隔药饼灸对动脉粥样硬化兔血浆三甲胺、氧化三甲胺及肝黄素单加氧酶3的影响

Effects of herbal cake-partitioned moxibustion on plasma trimethylamine,trimethylamine-N-oxide,and flavin-containing monooxygenase 3 in atherosclerotic rabbits

目的观察隔药饼灸对动脉粥样硬化(AS)兔血浆三甲胺(TMA)、氧化三甲胺(TMAO)水平以及肝黄素单氧化酶3(FMO3)含量的影响,探讨隔药饼灸治疗AS的可能机制。方法将28只雄性新西兰兔适应性喂养1周后根据随机数字表法分为空白组、模型组、抗生素组和隔药饼灸组,每组7只。空白组予基础饲料喂养,其余各组予基础饲料加1%胆碱喂养,制备AS模型。抗生素组予添加广谱抗生素的饮水,隔药饼灸组予隔药饼灸,持续12周。干预结束后,采用苏木素-伊红(HE)染色观察动脉粥样硬化斑块,检测各组兔血脂水平,液相色谱-质谱联用检测血浆TMA及TMAO含量。免疫印迹法检测肝脏FMO3蛋白表达。实时荧光定量聚合酶链反应法检测肝脏FMO3 mRNA表达。结果HE染色结果显示模型组动脉管壁粗糙,内膜明显增厚,可见较多泡沫细胞、脂质沉积;隔药饼灸组动脉壁增厚不明显,有少量泡沫细胞和脂质沉积。与空白组比较,模型组血清中低密度脂蛋白胆固醇(LDL-C)、总胆固醇(TC)水平升高(P<0.01),血浆中TMA及TMAO水平升高(P<0.05,P<0.01),肝脏FMO3蛋白及mRNA表达升高(P<0.05,P<0.01);血清高密度脂蛋白胆固醇(HDL-C)水平下降(P<0.05)。与模型组比较,隔药饼灸组及抗生素组LDL-C及TC水平下降(P<0.01或P<0.05),HDL-C水平上升(P<0.01),TMA及TMAO水平下降(P<0.05),FMO3蛋白及mRNA表达下降,但差异无统计学意义。结论隔药饼灸能减缓动脉粥样硬化斑块形成,调节AS兔的血脂水平,其机制可能与下调血浆中TMA及TMAO表达相关。

Objective To investigate the effects of herbal cake-partitioned moxibustion on the plasma levels of trimethylamine(TMA),trimethylamine-N-oxide(TMAO),and flavin-containing monooxygenase 3(FMO3)in rabbits with atherosclerosis(AS),as well as to explore the possible mechanism of herbal cake-partitioned moxibustion in treating AS.MethodsAfter 1-week adaptive feeding,28 male New Zealand rabbits were divided into a blank group,a model group,an antibiotic group,and a herbal cake-partitioned moxibustion group according to the random number table method,with 7 rabbits in each group.Rabbits were fed with a basic diet in the blank group,while with a basic diet plus 1%choline in the remaining groups to prepare the AS model.Rabbits were given drinking water with broad-spectrum antibiotics in the antibiotic group,and herbal cake-partitioned moxibustion in the herbal cake-partitioned moxibustion group for 12 weeks.The atherosclerotic plaques by hematoxylin-eosin(HE)staining,the blood lipid levels,the plasma TMA and TMAO levels by liquid chromatography-mass spectrometry were detected for rabbits in each group at the end of interventions.Liver FMO3 protein expression was detected by Western blotting.Liver FMO3 mRNA expression was detected by real-time fluorescence quantitative polymerase chain reaction.ResultsHE staining showed that the arterial wall was rough,the intima was significantly thickened,and more foam cells and lipid deposits were seen in rabbits of the model group.Arterial wall thickening was not obvious with a few foam cells and lipid deposits in the herbal cake-partitioned moxibustion group.Compared with the blank group,the serum levels of low-density lipoprotein cholesterol(LDL-C)and total cholesterol(TC)were increased(P<0.01),the plasma levels of TMA and TMAO were increased(P<0.05,P<0.01),and the expression levels of liver FMO3 protein and mRNA were all increased(P<0.05,P<0.01);while the serum high-density lipoprotein cholesterol(HDL-C)level was decreased in the model group(P<0.05).Compared with the model group,the LDL-C and TC levels were decreased(P<0.01 or P<0.05),the HDL-C levels were increased(P<0.01),the TMA and TMAO levels were decreased(P<0.05),while the protein and mRNA expression levels of FMO3 were decreased without statistical significance in the herbal cake-partitioned moxibustion group and the antibiotic group.ConclusionHerbal cake-partitioned moxibustion can slow atherosclerotic plaque formation and regulate lipid levels in AS rabbits,and the mechanism may be related to the down-regulation of TMA and TMAO expression in the plasma.