The Mechanism Of Pueraria Regulating Hyperlipidemia Based On Intestinal Bacteria Metabolism

Purpose:In this study,an animal model was used to investigate whether pueraria(pueraria extract and puerarin)could regulate hyperlipidemia by regulating the changes of intestinal flora and short-chain fatty acids,repairing intestinal mucosal barrier,reducing the level of systemic inflammation to further interfer m RNA expression level of genes related to lipid metabolism,thus to preliminarily explore the molecular mechanism of pueraria regulating hyperlipidemia.Methods:The HLP mouse model was successfully replicated by HFD feeding for 10 weeks,and then pueraria was given as intervention for 4 weeks.UHPLC-QTOF-MS non-target metabonomics method was used to detect the active components in pueraria.Serum levels of total cholesterol(TC),low density lipoprotein cholesterol(LDL-C)and high density lipoprotein cholesterol(HDL-C)in mice were determined by the test kit.HE staining and oil red O staining pathological sections were made to observe the degrees of inflammatory cell infiltration and lipid deposition in liver tissues as well as the degrees of inflammatory cell infiltration and intestinal mucosal barrier injury in intestinal tissues.The bacterial 16 S r RNA microbiome diversity sequencing method was used to detect the abundance and diversity of the bacterial community in the intestinal contents of mice.GC-MS high-throughput target metabolomics was used to detect the change of short-chain fatty acids in the intestinal contents of mice.The contents of secretory immunoglobulin A(s Ig A)and serum tumor necrosis factor(TNF-α),lipopolysaccharide(LPS)in intestinal tissues of mice were determined by ELISA.Real-time fluorescent quantitative PCR(Q-PCR)method was used to detect the relative m RNA expression levels of SREBP-1c,Lx R,IL-18,CYP7A1 and etc..Western blot(WB)was used to detect the protein expression levels of ZO-1,Occludin and NLRP3 in the intestinal tissues of mice.The intestinal flora of mice was eliminated with quadruple antibiotics.Results:Compared with model group,serum levels of TC and LDL-C decreased significantly(P<0.05)and HDL-C level increased significantly(P <0.01)after pueraria intervention,and the degrees of vacuoloid changes,lipid deposition and inflammatory cell infiltration in liver tissues were reduced.Compared with model group,pueraria intervention group showed increased abundance and decreased diversity of intestinal flora as well as increased contents of short-chain fatty acids such as acetic acid and propionic acid.Compared with model group,pueraria could significantly reduce the infiltration of inflammatory cells in intestinal tissues and the expression levels of NLRP3,Caspase-1,IL-18 and etc.,increasing the expression levels of ZO-1,Occludin and s Ig A,repairing the intestinal mucosal barrier and reducing the entry of LPS into the blood in HLP model mice.After the intervention of pueraria in mice,the expression of cholesterol absorption related genes in liver was down-regulated and the expression of metabolism-related genes was increased.Antibiotics were used to clear the intestinal flora of mice,and then the HLP mouse model was replicated and intervened with pueraria.The results showed that the blood lipid level,liver inflammatory cell infiltration,vacuole-like changes and lipid droplet deposition were more serious in bacteria-disinfected model group than those in model group.Compared with pueraria intervention group,the effect of pueraria intervention was weakened after clearing the bacteria.Compared with model group,bacteria-disinfected model group showed more serious inflammation expression and intestinal mucosal injury in intestinal tissues,increased entry of LPS into the blood,and increased serum inflammation levels.Compared with pueraria intervention group,the effects of reducing inflammation levels in intestinal tissues and serum as well as repairing intestinal mucosal injury by pueraria were weakened after clearing the bacteria.At the same time,compared with model group,the relative expression level of m RNA of cholesterol synthesis-related genes increased,and the relative expression level of m RNA of decomposition-related genes decreased.Compared with pueraria intervention group,the effect of pueraria intervention was weakened after clearing the bacteria.Conclusion:1.Pueraria can repair the intestinal mucosal barrier in HLP model mice,and reduce the inflammation level in intestinal tissues as well as the entry of endotoxin into the blood to alleviate the body’s chronic metabolic inflammation.2.Pueraria may regulate the expression of liver lipid metabolism genes by intervening intestinal bacteria metabolism.3.Pueraria can regulate the abundance and diversity of intestinal flora,increasing the content of short-chain fatty acids,repairing intestinal mucosal barrier,reducing inflammation of intestinal tissues and entry of endotoxin into the blood to reduce systemic inflammation,and relieving hyperlipidemia by intervening m RNA expression level of liver lipid metabolism gene to reduce blood lipid level.