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通络方剂对糖尿病大鼠肾脏氧化应激的影响
张春阳,邹俊杰,石勇铨,曲卫,孙亮亮,刘志民*
0
(第二军医大学长征医院内分泌科,上海 200003)
摘要:
目的:研究两种剂型通络方剂(超微粉和微粉)对糖尿病SD大鼠肾脏氧化应激的影响。方法:链脲佐菌素(60 mg/kg)腹腔注射诱导糖尿病大鼠。1周后测血糖≥16.7 mmol/L为糖尿病,将大鼠随机分为糖尿病对照组(DMC,n=7)、通络方剂超微粉组(DM+UT,n=8)和通络方剂微粉组(DM+FT,n=8)3组,同时取8只大鼠作为正常对照组(NC)。DM+UT组和DM+FT组按每天1 g/kg UT或FT灌胃,NC和DMC组给予等量的生理盐水溶液。干预4周后,测肾质量/体质量(KW/BW)和24 h尿蛋白(24 h UP),比色法检测肾皮质抗氧化酶和丙二醛(MDA),实时PCR法检测NADPH氧化酶亚基p47phox和p22phox mRNA表达的改变。结果:与正常大鼠相比,糖尿病大鼠肾皮质MDA含量、24 h UP和KW/BW升高,总超氧化物歧化酶(TSOD)活性降低,谷胱甘肽过氧化物酶(GSH-Px)活性升高,均有统计学意义;正常大鼠与糖尿病大鼠间过氧化氢酶(CAT)活性差异无统计学意义。与DMC组相比,两个通络方剂干预组糖尿病大鼠肾皮质MDA、24 h UP和KW/BW明显降低;TSOD和GSH-Px活性增强,差异均有统计学意义。DMC组和两个通络方剂组间CAT活性差异无统计学意义。其中DM+UT组对24 h UP、KW/BW、MDA、TSOD和GSH-Px的作用较DM+FT组更明显,差异有统计学意义。与正常大鼠相比,糖尿病大鼠肾皮质NADPH氧化酶亚基p47phox mRNA表达增强,差异有统计学意义。与DMC组相比,两个通络方剂干预组p47phox mRNA表达降低,差异有统计学意义,但两干预组间差异无统计学意义。在各实验组间NADPH氧化酶亚基p22phox mRNA表达的差异无统计学意义。结论:通络方剂能增强糖尿病大鼠肾脏TSOD和GSH-Px活性,降低NADPH氧化酶p47phox mRNA的表达,可能因此抑制肾脏氧化应激水平,减轻糖尿病大鼠早期的肾脏损伤。
关键词:  通络方剂  糖尿病    抗氧化酶  NADPH氧化酶
DOI:10.3724/SP.J.1008.2008.00940
投稿时间:2007-11-13修订日期:2008-01-30
基金项目:国家重点基础研究规划(“973”计划)(2005CB523304).
Effect of Tongluo recipe on oxidative stress in kidneys of diabetic rats
ZHANG Chun-yang, ZOU Jun-jie, SHI Yong-quan, QU Wei, SUN Liang-liang,LIU Zhi-min*
(Department of Endocrinology, Changzheng Hospital, Second Military Medical University, Shanghai 200003,China)
Abstract:
Objective:To study the effect of two types of Tongluo recipe (ultrafine type and fine type) on the level of oxidative stress in kidneys of diabetic Sprague-Dawley rats. Methods: Experimental diabetes was induced by intraperitoneal injection of streptozotocin (60 mg/kg) in male Sprague-Dawley rats. One week after streptozotocin injection rats with blood glucose higher than 16.7 mmol/L were taken as diabetics. The study included the following 4 groups: normal control(NC),diabetes mellitus control(DMC),diabetics treated with ultrafine Tongluo recipe (DM+UT,1 g·kg-1·d-1),and diabetics treated with fine Tongluo recipe (DM+FT,1 g·kg-1·d-1).Four weeks after treatment, 24 h urinary protein (24 h UP) and kidney weight/body weight (KW/BW) were detected as renal function indices; malondialdehyde(MDA) and antioxidant enzymes was examined with colorimetry; and NADPH oxidase subunits p47phox and p22phox mRNA were studied with real-time PCR. Results: Compared with the NC group, DMC group had significantly higher MDA,24 h UP, KW/BW, and glutathione peroxidase (GSH-Px) activity, and had significantly lower activity of total superoxide dismutase (TSOD). No significant difference was found in catalase(CAT) activity between the NC group and DMC group. Compared with the 2 Tongluo Recipe groups, the DMC group had significantly lower 24 h UP, KW/BW and MDA, and significantly higher activities of TSOD and GSH-Px, and no significant difference was found in their CAT activities. The effect of ultrafine type on 24 h UP, KW/BW, MDA, TSOD and GSH-Px was significantly greater than the fine type did. The expression of p47phox mRNA of NADPH oxidase in renal cortex group was significantly lower than that in the DMC group, and the latter was significantly higher than the 2 Tongluo of the NC recipe groups, and there was no significant difference between the 2 Tongluo recipe groups. There was no significant difference in p22phox mRNA expression between all the 4 groups.Conclusion: Tongluo recipe may improve the activities of antioxidant enzymes (including TSOD and GSH-Px), decrease p47phox expression, and therefore inhibit the oxidative stress in renal cortex of diabetic rats, reducing the early kidney injury.
Key words:  Tongluo recipe  Diabetes mellitus  Kidney  Antioxidant enzyme and NADPH oxidase