菲牛蛭素对载脂蛋白E基因敲除小鼠动脉粥样硬化斑块的干预作用

Interventional effects of Bufrudin on atherosclerosis plaque in Apo E gene knockout mice

目的研究菲牛蛭素对载脂蛋白E基因敲除(ApoE^(-/-))小鼠动脉粥样硬化及斑块的干预作用。方法按照体重将高脂饲料喂养的雄性ApoE^(-/-)小鼠随机分为3组:模型组、对照组(给予辛伐他汀3mg·kg^(-1)·d^(-1)灌胃)和实验组(给予菲牛蛭素400 U·kg^(-1)·d^(-1)腹腔注射);另设C57BL/6J小鼠为正常组(不予任何处理)。给药10周后,全自动生化仪检测各组小鼠血清三酰甘油(TG)、总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)及高密度脂蛋白胆固醇(HDL-C)的水平,以免疫组化法检测主动脉血管标记物CD34蛋白、血管内皮生长因子(VEGF)及血管内皮生长因子受体(VEGFR-2)的表达。结果给药后,模型组、对照组和实验组血清TG分别为(1.57±0.12),(1.10±0.21),(0.95±0.20)mmol·L^(-1);这3组的TC分别为(19.93±2.24),(14.47±1.30),(11.90±2.38)mmol·L^(-1);这3组的LDL-C分别为(17.24±1.52),(13.94±1.46),(10.02±2.14)mmol·L^(-1);这3组的HDL-C分别为(4.05±1.81),(4.62±0.50),(3.38±0.24)mmol·L^(-1),与模型组相比,实验组差异均有统计学意义(均P<0.01)。模型组、对照组和实验组的斑块胶原相对含量分别为(21.48±2.44)%,(37.12±3.33)%,(51.60±2.47)%;这3组的CD34蛋白含量分别为(11.02±1.64)%,(6.82±0.66)%,(4.56±0.59)%;这3组的VEGF蛋白分别为(22.13±2.04)%,(16.20±0.65)%,(11.16±0.82)%;这3组的VEGFR-2蛋白表达分别为(17.90±0.73)%,(15.11±0.52)%,(10.56±0.60)%;与模型组相比,实验组差异均有统计学意义(均P<0.01)。结论菲牛蛭素可改善ApoE^(-/-)小鼠动脉粥样硬化,稳定粥样斑块,其机制可能与减少VEGF和VEGFR-2表达、抑制斑块微血管新生有关。

Objective To study the interventional effect of Bufrudin on aorta atherosclerosis and angiogenesis in apolipoprotein E gene knockout （ApoE-/- ） mice. Methods Male ApoE-/- mice on high fat diet, were randomly divided into three groups ：model group, control group, experimen- tal group. Experimental group were given Bufrudin （400 U · kg-1 . d-1 ）, control group were given simvastatin （3 mg · kg-1 · d-1）. And C57BL/ 6J mice fed with common diet was served as normal group. After ten weeks, mice blood was colleeted and serum levels of triglyeeride （TG） , total cholesterol （ TC）, low - density lipoprotein cholesterol （ LDL- C ）, high density lipoprotein cholesterol （HDL - C ） concentrations weremeasured. The expression of CD34 protein,vascular endothelial growth factor （VEGF） and vascular endothelial growth factor receptor 2 （VEGFR -2 ） were detected by immunohistochemstry. Results After administraiton, the serum content of TG in model group, control group and experimental group were （ 1.57 ±0. 12 ）, （ 1.10 ± 0.21 ）, （0.95±0.20） mmol · L^-l,respectively;TC in above three groups were （19.93 ± 2. 24）, （14.47 ± 1,30）, （11.90±2.38） mmol. L-1 respectively; LDL-Cinabovethree groups were （17.24 ±1.52） （13.94 ±1.46） （10.02±2. 14） mmol · L-1 respectively;HDL-C in above three groups were （4.05 ±1.81） （4.62±0.50） （3.38 ±0. 24） mmol · L- 1, respectively. Compared with model group, the serum content of TG, TC, LDL - C, HDL - C in experimental group were significantly decreased（ all P 〈0. 01 ）. The content of collagen in model group ,control group and experimental group were （21.48 ±2. 44 ） %, （ 37.12 ±3.33 ） %, （51.60 ± 2.47 ） % ; the expression of CD34 protein in above three groups were （ 11.02 ± 1.64 ） % , （ 6. 82 ± 0.66 ） % , （ 4. 56 ± 0. 59 ） % ; the expression of VEGf prtein in above three groups were （ 22.13 ±2. 04 ） %, （ 16.20 ± 0.65 ） %, （ 11.16 ± 0.82 ） % ; and the expression of VEGfR -2 in above three groups were （ 17.90 ±0. 73） %, （ 15.11 ±0. 52） % , （ 10. 56 ±0. 60） % ,respectively. Com- pared with model group, the content of collagen in experimental group was increased, and the expression of CD34 protein in experimental group was decreased, all about with statistical difference （ all P 〈 0.05 ） . Conclusion Bufrudin attenuate atheroselerosis disease and stabilize the atherosclerotic plaque in ApoE -/- mice likely through suppressing the expression of VEGF and VEGFR -2.