Proteomics analysis on hypolipidemic mechanisms of total phenylpropanoid glycosides from Ligustrum robustum(Roxb.) Blume in hamsters fed a high fat diet

OBJECTIVE To explore the hypolipidemic mechanisms of the total phenylpropanoid glycosides fromLigustrum robustum(Roxb.) Blume(LRTPG) in hamsters using proteomics technique.METHODS The hamsters were fed with a high fat diet to induce hyperlipidemia.Then LRTPG of high(1.2 g·kg^(-1)),medium(0.6 g·kg^(-1)) and low(0.3 g·kg^(-1)) doses were administrated daily for 4 weeks.Then the concentrations of plasma and hepatic lipids were determined using enzymic methods.The total protein was extracted from livers of the model group and the group treated with the high dose of LRTPG for label-free quantitative proteomics.RESULTS LRTPG significantly reduced the concentrations of plasma and hepatic lipids in hamsters fed a high fat diet.The proteomics data showed that a total of 2231 proteins were identified,and 549 proteins were found to be differentially expressed between the model group and the group treated with LRTPG.Among the 549 proteins,93 proteins were up-regulated and 59 proteins were down-regulated,and 397 proteins were absent or not.And some of these proteins were much related to the lipid metabolism.Further,gene ontology(GO) analysis indicated metabolic process,transport,oxidation-reduction process,phosphorylation,signal transduction,lipid metabolic process were the main biological processes that those differentially expressed proteins participated.KEGG pathway analysis showed that those proteins were involved in several metabolic pathways including oxidative phosphorylation,non-alcoholic fatty liver disease(NAFLD),PI3K-Akt signaling pathway,cAMP signaling pathway,cGMP-PKG signaling pathway.CONCLUSION The proteomics study could provide valuable clues to help us to understand the hypolipidemic mechanisms of LRTPG much better.

基于肠道菌群探讨粗壮女贞总苷对高脂血症金黄仓鼠的降脂作用机制

目的评价粗壮女贞总苷(total phenylpropanoid glycosides extracted from Ligustrum robustum(Roxb.)Blume,LRTPG)对高脂血症金黄仓鼠的降脂作用并探讨其对肠道菌群的调节作用。方法60只仓鼠随机分为对照组、模型组、阳性药组、LRTPG-L组、LRTPG-M组、LRTPG-H组。高脂饲料诱导造模成功后连续给药4周,进行血脂和肝脂含量检测,并采集对照组、模型组和LRTPG-H组仓鼠结直肠部位成形粪便进行16S rDNA测序。结果LRTPG对高脂血症仓鼠具有降低血清TG、TC、LDL-C水平和肝脏TG、TC含量的作用。肠道菌群测序结果显示,与模型组比较,LRTPG在门水平降低厚壁菌门相对丰度,增加拟杆菌门和疣微菌门相对丰度(P<0.01);科水平上明显升高克里斯滕森菌科、消化球菌科、疣微菌科相对丰度(P<0.05或P<0.01);属水平上明显升高颤螺菌属、颤杆菌克属、解黄酮菌属和阿克曼氏菌属的相对丰度(P<0.05或P<0.01)。这些菌群的变化均有益于LRTPG降脂作用的发挥。结论LRTPG可能通过改善高脂饮食导致的金黄仓鼠肠道菌群紊乱发挥降脂功效。

粗壮女贞总苷降脂作用及其基于AMPK通路的降脂作用机制研究

目的探讨粗壮女贞总苷(total phenylpropanoid glycoside from CN,CNTG)降脂作用及其基于AMPK通路的降脂作用机制。方法 Syrian金黄地鼠随机分为正常组、高脂模型组、阳性药非诺贝特组及CNTG高、中、低剂量组。除正常组外,其余各组地鼠应用高脂饲料诱导建立混合型高脂血症模型。造模1周后取血,检测血清TC、TG、LDL-C、HDL-C含量,确定模型形成。之后连续给药4周,在给药2周、3周及4周后分别取血,检测血清中TC、TG、LDL-C和HDL-C的含量。在给药4周后剖取肝脏,进行肝脏TG、TC含量测定和肝脏病理学检测。并应用实时定量PCR检测CNTG对肝脏中AMPK、CD36、CPT1、ACC的mRNA表达的影响;用Western blot法检测AMPK和其上游调控蛋白LKB1的磷酸化蛋白和总蛋白水平,并检测HMG-Co A还原酶的蛋白表达量。结果给药2周、3周、4周后,与模型组相比,CNTG高剂量组血清中的TC、TG、LDL-C含量均明显降低(P<0.05,P<0.01);肝脏中TC、TG含量明显降低(P<0.01);并可明显减轻肝细胞病变。实时定量PCR结果显示,与模型组相比,CNTG可以明显升高AMPK的相对表达量(P<0.01),但CD36、ACC和CPT1的相对表达量无明显变化。Western blot结果显示,与模型组相比,CNTG高剂量组肝脏中的LKB1、phospho-LKB1及phospho-AMPKα的相对表达量均提高(P<0.05),差异具有显著性。结论 CNTG能有效降低高脂血症金黄地鼠的血脂和肝脂水平,其机制可能是CNTG促进肝脏中LKB1磷酸化以激活AMPK,从而调控机体脂质代谢。

基于蛋白质组学探讨粗壮女贞总苷对高脂血症金黄地鼠的调脂作用机制

目的 应用非标记定量(label-free)蛋白质组学技术,探究粗壮女贞总苷(LRTPG)的调脂机制。方法 从高脂血症模型组和LRTPG给药组金黄地鼠肝脏中提取总蛋白,进行label-free蛋白质组学研究。结果 蛋白质组学研究共鉴定出2 231个蛋白质,模型组与LRTPG组共549个差异表达蛋白,其中93种蛋白上调,59种蛋白下调,397种蛋白只在模型组或者给药组中有定量值。GO分析表明,这些差异表达蛋白主要参与代谢、转运、氧化还原、磷酸化、信号转导、脂质代谢等生物学过程。KEGG通路分析表明,这些蛋白集中于氧化磷酸化、非酒精性脂肪肝病、PI3K-Akt、cAMP、cGMP-PKG等多条信号通路。一些差异蛋白与脂质代谢密切相关,包括CD36、PK、HSS、GCK、ApoA I、Acly、FABP5等。结论 LRTPG调脂作用可能与CD36、PK、HSS、GCK、ApoA I、Acly、FABP5有关。