原花青素对I型糖尿病大鼠血管内皮功能、氧化性损伤和凋亡相关蛋白表达的影响
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The Influence of Procyanidins on Function,Oxidative Damage and Apoptosis Related Protein Expression of Vascular Endothelial Cell in Type I Diabetic Rats
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    目的 研究原花青素(procyanidins,PC)对糖尿病大鼠血管内皮细胞的作用,并在三种凋亡相关蛋白水平探讨作用机制。方法 雄性SD大鼠,尾静脉注射链脲佐菌素(STZ)30 mg/kg造成1型糖尿病动物模型,随机分为糖尿病模型组,50 mg/kg原花青素治疗组和100 mg/kg原花青素治疗组,每组8只,另选8只健康雄性大鼠为正常对照组。各组大鼠灌服给药12周后,用血糖仪及专用血糖试纸现场检测各组大鼠空腹血糖;采用放射免疫分析仪检测血清内皮素1(ET-1)和前列环素(PGI2)水平;采用比色法检测血管内皮组织MDA含量和SOD活性;采用常规免疫组织化学染色方法对血管内皮细胞三种凋亡相关蛋白Caspase-3、Bax和Bcl-2的表达进行检测;用Olympus成像分析系统高倍镜(×400)下观察和采集显微图像,采用美国显微镜图像分析测量软件Image-Pro Plus 6.0采集整个视野黄染灰度值并自动转化为光密度值,计算出单个内皮细胞平均光密度值(mean-density),分析三种凋亡相关蛋白的表达情况。结果 糖尿病模型组大鼠,与正常对照组比较,大鼠空腹血糖水平明显增高,血清ET-1含量增高和PGI2含量降低(P<0.01),血管内皮组织MDA含量提高(P<0.01),血管内皮组织SOD活性降低(P<0.01),血管内皮Caspase-3 和 Bax蛋白表达均增强(P<0.01),Bcl-2 蛋白表达及Bcl-2和Bax蛋白表达比值(Bcl-2/Bax)均减弱(P<0.01);50 mg/kg原花青素治疗组和100 mg/kg原花青素治疗组,与糖尿病模型组比较,大鼠空腹血糖水平不同程度减低(P<0.05和P<0.01),血清ET-1含量降低和PGI2含量升高(P<0.01),血管内皮组织MDA含量降低(P<0.01),血管内皮组织SOD活性提高(P<0.05和P<0.01),血管内皮Caspase-3 和 Bax蛋白表达均减弱(P<0.01),Bcl-2 蛋白表达及Bcl-2和Bax蛋白表达比值(Bcl-2/Bax)均提高(P<0.01),SOD含量均高于糖尿病模型组。结论 原花青素对糖尿病大鼠血管内皮分泌功能和氧化性损伤有调节保护作用,同时可抑制血管内皮Caspase-3和Bcl-2蛋白表达水平,促进Bcl-2表达,提高Bcl-2和Bax蛋白表达比值。

    Abstract:

    Aim To investigate the effect of Procyanidins on vascular endothelial cell in diabetic rats and discuss the possible mechanism. Methods Male SD rats were injected streptozotocin ( STZ, 30 mg/k g ) by tail intravenous for establishing the type I diabetes animal model,randomly divided into diabetes model group,50 mg/kg procyanidins treatment group and 100 mg/kg procyanidins treatment group,and other 8 healthy male SD rats were included into the control group. The rats took medicine by irrigation stomach for 12 weeks,rat fasting blood glucose was detected with glucose meter and special in-situ;serum ET-1 and PGI2 level were detected with radiation immunity analyzer;MDA and SOD in vascular endothelial cell were detected by the methods of ELISA Caspase-3 ,Bax and Bcl-2 in vascular endothelial cell were detected by the conventional immunohistochemical treatment and dyeing methods;vascular endothelial cell microscopic image was observed and collected with being future-proof imaging analysis system (× 400) at high magnification;Caspase-3 ,Bax and Bcl-2 in vascular endothelial cell were analysed by mean-density using the microscope Image analysis software Image - Pro Plus 6.0 in yellow. Results Compared with the normal control group,in Diabetes model rats,rat fasting blood glucose level increased, serum ET-1 level increased and PGI2 level reduced (P< 0.01),MDA in vascular endothelial cell increased(P< 0.01),SOD in vascular endothelial cell reduced(P< 0.01),Caspase-3 and Bax in vascular endothelial cell increased(P< 0.01) Bcl-2 and Bcl-2/Bax in vascular endothelial cell reduced(P< 0.01)In 50 mg/kg procyanidins treatment group and 100 mg/kg procyanidins treatment group,rat fasting blood glucose level decreased (P< 0.05,P< 0.01),serum ET-1 level reduced and PGI2 level increased(P< 0.01),MDA in vascular endothelial cell reduced (P< 0.01),SOD in vascular endothelial cell increased (P< 0.05,P< 0.01),Caspase-3 and Bax in vascular endothelial cell reduced (P< 0.01) Bcl-2 and Bcl-2/Bax in vascular endothelial cell increased (P< 0.01). Conclusion Procyanidins has protective effect on diabetic rats vascular endothelial cell secretion function and oxidative damage,meanwhile can inhibit Caspase-3 and Bax ,induce Bcl-2 and Bcl-2/Bax of diabetic rat vascular endothelial cell.

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姜志云,刘 义.原花青素对I型糖尿病大鼠血管内皮功能、氧化性损伤和凋亡相关蛋白表达的影响[J].中国动脉硬化杂志,2015,23(03):271~276.

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  • 收稿日期:2014-08-06
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