| Objecive:The effects of alcohol extracts of Mesona chinensis(MC)on lipid levels,liver lipid metabolism,liver pathological changes,serum free fatty acids,serum fatty acylcarnitine and fecal short chain fatty acids changes in rats with hyperlipidemia induced by high-fat diet were evaluated.The 16S DNA high-throughput sequencing method was used to compare the effects of different doses of ethanol extract of MC on the abundance and diversity of gut microbiota in rats fed with high fat diet.The antioxidative activities and the inhibitory effects on α-glucosidase,α-amylase and pancreatic lipase in the different polar parts of the extract of MC were investigated to study the anti-oxidation and inhibition of enzyme in vitro.Methods:1.Therapeutic effect of ethanol extract of MC on hyperlipidemia rats 1.1 The lipid-lowering experiment on rats induced by high-fat diet with the alcohol extract of MC48 SD male rats were selected and normal control group 8.And the remaining 40 are modeled.After successful modeling,they are divided into 5 groups(8 in each group).namely,model group.simvastatin group(8mg·kg-1·d-1),low-,medium-,and high-dose groups of MC extract(The low.medium and high doses of alcohol extract were 2,4,8g·kg-1·d-1,respectively.)The normal control group was fed the basal diet.and the other groups were fed a high-fat diet for 4 weeks.Experimental analysis was performed after the end of the experimental administration.1.2 Quantitative analysis of serum free fatty acids in high-fat rats by alcohol extract of MCThe content of free fatty acids in the serum of each group was determined by gas chromatography-mass spectrometry(GC-MS),and investigated the effect of MC on serum free fatty acids in rats fed with high fat diet.1.3 Quantitative analysis of serum fatty acylcarnitine in high-fat rats by alcohol extract of MCThe content of fatty acylcarnitine in the serum of each group was determined by ultraviolet liquid chromatography-tandem mass spectrometry(UPLC-MS/MS),and the effect of MC on the fatty acylcarnitine in the serum of rats with high fat diet was investigated.1.4 Quantitative analysis of fecal short chain fatty acids in high-fat rats by alcohol extract of MCThe content of short chain fatty acids in the fecal of each group was determined by gas chromatography-mass spectrometry(GC-MS),and investigated the effect of MC on fecal short chain fatty acids in rats fed with high fat diet.1.5 Regulation of the extract of MC on the gut microbiota of high-fat ratsUsing 16S rDNA high-throughput sequencing method to compare the effects of different doses of MC on the abundance and diversity of gut microbiota in rats with high-fat diet.And the abundance and diversity of gut microbiota of rats were analyzed by using theⅢumina sequencing platform.2.Study on antioxidative activity and the inhibition of α-glucosidase,α-amylase and pancreatic lipase of different polar parts extraction of MC.2.1 Antioxidant activity of different polar parts of MCThe DPPH free radicals,ABTS+radical scavenging ability and FRAP value were determined in different polar parts of MC,to investigate the antioxidant capacity of different parts.2.2 Inhibition of α-glucosidase,α-amylase and pancreatic lipase in different polar parts of MCThe inhibition of α-glucosidase,pancreatic lipase and α-amylase by different polar parts was determined by PNPG method,PNPP method and DNS method,respectively.Result:1.Therapeutic effect of ethanol extract of MC on hyperlipidemia rats1.1 The serum levels of TC,TG and LDL-C in the obese rats induced by high-fat diet were significantly higher than those in the normal group,and HDL-C was significantly decreased.The MC had a certain inhibitory effect on obesity caused by high-fat diet.The high.medium and low doses of MC alcohol extract group could significantly reduce the levels of serum TC,TG and LDL-C in rats.HDL-C levels were significantly elevated in the low-dose MC alcohol extract group.The TG content in the liver was significantly reduced in all doses of MC,and the pathological section of the liver further confirmed the liver protection of the MC and reduced liver damage.1.2 The contents of saturated fatty acids such as myristic acid,pentadecanoic acid,palmitic acid,heptadecanoic acid,stearic acid,arachidic acid,lignoceratic acid and triacontanic acid in serum of high-fat model rats was significantly increased.while the contents of unsaturated fatty acid such as palmitoleic acid,trans-9-elaidic acid,cis-9-oleic aicd,linoleic acid,arachidic acid were significantly decreased.After the treatment with the MC extract,the content of saturated fatty acids and unsaturated fatty acids in the serum of the MC administration group tended to be normal.1.3 35 kinds of fatty acylcarnitines were obtained by analysis and screening.The main fatty acylcarnitine was free carnitine.which accounted for 50%of total fatty acylcarnitine.followed by acetylcarnitine,which accounted for 40%of total fatty acylcarnitine.Compared with the NC group,the CO(free carnitine)content in the HFD group was significantly lower,and the PC group and LMC and MMC were significantly higher than the HFD group.In short-chain fatty acylcarnitine,compared with HFD,the content of C2:0(acetylcarnitine),C4:0(butyrylcarnitine)and C5i:0(isovalerate carnitine)in the serum of MMC and HMC group were significantly decreased,and the C5i:0 content of the LMC group was also significantly decreased.In the long-chain fatty acylcarnitine,C18:0(octadecacarnitine)in the serum of the HFD group was significantly increased compared with the NC group,C18:1(octadecyl-enoylcarnitine)Significantly decreased;compared with the HFD group,serum C18:0 was significantly increased in the MMC group,and C18:1 in the MMC and HMC group was significantly increased.There were no significant differences in other fatty acylcarnitine contents.1.4 The results of determination of short-chain fatty acid in rats showed that the content of acetic acid and butyric acid in HFD group was significantly lower than that in NC group,and the contents of propionic acid,isobutyric acid.isovaleric acid and valeric acid were all decreased.The increase in the content of isobutyric acid and isovaleric acid was significant;compared with the HFD group,the acetic acid content was significantly increased after the ethanol extract of the MC,the butyric acid level increased,but there was no significant difference,and the levels of propionic acid,isobutyric acid,isovaleric acid and valeric acid in the MMC and HMC group increased significantly.1.5 The high-fat diet reduced the abundance and diversity of the gut microbiota in rats.After the administration of the MC extract,the number and diversity of the gut microbiota of the rats were increased.The difference of bacterial between the drug-administered group and the normal group was small.It indicated that MC can improve the imbalance of gut microbiota in rats and form a relatively stable gut microbiota structure,Among them,at the level of the phylum,the Firmicutes was significantly decreased,Bacteroidetes was significantly increased;at the level of the class,Clostridia significantly decreased.Bacteroidia significantly increased;at the order level,HMC group Clostridiales significantly decreased,and Bacteroidales significantly increased.At the level of the family.the Lachnospiraceae was significantly decreased,and the Muribaculaceae was significantly increased;at the genus level,the proportion of the Lachnospiraceae was decreased,Muribaculaceae was significantly increased.It was found that there were scarcely Verrucomicrobia and Blautia genus in the normal group,and the abundance was significantly increased after feeding the high-fat diet.2.Antioxidant activity and inhibition of α-glucosidase,α-amy lase and pancreatic lipase in different polar parts of alcohol extract of MC.The results showed that the strongest antioxidant capacity in different polar parts of MC is ethyl acetate,and the weakest is petroleum ether.With the increases of the same concentration,the antioxidant capacity of each part was enhanced.The most inhibitory effect on α-glucosidase is the n-butanol site,the weakest is the water site.The most inhibitory effect on α-amylase is the ethyl acetate site,the weakest is the chloroform site.The strongest inhibitor of lipase is the ethyl acetate site,the weakest being the petroleum ether site and the water site.And as each part increases with the concentration of the sample.the ability of inhibit the enzyme was enhanced.Overall,the ethyl acetate part and the n-butanol part were the most potent.Conclusion:Alcohol extract of MC can significantly reduce the lipid level of hyperlipidemic rats induced by high-fat diet,improve abnormal changes of serum free fatty acids.regulate changes in short-chain fatty acids in the feces,and improve gut microbiota imbalance.And alcohol extract and other polar parts have good antioxidant activity,and have good inhibitory effects on α-glucosidase,α-amylase and pancreatic lipase. |
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