白头翁皂苷B4对高脂饮食诱导ApoE^(-/-)小鼠动脉粥样硬化的治疗作用及机制研究

Therapeutic effect and mechanism of Pulsatilla saponin B4 on atherosclerosis of ApoE^(-/-) mice induced by high fat diet

目的研究白头翁皂苷B4的抗动脉粥样硬化作用及其机制。方法将雄性的C57小鼠设为对照组,共6只,给予正常饮食;给予ApoE^(-/-)小鼠高脂饮食喂养9周后,随机分为4组:模型组、辛伐他汀(阳性对照,5 mg·kg^(-1))组及白头翁皂苷B4低、高剂量(5、20 mg·kg^(-1))组,每组7只。ig给药9周,C57对照组和ApoE^(-/-)模型组给予等量的0.9%的氯化钠溶液。给药的同时造模小鼠仍给予高脂饮食,每周统计1次体质量,给药9周后进行取材。通过兽用血常规分析仪检测全血中血小板(PLT)、白细胞(WBC)、中性粒细胞(Neu)及淋巴细胞(Lym)水平;ELISA法检测血清中白细胞介素-6(IL-6)的水平;试剂盒法检测血清中总胆固醇(TC)、游离胆固醇(FC)、三酰甘油(TG)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)水平;通过全长主动脉油红染色检测主动脉中脂质积累情况;通过苏木精-伊红染色法、Masson染色法和油红O染色法检测主动脉窦的组织病理变化。结果与模型组比较,在白头翁皂苷B4及辛伐他汀给药后,体质量呈降低趋势;PLT、WBC、Neu、Lym、IL-6水平均显著降低(P<0.05、0.01、0.001);血清TC、FC、TG、LDL-C水平显著降低(P<0.001),HDL-C水平显著升高(P<0.001);全长主动脉被染红区域显著缩小(P<0.001);主动脉窦油红染色区域面积明显减少,小鼠主动脉窦斑块减少,可见少量泡沫细胞,炎症细胞浸润减少,中膜层平滑肌及弹性纤维排列增厚不明显,胶原含量多且分布规律,胶原间的腔隙减少,油红染色区域面积明显减少。结论白头翁皂苷B4对高脂饮食诱导的ApoE^(-/-)基因敲除小鼠动脉粥样硬化具有治疗作用,其作用机制与减少脂质浸润和炎症反应有关。

Objective To study the anti atherosclerosis effect of Pulsatilla saponin B4 and its mechanism.Methods Male C57 mice were set as the control group,with a total of six mice,and were given a normal diet.After nine weeks of high-fat diet feeding,ApoE^(-/-)mice were randomly divided into four groups:the model group,the simvastatin(positive control,5 mg·kg^(-1))group,and the Pulsatilla saponin B4 low-and high-dose(5 and 20 mg·kg^(-1))groups,with seven mice in each group.The mice were ig administered with the drugs for nine weeks,and the C57 control group and the ApoE^(-/-)model group were given an equal amount of 0.9%sodium chloride solution.While the treated mice were still given a high-fat diet,their body weights were measured once a week,and tissue samples were collected after nine weeks of treatment.The levels of platelets(PLT),white blood cells(WBC),neutrophils(Neu),lymphocytes(Lym),and interleukin-6(IL-6)in the whole blood were detected by veterinary hematology analyzer;the level of IL-6 in the serum was detected by ELISA,the levels of total cholesterol(TC),free cholesterol(FC),triglycerides(TG),low-density lipoprotein cholesterol(LDL-C),and high-density lipoprotein cholesterol(HDL-C)in the serum were detected by kit method,the accumulation of lipid in the aorta was detected by whole aorta oil red staining,the histopathological changes of the aortic sinus were detected by hematoxylin and eosin staining,Masson staining,and oil red O staining.Results Compared with the model group,there was a trend of body weight reduction in the Pulsatilla saponin B4 and simvastatin treatment groups,the levels of PLT,WBC,Neu,Lym,and IL-6 were significantly lower(P<0.05,0.01,0.001).The levels of TC,FC,TG,and LDL-C in serum were significantly lower(P<0.001),while the level of HDL-C was significantly higher(P<0.001).The area of the red-stained entire aorta was significantly reduced(P<0.001),the area of the oil-red-stained aortic sinus was significantly reduced,the aortic plaque in mice was reduced,and there were only a few foam cells visible,with a reduction in inflammatory cell infiltration,thickening of the smooth muscle and elastic fibers in the media layer was not obvious,the collagen content was abundant and distributed regularly,the interstitial spaces between collagen were reduced,and the area of the oil-red-stained region was significantly reduced.Conclusion Pulsatilla saponin B4 had therapeutic effect on atherosclerosis of ApoE^(-/-)mice induced by high fat diet and its mechanism was related to reducing paper infiltration and inflammatory response.