基于代谢组学研究埋线丰隆及其配穴对NAFLD大鼠的协同和拮抗作用

Synergistic and Antagonistic Effects of Catgut Embedding at ST40 and Its Matching Points in NAFLD Rats Based on Metabolomics

目的:观察埋线丰隆穴、丰隆加足三里穴和丰隆加三阴交穴对高脂饲料(HFD)诱导的非酒精性脂肪性肝病(NAFLD)模型大鼠血脂4项(TG、TC、LDL-C及HDL-C)、肝脏病理及血清代谢物的影响。方法:将50只SD大鼠随机分为正常组、模型组、丰隆组、丰隆加足三里组和丰隆加三阴交组,每组10只。采用HFD喂养制备NAFLD模型大鼠,正常组和模型组大鼠不做任何干预,埋线组分别给予大鼠丰隆穴、丰隆加足三里穴和丰隆加三阴交穴进行埋线治疗,每周埋线1次,干预4周,造模同时给予埋线治疗。采用全自动生化分析仪检测血清TC、TG、HDL-C和LDL-C水平,用HE染色及油红染色观察大鼠肝脏病理变化,用非靶代谢组学分析各组大鼠血清代谢物变化。结果:与正常组比较,模型组血清TC、TG及LDL-C水平升高(P<0.01),血清HDL-C水平下降(P<0.01),肝脏脂肪、脂滴增加。与模型组比较,丰隆组和丰隆加三阴交组血清TC、TG及LDL-C水平降低(P<0.01),而丰隆加足三里穴组血清TC、TG及LDL-C水平无明显降低,差异无统计学意义(P>0.05),3组埋线组大鼠肝脏脂肪堆积均具有一定程度的改善,丰隆加三阴交组改善程度最为显著,丰隆组次之,丰隆加足三里穴组改善效果最差。丰隆加三阴交组和丰隆组对胆碱代谢通路影响较为显著,差异有统计学意义(P<0.01),但丰隆加足三里组对此通路无明显影响,差异无统计学意义(P>0.05)。结论:埋线丰隆穴及丰隆加三阴交穴均可改善NAFLD大鼠血脂水平,但丰隆加足三里未能显著改善NAFLD大鼠血脂水平,提示丰隆配伍三阴交对NAFLD大鼠具有协同作用,但丰隆配伍足三里可能具有一定的拮抗作用,代谢组学揭示了胆碱代谢通路可能是其协同和拮抗作用的内在机制。

Objective:To observe the effects of catgut embedding at Fenglong(ST40),ST40 plus Zusanli(ST36) and ST40 plus Sanyinjiao(SP6) on TG,TC,LDL-C,HDL-C,liver pathology,and serum metabolites in rats with non-alcoholic fatty liver disease(NAFLD) induced by high-fat diet(HFD).Methods:50 SD rats were randomly divided into the normal group,the model group,the ST40 group,the ST40 plus ST36 group and the ST40 plus SP6 group,with 10 rats in each group.NAFLD model was established by feeding with HFD in rats.The normal group and the model group received no intervention.The catgut embedding therapy was applied to ST40,ST40 plus ST36 and ST40 plus SP6 respectively in corresponding groups,and the catgut embedding was once a week for a total of 4 weeks during establishing NAFLD model.The serum levels of TC,TG,HDL-C and LDL-C were detected by an automatic biochemical analyzer,and the liver pathological changes of the rats were observed by HE staining and oil red staining.The changes of serum metabolites in rats were analyzed by non-target metabonomics.Results:Compared with those in the normal group,the serum levels of TC,TG and LDL-C were increased(P<0.01),the serum level of HDL-C was decreased(P<0.01),and hepatic fat droplet accumulated in the model group.Compared with those in the model group,the serum levels of TC,TG and LDL-C were decreased in the ST40 group and the ST40 plus SP6 group(P<0.01);however,the serum levels of TC,TG and LDL-C were not decreased significantly in the ST40 plus ST36 group(P>0.05).The three catgut embedding groups had a certain degree of improvement in hepatic fat accumulation,the improvement was most significant in the ST40 plus SP6 group followed by the ST40 group,and the ST40 plus ST36 group showed least improvement.The ST40 plus SP6 group and the ST40 group showed more significant impacts on the choline metabolic pathway(P<0.01),whereas the ST40 plus ST36 group had no significant effects on this pathway(P>0.05).Conclusion:Catgut embedding at ST40 and ST40 plus SP6 can improve the lipid levels of NAFLD rats,whereas ST40 plus St36 can not significantly improve the lipid level of NAFLD rats,which suggest that the combination of ST40 and SP6 has a synergistic effect on NAFLD rats,and the combination of ST40 and ST36 may have a certain antagonistic effect.Metabolomics reveals that the choline metabolic pathway may be the underlying mechanism for its synergistic and antagonistic effects.