葛根素降低ApoE-/-小鼠visfatin表达抑制动脉粥样硬化形成的研究

Study of Puerarin on Inhibiting the Progression of Atherosclerosis by Down-regulating Visfatin Expression in Apo E Knockout Mice

目的观察葛根素对载脂蛋白E基因敲除(Apo E-/-)小鼠主动脉粥样硬化(AS)病变、血脂水平、内脏脂肪素(visfatin)及炎症因子IL-6及TNF-α表达的影响。方法 50只SPF级8周龄雄性Apo E-/-小鼠予高脂饲料喂养随机分为5组:葛根素低、中、高剂量组分别给予葛根素300,600,1200 mg·kg-1·d-1灌胃;辛伐他汀组灌服辛伐他汀5 mg·kg-1·d-1;模型组灌服等剂量生理盐水。另设10只相同基因背景和周龄的雄性C57BL/6J小鼠为对照组予普通饲料喂养。16周后测小鼠体质量、血清脂质及visfatin、血浆IL-6及TNF-α含量;HE染色观察小鼠主动脉组织AS斑块面积;western blotting测小鼠主动脉visfatin蛋白的表达。结果各给药组小鼠体质量增加值、血清visfatin、TG、TC、LDL-C水平、血浆IL-6及TNF-α含量、主动脉visfatin蛋白表达均显著低于模型组(P<0.05),血清HDL-C高于模型组(P<0.05);辛伐他汀组、葛根素中、高剂量组小鼠主动脉AS斑块面积百分比显著低于模型组(P<0.05)。结论葛根素可通过降血脂、减小AS斑块面积、降低IL-6及TNF-α含量等发挥抗AS效应,其机制可能与降低visfatin表达有关。

Objective To study the effect of puerarin on serum lipids concentrations,atherosclerotic plaque area quantification at aorta,the circulating levels of serum visfatin and plasma inflammatory cytokines IL- 6 and TNF- α,and visfatin expression in the aorta of Apo E knockout（ Apo E- /-） mice. Methods Fifty male 8- week- old Apo E- /-mice were fed with a western diet（ 21%fat and 0. 15% cholesterol） for 16 weeks to establish atherosclerosis model and randomly assigned into five groups： model group,low- dose puerarin（ 300 mg·kg- 1·d- 1） group,medium- dose puerarin（ 600 mg·kg- 1·d- 1） group,high- dose puerarin（ 1200 mg·kg- 1·d- 1） group and simvastatin（ 5 mg·kg- 1·d- 1） group. Ten C57BL/6J mice fed with a common diet were served as control group. All mice were sacrificed after 16 weeks,serum concentrations of TC,TG,HDL- C,and LDL- C were determined by enzymatic procedures. The circulating levels of serum visfatin,plasma IL- 6 and TNF- α were analyzed using an ELISA method. Atherosclerotic lesions in the aortic root were examined by H E staining. Visfatin expression in the aorta was identified by western blotting. Results The add weight value,serum visfatin,TG,TC and LDL- C,plasma IL- 6 and TNF- α,visfatin expression in the aorta of each treatment group were statistically decreased compared with model group（ P〈0. 05 or P〈0. 01）,while serum HDL- C levels in control group,simvastatin group,high- dose puerarin group and medium- dose puerarin group were higher than that of model group（ P〈0. 05 or P〈0. 01）. Atherosclerotic lesions in the aortic root of simvastatin group,high- dose puerarin group and medium- dose puerarin group were statistically decreased compared with model group（ P〈0. 01）.Conclusion Puerarin can inhibit the progression of atherosclerosis by reducing serum lipids concentrations,atherosclerotic lesions in mice aorta,plasma IL- 6 and TNF- α levels in Apo E- /-mice,the mechanism is related to down- regulating visfatin expression.