小檗皮水浸膏对db/db糖尿病小鼠视网膜病变的影响(Ⅱ)  

Effect of Water Extract from Cortex of Berberis dictyophylla on Retinopathy in Spontaneous Type 2 Diabetic db/db Mice( Ⅱ)

作　　者：周邦华[1] 叶凡[1] 岳丽珺 孟宪丽[1] 范刚[1] 赖先荣[1] 

机构地区：[1]成都中医药大学,成都611137

出　　处：《中国实验方剂学杂志》2016年第24期116-121,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81173360;81303310;81473427;81560806);四川省科技厅科技支撑计划项目(2011SZ0298);四川省教育厅重点项目(10ZA092);四川省高校科研创新团队建设计划项目(11TD004)

摘　　要：目的:观察小檗皮浸膏对基因突变自然发病型db/db糖尿病小鼠视网膜蛋白激酶C-β(PKC-β),缺氧诱导因子-1α(HIF-1α),血管内皮细胞生长因子(VEGF)的影响,探索该药防治糖尿病视网膜病变的作用机制。方法:选用18周龄db/db糖尿病小鼠,db/m型同性同窝瘦型小鼠为对照。采用随机方法将db/db小鼠分为db/db组,小檗皮高、中、低剂量组(1.50,0.75,0.38 g·kg^(-1)),羟苯磺酸钙组(0.23 g·kg^(-1)),盐酸小檗碱组(0.135 g·kg^(-1))。相关药物连续ig 2个月,并于ig 2个月末处死小鼠。采用免疫组化方法观察3种因子在视网膜结构的表达情况,实时荧光定量聚合酶链式反应(Real-time PCR)和蛋白质免疫印迹(Western blot)方法检测视网膜内3种因子的基因与蛋白表达。结果:与db/m组比较,db/db组视网膜组织PKC-β,VEGF免疫组化反应呈阳性,表达显著增加(P<0.01);HIF-1α免疫组化反应呈阳性,表达量有升高趋势,但差异无统计学意义;db/db组三种因子的基因与蛋白表达显著增加(P<0.01)。与db/db组比较,羟苯磺酸钙组视网膜组织PKC-β免疫组化反应呈阳性,表达显著降低(P<0.05),小檗皮高剂量组视网膜组织PKC-β免疫组化反应呈阳性,表达显著降低(P<0.01),其他各给药组较db/db组PKC-β表达有降低趋势,但差异无统计学意义;各给药组视网膜组织HIF-1α免疫组化反应呈阳性,表达有降低趋势,但差异无统计学意义;小檗碱组、小檗皮低剂量组视网膜组织VEGF免疫组化反应呈阳性,表达明显降低(P<0.01),其他各给药组视网膜组织VEGF有降低趋势,但差异无统计学意义。与db/db组比较,羟苯磺酸钙组HIF-1α基因表达明显降低(P<0.05),其他各给药组PKC-β,HIF-1α,VEGF基因表达有降低趋势,但差异无统计学意义。与db/db组比较,羟苯磺酸钙组、盐酸小檗碱组、小檗皮高、低剂量组PKC-β蛋白表达显著降低(P<0.01),小檗皮中剂量组PKC-β蛋白表达有降低趋势,但差异无统计学Objective： To observe the effects of water extract from cortex of Berberis dictyophylla（ BDCE） on expressions of retinal protein kinase C-β（ PKC-β）,hypoxia inducing factor-1α（ HIF-1α）,vascular endothelial growth factor（ VEGF） in spontaneous type 2 diabetic db / db mice,in order to explore its mechanism in preventing and treating diabetic retinopathy（ DR）. Method： Eighteen-week-old db / db mice were selected,with their thin db / m mice for normal control group. The db / db mices were randomly divided into 6 groups： model control（ db / db） group,high,middle and low-dose BDCE group（ 1. 50,0. 75,0. 38 g·kg^（-1））,calcium-dobesilate group（ 0. 23 g·kg^（-1））,and berberine hydrochloride group（ 0. 135 g·kg^（-1））. The mice were given relevant drugs for consecutively two months. At the end of the two-month treatment,all the mice were put to death. The immunohistochemical method was used to observe the expressions of the three factors on the retina. Real-time quantitative PCR and western blot methods were used to detect genes and proteins expressions level of PKC-β,HIF-1α and VEGF in retina. Result： Compared to db / m group,the immunohistochemical results showed positive reaction of PKC-β and VEGF in retinal tissues of db / db group,with significant increase in expressions（ P〈0. 01）; positive reaction of HIF-1α,with no significant increase in expressions of db / db group; mRNA and protein expressions of PKC-β,HIF-1α,VEGF increased significantly in db / db group（ P〈0. 01）. Compared to db / db group,the immunohistochemical results showed positive reaction of PKC-β on retinal tissues in calcium-dobesilate group,with significant decrease of expression（ P〈0. 05）; positive reaction of PKC-β on retinal tissues in highdosage BDCE group,with significant decrease of expression（ P〈0. 01）; a downward trend in PKC-β expression in other treatment groups compared with the db / db group,with no statistical significance in difference; positive reac

关 键 词：小檗皮 基因突变自然发病型db/db糖尿病小鼠 糖尿病视网膜病变 蛋白激酶-β 缺氧诱导因子-1Α 血管内皮细胞生长因子 

分 类 号：R285.5[医药卫生—中药学]