的 观察通络方剂(TLR)和抗氧化剂α-硫辛酸减轻糖尿病大鼠肾小球足细胞损伤的作用,探讨糖尿病肾病足细胞损伤的可能机制。方法 通过高脂饮食和腹腔注射链脲佐菌素(35 mg/kg)制作糖尿病大鼠模型,将成模大鼠随机分为糖尿病组(DM组,8只)、糖尿病α-硫辛酸干预组(DM+ALA组,8只)、糖尿病TLR干预组(DM+TLR组,8只),另设正常对照组(CON组,8只)。DM+ALA组大鼠予以α-硫辛酸(0.1 g·kg-1·d-1)灌胃,DM+TLR组大鼠予以TLR (0.4 g·kg-1·d-1)灌胃,DM组和CON组每日均在同一时间予同体积蒸馏水一次性灌胃。干预12周后测定各组大鼠肾质量/体质量比值及24 h尿蛋白定量、BUN、Cr、Alb/Cr等肾功能指标;采用体视学方法 计数足细胞和计算足细胞相对密度;采用蛋白质印迹分析法检测足细胞nephrin蛋白的表达;测定血浆和肾皮质中超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性和脂质过氧化产物丙二醛(MDA)含量。结果 12周时,与CON组相比,DM组大鼠肾质量/体质量增大,24 h尿蛋白、尿白蛋白/肌酐升高(P均<0.01);足细胞计数减少、足细胞密度降低且nephrin表达减少(P均<0.01);DM+ALA组和DM+TLR组大鼠肾质量/体质量、24 h尿蛋白、尿白蛋白/肌酐均低于DM组(P<0.01),足细胞计数、足细胞相对密度和nephrin表达量均高于DM组(P<0.01,P<0.05)。与CON组相比,DM组、DM+ALA组和DM+TLR组大鼠在观察终点血浆和肾皮质SOD、GSH-Px活性均降低(P<0.01,P<0.05),MDA含量则升高(P<0.01,P<0.05);与DM组相比,DM+ALA组和DM+TLR组血浆和肾皮质SOD、GSH-Px活性增加(P均<0.01),MDA含量降低(P<0.01,P<0.05)。DM+ALA组与DM+TLR组之间差异无统计学意义。 结论 TLR可改善糖尿病大鼠肾小球足细胞损伤,可能与其抗氧化应激作用有关。
关键词:  通络方剂  糖尿病肾病  足细胞  氧化性应激
DOI:10.3724/SP.J.1008.2012.0165
投稿时间:2011-11-03修订日期:2012-01-04
基金项目:国家重点基础研究发展计划(“973”计划,2005CB523304).
Protective effects of Tongluo recipe against injury of glomerular visceral epithelial cells in diabetic rats
SUN Liang-liang1,LI Wen-tong2,FENG Xiao-yun1,ZHANG Lan-yu1,ZOU Jun-jie1,SHI Yong-quan1,LIU Zhi-min1*
(1. Department of Endocrinology, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China
2. The First Sanatorium for Retired Cadre, Garrison Command of Shanghai, Shanghai 200040, China
*Corresponding author.)
Abstract:
Objective To observe the protective effect of α-lipoic acid (ALA) and Tongluo recipe(TLR) against injury of glomerular visceral epithelial cells in diabetic rats, so as to discuss the possible mechanism for injury of glomerular visceral epithelial cells in diabetes. MethodsRat diabetes model was induced by high-fat diet and a single intraperitoneal injection of streptozotocin (35 mg/kg). The diabetic model rats were randomly divided into three groups (n=8): diabetes mellitus (DM) group,DM+TLR and DM+ALA group; another 8 rats served as normal controls (CON group). Rats in the DM+TLR group received 0.4 g·kg-1·d-1 TLR and DM+ALA group received 0.1 g·kg-1·d-1 ALA throughout the experiment. Rats in the DM group and control (CON) group were given celiac perfusion of distilled water once at the same time and volume. After 12 weeks of treatment, kidney mass/body mass index, 24 h urinary protein, blood urea nitrogen, and urinary albumin/Cr were determined in each group. The number and the density of podocytes in glomerulis were measured by stereological dissector/fractionator methods. The expression of nephrin in renal cortical tissue was detected by Western blotting analysis. The activity of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and the content of malondialdehyde (MDA)were measured in the plasma and renal cortical tissues. ResultsAfter 12 weeks, the kidney mass/body mass index, 24 h urinary protein and urinary albumin/Cr in DM group were significantly increased compared to control group (P<0.01); the number and density of podocytes in the glomerulis and nephrin expression in DM group were significantly decreased (P<0.01). The kidney mass/body mass index, 24 h urinary protein and urinary albumin/Cr in DM+TLR and DM+ALA groups were significantly decreased compared to DM group (P<0.01); the number and density of podocytes and nephrin expression in DM+TLR and DM+ALA groups were significantly increased compared to DM group (P<0.01, P<0.05). GSH-Px and SOD activities were significantly decreased in the plasma (P<0.01) and renal cortical tissue (P<0.01) in DM, DM+ALA and DM+TLR groups compared to those in the CON group, and the content of MDA was significantly increased (P<0.01, P<0.05). GSH-Px and SOD activities were significantly increased in the plasma and renal cortical tissue (P both<0.01) and MDA content was significantly decreased in DM+TLR and DM+ALA groups (P<0.01, P<0.05)compared to those in the DM group. No significant differences were found between DM+ALA group and DM+TLR group concerning the above parameters. ConclusionTLR can ameliorate podocyte injury in diabetic rats, which might be associated with its anti-oxidative effect.
Key words:  Tongluo recipe  diabetic nephropathies  podocytes  oxidative stress