艾灸及艾烟对ApoE^(-/-)小鼠血脂、肝脏病理形态及肝脏CD36、ABCA1表达的影响

Effects of Moxibustion and Moxa Smoke on Blood Lipids,and Hepatic Pathologic Morphology and CD36 and ABCA1 Expressions in ApoE^(-/-) mice

目的观察艾灸及艾烟干预对动脉粥样硬化模型血脂、肝脏病理改变和肝细胞内胆固醇代谢相关分子的影响,分析艾灸和艾烟对胆固醇代谢的调节作用,探讨艾灸及艾烟的作用机制。方法将51只8周龄ApoE^(-/-)小鼠随机分为模型组、艾烟组、艾灸组,每组17只,20只C57BL/6小鼠作为空白对照。正常组、模型组小鼠常规抓取和固定;艾烟组小鼠暴露于10 mg/m^3~15 mg/m^3艾烟环境;艾灸组小鼠艾灸关元穴。所有干预每日20 min,每星期6次,连续干预12星期。全自动生化分析仪检测总胆固醇(TC)、甘油三酯(TG)和高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C);HE染色观察肝脏病理形态;免疫组化法检测肝脏CD36、ABCA1的表达。结果模型组小鼠血清TG、LDL-C含量显著高于正常组(P=0.003,P=0.001),HDL-C含量显著低于正常组(P=0.007),TC含量与正常组无显著差异(P>0.05);艾灸组小鼠血清TG、LDL-C含量显著低于模型组(P=0.03,P=0.001),HDL-C、TC含量与模型组无显著差异(P=0.11,P=0.11);艾烟组小鼠血清TG、LDL-C含量显著低于模型组(P=0.01,P=0.008),HDL-C含量与模型组无显著差异(P>0.05);艾灸组与艾烟组各项血脂指标均无显著性差异(P>0.05)。模型组小鼠肝细胞索、肝窦排列紊乱,肝细胞肿胀变性明显。艾烟组、艾灸组小鼠肝细胞肿胀程度明显减轻,炎细胞浸润程度较模型组减轻。模型组CD36表达显著高于正常组(P=0.004),ABCA1表达显著低于正常组(P<0.001);艾灸组CD36表达与模型组无显著性差异(P=0.09),ABCA1表达显著高于模型组(P=0.03);艾烟组CD36表达显著低于模型组(P=0.02),ABCA1表达显著高于模型组(P=0.002);艾灸组与艾烟组CD36、ABCA1表达均无显著差异(P>0.05)。结论早期艾灸关元穴可以一定程度调整动脉粥样硬化模型ApoE^(-/-)小鼠血脂代谢紊乱的状态,减缓肝脏病变发生,减少肝脏内胆固醇的蓄积,这可能是艾灸疗法预防动脉粥样硬化的作用机制之一。艾烟作为艾灸生成物,是艾灸发挥治疗作用的有效因素。

Objective To investigate the intervention effects of moxibustion and moxa smoke on blood lipids,hepatic pathological changes and intrahepatocytic molecules related to cholesterol metabolism and analyze the regulating effects of moxibustion and moxa smoke on cholesterol metabolism and explore the mechanisms of actions of moxibustion and moxa smoke.Method Fifty-one 8-week-old ApoE-（-/-） mice were randomized into model,moxa smoke and moxibustion groups,17 mice each.Twenty C57BL/6 mice comprised a blank control group.The normal and model groups of mice were routinely grabbed and fastened.The moxa smoke group of mice was exposed to 10-15 mg/m-3 moxa smoke circumstances.The moxibustion group of mice was given moxibustion on point Guanyuan（CV4）.All interventions were made 20 min daily,6 times a week,for 12 consecutive weeks.Total cholesterol（TC）,triglyceride（TG）,high density lipoprotein-cholesterol（HDL-C） and low density lipoprotein-cholesterol（LDL-C） were measured using an automatic biochemical analyzer.Hepatic pathologic morphology was observed by HE staining.Hepatic CD36 and ABCA1 expressions were determined by immunohistochemical method.Result In the model group of mice,serum TG and LDL-C contents were significantly higher than in the normal group（P=0.003;P=0.001）;HDL-C content was significantly lower than in the normal group（P=0.007）;TC content had no significant difference compared with the normal group（P〈0.05）.In the moxibustion group of mice,serum TG and LDL-C contents were significantly lower than in the model group（P=0.03;P=0.001） and HDL-C content had no significant difference compared with the model group（P=0.11）.In the moxa smoke group of mice,serum TG and LDL contents were significantly lower than in the model group（P=0.01;P=0.008） and HDL content had no significant difference compared with the model group（P=0.11）.There were no significant differences in various blood lipid indicators between the moxibustion and moxa smoke groups（P〈0.05）.There were hepatic cell cord and sinusoid derangement and obvious hepatocytic swelling in the model group of mice.In the moxa smoke and moxibustion groups,hepatocytic swelling subsided significantly,and inflammatory cell infiltration reduced compared with the model group.In the model group,CD36 expression was significantly higher than in the normal group（P=0.004） and ABCA1 expression was significantly lower than in the normal group（P=0.001）.In the moxibustion group,CD36 expression had no significant difference compared with the model group（P=0.09） and ABCA1 expression was significantly higher than in the model group（P=0.03）.In the moxa smoke group,CD36 expression was significantly lower than in the normal group（P=0.02） and ABCA1 expression was significantly higher than in the model group（P=0.002）.There were no significant differences in CD36 and ABCA1 expressions between the moxibustion and moxa smoke groups（P〈0.05）.Conclusion Early moxibustion on point Guanyuan can regulate disorders of blood lipid metabolism,delay the occurrence of hepatic lesions and reduce intrahepatic accumulation of cholesterol to a certain extent in an ApoE-（-/-） mouse model of atherosclerosis.That may be one of the mechanisms by which moxibustion therapy prevents atherosclerosis.Moxa smoke as the product of moxibustion is an effective factor in moxibustion producing a therapeutic effect.