Huatan Huoxue Tongluo Recipe(化痰活血通络方)Regulates Intestinal Microflora and Inhibits LPS Mediated Inflammatory Pathway to Improve Vascular Injury in Atherosclerosis Mice
Objective To explore the mechanism of Huatan Huoxue Tongluo Recipe(化痰活血通络方)in treating atheroscle-rosis(AS)from intestinal flora and provide a basis for the clinical application of Huatan Huoxue Tongluo Recipe.Method Twenty-five male ApoE-/-mice were fed high fat diet for 12 weeks to prepare AS models.The model mice were divided into model group(MX),whole recipe group(QF),dredging collateral group(TL),resolving phlegm group(HT)and atorvastatin group(XY)with 5 mice each.In addition,5 C57BL/6J male mice were set as the normal group(ZC),and each group was treated with the corresponding drugs by gavage for 12 weeks.The gut microbiota was detected by using 16S rRNA amplicon sequencing.The pathological morphology of mouse aorta and intestine was observed by using hematoxylin eosin(HE)staining method.Enzyme-linked immunosorbent assay(ELISA)was used to detect relevant indicators in mouse serum.Western blot was used to detect Toll like receptor 4(TLR4),NOD like receptor heat protein domain related protein 3(NLRP3),nuclear factors-κB(NF-κB)and interleukin-6(IL-6)protein contents.Result Compared with those in the blank group,the species richness and diversity in the MX group decreased significantly,and the proportion of Firmicutes to Bacteroidetes(F/B)increased significantly.Lipopo-lysaccharides(LPS),triglycerides(TG),total cholesterol(TC)and low-density lipoprotein cholesterol(LDL-C)in the serum as well as TLR4,NLRP3 and NF-κB in the aorta were observed in the MX group.The content of IL-6 protein increased signif-icantly(P＜0.01),the content of high-density lipoprotein cholesterol(HDL-C)decreased significantly(P＜0.01),the area of aortic plaque in mice increased significantly(P＜0.01),accompanied by a large number of foam cells and inflammatory infiltra-tion,large areas of villus shed and lamina propria exposed in the intestine.Compared with the MX group,the QF group could sig-nificantly increase the species richness and diversity,reduce the proportion of F/B(P＜0.01),and increase the relative abun-dance of beneficial bacteria,reduce the relative abundance of harmful bacteria,including LPS,TG,TC,LDL-C,TLR4,NLRP3 and NF-κB in the QF group and XY group.IL-6 significantly decreased(P＜0.01),HDL-C significantly increased(P＜0.01),and the above pathological changes were significantly alleviated.Conclusion Huatan Huoxue Tongluo Recipe can improve the composition of intestinal microbiota and lipid metabolism and reduce the production of LPS,and it can down regulate the ex-pression of inflammatory factors in the aorta and reduce the accumulation of lipids to alleviate atherosclerosis in ApoE-/-mice through TLR4/NF-κB/NLRP3 signaling pathway.
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