骐龙血脉健方对ApoE^(-/-)小鼠动脉粥样硬化及CD36蛋白表达、Ox-LDL水平的影响

Effects of Qilong Xuemaijian recipe on atherosclerosis,CD36 protein expression and Ox-LDL level in ApoE^(-/-)mice

目的研究骐龙血脉健方对载脂蛋白E基因敲除小鼠(ApoE^(-/-))动脉粥样硬化(AS)模型的血脂水平、主动脉CD36蛋白表达及血清Ox-LDL的影响。方法8周龄健康雄性ApoE^(-/-)小鼠40只,按随机数表法分为模型组、阿托伐组、骐龙血脉健方大、中、小剂量组,每组8只,均给予高脂饮食饲养14周,构建高脂血症小鼠模型。阿托伐组给予阿托伐他汀钙片溶液灌胃,中药干预组分别予骐龙血脉健方颗粒剂大(临床用量20倍)、中(临床用量10倍)、小(临床用量5倍)相应剂量灌胃14周。选取8周龄雄性C57BL/6J野生型小鼠8只作为空白组,给予正常饲料喂养。14周末检测血清总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白(HDL-C)、低密度脂蛋白(LDL-C)水平,ELISA法检测血清氧化低密度脂蛋白(Ox-LDL)水平。分离主动脉,油红染色,检测斑块面积占全动脉内膜面积的比值;蛋白印迹法检测主动脉CD36蛋白表达。结果与空白组相比,模型组TC、TG、LDL-C水平显著升高(P<0.01);与模型组比较,阿托伐组TC、TG、LDL-C水平明显降低(P<0.01),骐龙血脉健方大剂量组TC、TG、LDL-C水平明显降低(P<0.01),骐龙血脉健方中、小剂量组TC水平显著降低(P<0.01),TG、LDL-C降低不明显(P>0.05);大剂量组TC水平低于阿托伐组(P<0.01),而TG水平高于阿托伐组(P<0.05);中、小剂量组TC水平与阿托伐组无明显差异(P>0.05)。与空白组相比,模型组AS斑块面积与内膜总面积比值显著升高(P<0.05);与模型组相比,阿托伐组和骐龙血脉健方大剂量组AS斑块面积与内膜总面积比值明显降低(P<0.05);与阿托伐组相比,大剂量组AS斑块面积与内膜总面积比值无明显变化(P>0.05)。与空白组相比,模型组Ox-LDL水平显著升高(P<0.01);与模型组相比,阿托伐组、骐龙血脉健方大剂量组、中剂量组Ox-LDL水平均降低(P<0.05),其中大剂量组降低最为明显(P<0.01);与阿托伐组相比,大、中剂量组Ox-LDL水平无明显变化(P>0.05)。与空白组相比,模型组主动脉CD36蛋白表达明显升高(P<0.01);与模型组相比,阿托伐组主动脉CD36蛋白表达有所减少(P<0.05),骐龙血脉健方大剂量组主动脉CD36蛋白表达明显减少(P<0.01);与阿托伐组相比,骐龙血脉健方大剂量组主动脉CD36水平无明显变化(P>0.05)。结论骐龙血脉健方可有效改善高脂饮食ApoE^(-/-)小鼠血脂水平,减轻动脉粥样硬化程度,其机制可能与降低血清Ox-LDL水平、下调主动脉CD36蛋白表达有关。

Objective To study the effects of Qilong Xuemaijian recipe on serum lipid levels,aorta CD36 protein expression and serum Ox-LDL in atherosclerotic(AS)mice with apolipoprotein E gene knockout(ApoE^(-/-)).Methods A total of 40 healthy male ApoE^(-/-)mice were divided randomly into five groups:model group,atorvastatin calcium group,Qilong Xuemaijian recipe high,medium and low dose groups,and all mice were given a high-fat diet for 14 weeks to build a hyperlipidemia mouse model.At the same time,8 male C57BL/6J mice aged 8 weeks were selected as blank group and given normal feeding.The mice in atorvastatin calcium group were given atorvastatin calcium solution by gavage.The mice in high,medium and low dose groups were given the corresponding dose of Qilong Xuemaijian recipe.At the end of 14 weeks,the blood of all mice was obtained to detect serum levels of total cholesterol(TC),triglycerides(TG),high density lipoprotein cholesterol(HDL-C),low density lipoprotein cholesterol(LDL-C),and serum oxidized low density lipoprotein(Ox-LDL).The separated aorta was stained with oil red“O”to detect the ratio of atherosclerotic plaque area to total aortic intima area.And the expression of aortic CD36 protein was detected by Western blotting.Results Compared with blank group,the levels of TC,TG and LDL-C were significantly increased in model group(P<0.01).Compared with model group,the levels of TC,TG,LDL-C were significantly reduced in atorvastatin calcium group and high dose group(P<0.01),and TC level was decreased significantly in medium and low dose groups(P<0.01).Compared with atorvastatin calcium group,TC level was decreased in high dose group(P<0.01),while TG level was increased(P<0.05),but there was no significant difference between low,medium dose group and atorvastatin calcium group(P>0.05).Compared with blank group,the ratio of atherosclerotic plaque area to total aortic intima area was increased significantly in model group(P<0.05).Compared with model group,the ratio of atherosclerotic plaque area to total aortic intima area was decreased significantly in atorvastatin calcium group and Qilong Xuemaijian recipe high dose group(P<0.05).There was no difference in the ratio between Qilong Xuemaijian recipe high dose group and atorvastatin calcium group(P<0.05).Compared to blank group,Ox-LDL level was increased significantly in model group(P<0.01).Compared with model group,the Ox-LDL level was decreased significantly in atorvastatin calcium group,Qilong Xuemaijian recipe high and medium dose groups(P<0.05).There was no difference in Ox-LDL level between high,medium dose groups and atorvastatin calcium group(P>0.05).Compared with blank group,the expression of aortic CD36 protein in model group was increased significantly(P<0.01).Compared with model group,the expression of aortic CD36 protein was decreased significantly in atorvastatin calcium group(P<0.05)and Qilong Xuemaijian recipe high dose group(P<0.01),while there was no significant difference in the expression of aortic CD36 protein between Qilong Xuemaijian recipe high dose group and atorvastatin calcium group.Conclusion Qilong Xuemaijian recipe can effectively improve the blood lipid level of ApoE^(-/-)mice fed wtih high-fat diet and reduce the degree of atherosclerosis,which may be related to the inhibition of serum Ox-LDL level and aortic CD36 protein expression.