基于脂质组学研究黄芩-黄连药对的降脂作用

Lipid-lowering effect of drug pair Scutellariae Radix-Coptidis Rhizoma based on lipomics

基于脂质组学研究黄芩-黄连药对干预非酒精性脂肪肝病(nonalcoholic fatty liver disease,NAFLD)大鼠的作用机制。将36只SD大鼠分为空白组、模型组、不同剂量黄芩-黄连组和辛伐他汀组,每组6只。除空白组给予普通饲料外,其余各组给予高脂饲料,饲喂4周后,开始给予药物干预,12周后处理。使用试剂盒检测血清肝功能和血脂指标,通过HE染色与油红O染色法评价肝组织的病理形态。使用LC-MS/MS技术检测大鼠脂质水平变化,采用多元统计分析筛选其中的差异脂质代谢物,并绘制脂质代谢通路。进一步通过Western blot验证脂质相关的蛋白水平变化。结果表明,与空白组比较,模型组大鼠谷丙氨酸氨基转移酶(alanine aminotransferase,ALT)、天门冬氨酸氨基转移酶(aspartate aminotransferase,AST)、总胆固醇(total cholesterol,TC)、甘油三酯(triglyceride,TG)、低密度脂蛋白胆固醇(low-density lipoprotein cholesterol,LDL-c)水平显著升高(P<0.01),γ-谷氨酰基转移酶(gamma-glutamyl transferase,γ-GT)、高密度脂蛋白胆固醇(high-density lipoprotein cholesterol,HDL-c)含量显著降低(P<0.01);黄芩-黄连干预后显著回调。HE染色与油红O染色表明不同剂量的黄芩-黄连均可逆转大鼠肝脏的脂肪性病变,且呈剂量依赖性。脂质组学分析发现模型组与空白组大鼠的脂质代谢存在明显差异,有54个脂质发生了显著改变,主要有甘油脂类、磷脂酰胆碱类、鞘脂类等。给药后,有44个脂质差异物趋于正常水平,这表示不同剂量的黄芩-黄连给药后显著改善了NAFLD大鼠的脂质代谢水平。Western blot结果表明,黄芩-黄连能显著降低甘油三酯合成酶1(TG-synthesis enzyme 1,DGAT1)、脂素1(recombinant lipin 1,LPIN1)、脂肪酸合成酶(fatty acid synthase,FASN)和乙酰辅酶a羧化酶1(acetylCoA carboxylase 1,ACC1)蛋白的水平,升高ACC1的磷酸化水平,这些变化可显著降低TG的合成,且提高其分解速度,提升机体的脂质代谢水平,最终实现降脂的作用。黄芩-黄连能够通过抑制脂肪酸和TG的合成改善NAFLD。

This study investigated the mechanism of action of Scutellariae Radix-Coptidis Rhizoma(SR-CR)in intervening in nonalcoholic fatty liver disease(NAFLD)in rats based on lipidomics.Thirty-six SD rats were divided into a control group,a model group,SR-CR groups of different doses,and a simvastatin group,with six rats in each group.Rats in the control group were fed on a normal diet,while those in the remaining groups were fed on a high-lipid diet.After four weeks of feeding,drug treatment was carried out and rats were sacrificed after 12 weeks.Serum liver function and lipid indexes were detected using kits,and the pathomorphology of liver tissues was evaluated by hematoxylin-eosin(HE)staining and oil red O staining.Changes in lipid levels in rats were detected using the LC-MS technique.Differential lipid metabolites were screened by multivariate statistical analysis,and lipid metabolic pathways were plotted.The changes in lipid-related protein levels were further verified by Western blot.The results showed that compared with the control group,the model group showed increased levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),total cholesterol(TC),triglyceride(TG),and low-density lipoprotein cholesterol(LDL-c)(P<0.01),and decreased levels ofγ-glutamyl transferase(γ-GT)and high-density lipoprotein cholesterol(HDL-c)(P<0.01),which were significantly recovered by the intervention of SR-CR.HE staining and oil red O staining showed that different doses of SR-CR could reverse the steatosis in the rat liver in a dose-dependent manner.After lipidomics analysis,there were significant differences in lipid metabolism between the model group and the control group,with 54 lipids significantly altered,mainly including glycerolipids,phosphatidylcholine,and sphingolipids.After administration,44 differential lipids tended to normal levels,which indicated that SR-CR groups of different doses significantly improved the lipid metabolism level in NAFLD rats.Western blot showed that SR-CR significantly decreased TG-synthesis enzyme 1(DGAT1),recombinant lipin 1(LPIN1),fatty acid synthase(FASN),acetyl-CoA carboxylase 1(ACC1),and increased the phosphorylation level of ACC1.These changes significantly decreased the synthesis of TG and increased the rate of its decomposition,which enhanced the level of lipid metabolism in the body and finally achieved the lipid-lowering effect.SR-CR can improve NAFLD by inhibiting the synthesis of fatty acids and TG.