茵陈蒿汤调控脂肪代谢改善代谢相关脂肪性肝病的效应组分及其机制

Mechanism and components of Yinchenhao Decoction in improving metabolic associated fatty liver disease by regulating fat metabolism

目的:探索茵陈蒿汤调控代谢相关脂肪性肝病(MAFLD)的效应组分及其机制。方法:64只雄性C57BL/6J小鼠随机分为8组,除正常组,其余各组采用高脂饲料喂养12周诱导MAFLD小鼠模型。于第9周首日,正常组和模型组给予去离子水灌胃,其余各组分别给予茵陈蒿汤、绿原酸、大黄多糖、没食子酸、栀子苷及组分复方灌胃,第12周末收集血清和肝组织样本。检测血清肝功能、血脂及肝组织甘油三酯(TG)变化;HE、油红O染色观察肝脏组织病理学变化;UPLC-TQ-MS定量检测血清、肝组织脂肪酸代谢物谱;蛋白免疫印迹法检测肝组织脂肪代谢相关蛋白及核激素受体蛋白表达。结果:模型组肝组织病理,血清ALT、AST、ALP活性及TC、TG、LDL-c、FFA、HDL-c、肝组织TG含量,血清及肝组织乙酸、棕榈油酸浓度,肝组织脂肪代谢相关因子SREBP-1c、ChREBP、ACC1、FAS、CD36、SCD1及核激素受体FXR、LXRα蛋白表达均发生显著改变(P<0.01)。茵陈蒿汤组及各组分均能不同程度地显著改善MAFLD小鼠相关指标变化(P<0.05,P<0.01),其中绿原酸组、大黄多糖组改善血清及肝组织TG、肝组织ACC1及核受体LXRα蛋白表达效果显著优于茵陈蒿汤组、组分复方组及没食子酸组(P<0.05,P<0.01),栀子苷组对血清脂肪酸代谢、肝组织CD36、SCD-1蛋白表达调控作用显著优于其它组分组(P<0.05,P<0.01),大黄多糖组、栀子苷组显著降低小鼠肝组织ChREBP蛋白表达(P<0.05,P<0.01),而没食子酸组、组分复方组、茵陈蒿汤组进一步显著升高模型小鼠肝组织ChREBP蛋白表达(P<0.01)。结论:绿原酸、大黄多糖、栀子苷是茵陈蒿汤治疗MAFLD的效应组分,其机制在于绿原酸和大黄多糖调控FXR/LXRα信号通路,改善脂肪生成代谢,减少新生脂肪形成;栀子苷通过调控脂肪转运及氧化代谢,减少肝细胞脂肪蓄积。

Objective:To explore the effect mechanism of Yinchenhao Decoction and its components in improving metabolic associated fatty liver disease(MAFLD).Methods:Sixty-four male C57 BL/6 J mice were randomly divided into 8 groups.The mice without any treatment was used as control.MAFLD model was induced by high-fat diet for 12 weeks.The other groups were given intragastrically with Yinchenhao Decoction,chlorogenic acid,rhubarb polysaccharide,gallic acid,geniposicle and component compounds,respectively for 4 weeks from week 9,while the control group and the model group were given ddH2 O.At the end of the 12 th week,mice were sacrificed and serum and liver tissue samples were collected.The serum liver function,blood lipids and liver tissue TG levels were measured.The pathological changes were observed by HE and Oil Red O staining.Fatty acid levels in serum and liver tissue were quantitatively measured by UPLC-TQ-MS.The protein expression of fat metabolism related factors and nuclear hormone receptors were detected by Western Blot.Results:Liver pathological changes,serum levels of ALT,AST,ALP activity,TC,TG,LDL-c,FFA,HDL-c,liver TG,serum and liver concentrations of acetic acid and palmitoleic acid and protein expression of fat metabolism related factors SREBP-1 c,ChREBP,ACC1,FAS,CD36,SCD1,as well as the expressions of nuclear hormone receptor FXR,LXRαprotein were dramatically altered in the model group(P<0.01).Compared with that of the model group,all the treatment could improve the changes of related indicators in MAFLD mice(P<0.05,P<0.01).However,chlorogenic acid and rhubarb polysaccharide treatment decreased the serum and liver TG and the protein expressions of ACC1 and LXRαmore efficiently than Yinchenhao decoction,component compound,and gallic acid intervention(P<0.05,P<0.01).Geniposide treatment could normalize the changes of fatty acid metabolism,and protein expressions of CD36 and SCD1 than other components treatment(P<0.05,P<0.01).In addition,rhubarb polysaccharides and geniposide reduced the expression of ChREBP(P<0.05,P<0.01),while gallic acid,component compound,and Yinchenhao Decoction further increased ChREBP protein expression of model mice(P<0.01).Conclusion:Chlorogenic acid,rhubarb polysaccharides and geniposide were the active substances of Yinchenhao Decoction in the treatment of MAFLD,which suggested by the observations that chlorogenic acid and rhubarb polysaccharides regulated lipogenesis to reduce the formation of new fat by FXR/LXRαsignaling pathway;geniposide reduced hepatocytes fat accumulation by regulating fat transport and oxidative metabolism.