补阳还五汤加味预防动脉粥样硬化形成机制

目的:探究补阳还五汤加味对动脉粥样硬化(AS)形成的预防及机制研究。方法:将30只雄性载脂蛋白E基因敲除小鼠(Apo E-/-小鼠)随机分为Apo E-/-小鼠组、模型组、补阳还五汤加味组和辛伐他汀组,另以正常雄性C57BL/6J小鼠设组。模型组、补阳还五汤加味组和辛伐他汀组采用高脂饮食造模。第4周后,补阳还五汤加味组和辛伐他汀组分别灌以补阳还五汤20 g·kg~(-1)+水蛭粉0.46 g·kg~(-1),辛伐他汀3 mg·kg~(-1)。第9周后,检测血清总胆固醇(TC),甘油三酯(TG),高密度脂蛋白胆固醇(HDL-C)和低密度脂蛋白胆固醇(LDL-C)值,取动物主动脉根部经苏木素-伊红(HE)常规染色后观察组织形态变化并测量纤维帽厚度和内-中膜厚度、检测主动脉p38丝裂原活化蛋白激酶(p38MAPK)和细胞外信号调节激酶5(ERK5)蛋白表达变化。结果:血脂水平,与C57BL/6J小鼠正常组比较,Apo E-/-小鼠组和模型组TC,TG,LDL-C均升高且HDL-C均降低(P0.01);与模型组比较,补阳还五汤加味组与辛伐他汀组均能降低Apo E-/-小鼠TC,TG,LDL-C水平,同时升高HDL-C(P0.05,P0.01)。HE染色,C57BL/6J小鼠正常组主动脉管壁无斑块形成,Apo E-/-小鼠组管壁有极少斑块形成,内-中膜略有增厚。模型组斑块形成明显,内-中膜明显增厚。补阳还五汤加味组与辛伐他汀组较模型组斑块减少,内-中膜厚度减少。蛋白表达,与C57BL/6J小鼠正常组比较,模型组主动脉组织中p38MAPK的蛋白表达量增多(P0.01),ERK5的蛋白表达量减少(P0.01);与模型组比较,补阳还五汤加味组和辛伐他汀组均能降低p38MAPK蛋白的表达量(P0.01),增加组织中ERK5蛋白的表达(P0.01)。结论:补阳还五汤加味与辛伐他汀均有预防AS的作用。补阳还五汤加味预防AS发生的机制可能与干预MAPK信号通路中的p38MAPK,ERK5途径有关。

Preventive Effect of Modified Buyang Huanwu Tang on Formation of Atherosclerosis

Objective: To study the preventive mechanism of modified Buyang Huanwu Tang on formation of atherosclerosis. Method: Thirty male ApoE^-/- mice were randomly divided into four groups:ApoE^-/- mice blank group, model group, modified Buyang Huanwu Tang group and simvastatin group. Normal male C57BL/6J mice were used as a normal group. High-fat diet was given for modeling in model group, modified Buyang Huanwu Tang group and simvastatin group. 4 weeks later, the mice in Buyang Huanwu Tang group or simvastatin group were intragastrically administrated with Buyang Huanwu Tang (20 g·kg^-1)+leech powder (0.46 g·kg^-1) or simvastatin(3 mg·kg^-1). After 9 weeks, the total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-C) in serum were detected. In addition, aorta segment was taken from the experiment animals to observe morphological changes by HE staining,measure fiber-cap thickness and vascular intima-media thickness. Western blot was used to determine the expression levels of aortic p38 mitogen activated protein kinase(p38MAPK) and extracellular signal-regulated kinase 5(ERK5). Result: As compared with the normal group, TC, TG and LDL-C levels were significantly increased in the ApoE^-/-mice blank group and model group, while HDL-C level was significantly decreased (P<0.01). As compared with the model group, TC, TG and LDL-C levels were decreased in the modified Buyang Huanwu Tang group and simvastatin group, while HDL-C level was significantly increased (P<0.05, P<0.01). HE staining showed no aortic plaque formation in C57BL/6J mice normal group, few aortic plaque formation and slightly increased vascular intima-media thickness in ApoE^-/- mice blank group, significant aortic plaque formation and increased vascular intima-media thickness in model group. As compared with the model group, aortic plaque formation and vascular intima-media thickness was reduced in both modified Buyang Huanwu Tang group and simvastatin group. The protein expression level of aortic p38MAPK in model group was higher while protein expression level of ERK5 was lower than those in C57BL/6J normal group (P<0.01). In addition, the protein expression level of aortic p38MAPK was significantly decreased while the protein expression level of ERK5 was increased in both modified Buyang Huanwu Tang group and simvastatin group as compared with model group (P<0.01, P<0.01). Conclusion: Both modified Buyang Huanwu Tang and simvastatin have an effect in preventing the development of atherosclerosis, and the potential mechanism could be related to intervening p38MAPK and ERK5 in MAPK signal pathway.