基于肠道菌群研究全缘叶绿绒蒿提取部位干预非酒精性脂肪性肝病的作用

Study on the effect of the extraction sites of Meconopsis integrifolia (Maxim.)French in nonalcoholic fatty liver disease based on intestinal microbiota

关键词:  全缘叶绿绒蒿  提取部位  非酒精性脂肪肝  肠道菌群

Key words:  Meconopsis integrifolia (Maxim.)French  extraction site  non-alcoholic fatty liver disease  gut microbiome

摘要:

摘要：目的 以全缘叶绿绒蒿为研究对象，以肠道菌群为目标，通过细胞、动物水平的药理学研究，探讨全缘叶绿绒蒿提取部位对非酒精性脂肪肝病（Nonalcoholic fatty liver disease，NAFLD）大鼠及其肠道菌群（Gut microbes，GM）构成的影响。方法 全缘叶绿绒蒿萃取后得到乙酸乙酯部位、正丁醇部位，通过棕榈酸（Palmitic acid，PA）刺激HepG2细胞建立体外脂质沉积模型，在细胞水平上测定给予提取部位后细胞中的脂质的含量、抗氧化能力和炎症因子表达水平。通过高脂饲料（High fat diet, HFD）建立NAFLD大鼠模型，在动物水平上测定提取部位干预后大鼠体重、转氨酶活力、血脂含量、抗氧化能力和炎症因子表达水平，并通过16s高通量测序和GC-MS对大鼠肠道菌群及短链脂肪酸（Short chain fatty acid，SCFA）进行表征。结果 全缘叶绿绒蒿乙酸乙酯、正丁醇部位能降低甘油三酯（Triglyceride，TG）、低密度脂蛋白胆固醇（Low-density lipoprotein cholesterol，LDL-C）、丙二醛（Malondialdehyde，MDA）的含量，升高超氧化物歧化酶（Superoxide dismutase，SOD）、谷胱甘肽（Glutathione，GSH）的水平，降低肿瘤坏死因子-α（Tumor necrosis factor-α，TNF-α）、白介素-6（Interleukin-6，IL-6）、白介素-1β（Interleukin-1β，IL-1β）的水平。同时降低大鼠体重，减少肝组织脂质沉积，降低谷草转氨酶（Aspartate aminotransferase，AST）、谷丙转氨酶（Alanine aminotransferase，ALT）水平，提高高密度脂蛋白（High-density lipoprotein cholesterol，HDL-C）含量。在GM上，全缘叶绿绒蒿干预后可降低Romboutsia、RF39_unclassified丰度，提高Allobaculum、Ruminococcus、Lachnospiraceae_NK4A136_group丰度，同时还能提高SCFA的含量。结论 全缘叶绿绒蒿可能通过调节NAFLD大鼠GM相对丰度进而影响菌群代谢产物SCFA，降低炎症因子表达水平，提高抗氧化能力以达到对NAFLD大鼠的保护作用。

Abstract:

ABSTRACT: OBJECTIVE: To investigate the effects of the extraction sites of Meconopsis integrifolia (Maxim.)French on NAFLD and the composition of the gut microbes (GM) through pharmacological studies at the cellular and animal levels. METHODS: Ethyl acetate site and n-butanol site were obtained from Meconopsis integrifolia (Maxim.)French by extraction. HepG2 cells were stimulated by palmitic acid (PA) to create a cell lipid deposition model, and intracellular lipid content, antioxidant capacity, and inflammatory factor expression levels were measured. The rat model of nonalcoholic fatty liver disease (NAFLD) was established with high fat food (HFD), and the body weight, transaminase activity, blood lipid content, antioxidant capacity and inflammatory factor expression level of the rats were determined, and the gut microbes and short chain fatty acid (SCFA) of the rats were characterized by 16s high-throughput sequencing and GC-MS. RESULTS: The ethyl acetate sites and n-butanol sites of Meconopsis integrifolia (Maxim.)French could reduce the contents of triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), malondialdehyde (MDA), increase the levels of superoxide dismutase (SOD) and glutathione (GSH) and decrease the levels of tumor necrosis factor-α, interleukin-6 and interleukin-1β. At the same time, the body weight of rats, the lipid deposition of liver tissue and the levels of aspartate aminotransferase and alanine aminotransferase were reduced, and the content of high-density lipoprotein cholesterol (HDL-C) was increased. In GM, the abundance of Romboutsia and RF39_unclassified was reduced, the abundance of Allobaculum, Ruminococcus and Lachnospiraceae_NK4A136_group was increased after intervention of Meconopsis integrifolia (Maxim.)French, and the content of SCFAs was also increased. CONCLUSION: Meconopsis integrifolia (Maxim.)French may affect microbiota metabolites, reduce the expression level of inflammatory factors, and improve antioxidant capacity to achieve a protective effect on NAFLD rats by regulating the relative abundance of GM in NAFLD rats.