Study On The Mechanism Of Shenlingbaizhu Powder In Preventing And Treating Nonalcoholic Fatty Liver Based On Intestinal Microflora And Short-Chain Fatty Acids

Objective:Shenlingbaizhu Powder is regarded as a classic clinical prescription for spleen strengthening.In this study,the changes in serum biochemical indicators,liver pathology,intestinal flora and short-chain fatty acids in model mice given Shenlingbaizhu Power were observed to preliminarily explore the biological basis of the prescription for the treatment of NAFLD.Furthermore,the study provides an experimental basis for spleen therapy based on clinical treatments.Methods:1.Model Reproduction: A high-fat diet had been given on the mice in the experimental group until the end of the experiment,while the control group were given a normal diet.2.Drug Intervention:Start drug intervention on the end of the 4th week.Three drug intervention groups of low,medium,and high-dose were given Shenlingbaizhu Powder for28 consecutive days.Another positive drug group was given bifidobacteria,and one normal group and one model group were given the same amount of normal saline.All the drugs were given through the method of gavage3.Indicator Detection:(1)Serum biochemical indicators: mice serum liver functions(AST,ALT),blood lipids(TC,TG,LDL-C,HDL-C),total bilirubin(TBIL).(2)Pathological indicators: the pathological changes of mice liver were observed through HE and Oil Red O staining methods.(3)Intestinal-micro-ecology-related indicators: the high-throughput gene sequencing was used to detect the abundance and diversity of mice intestinal flora.The gas-mass spectrometry coupled with chromatography was used to detect the content of short-chain fatty acids in the intestinal contents.Results:1.Changes in Serum Biochemical Indicators(1)The serum biochemical indexes ALT,AST,TC,TG,LDL-C and HDL-C of the mice in the model group and the drug intervention groups increased significantly(P<0.01)compared to the normal group.The index TBIL in the model group increased significantly(P<0.01),while there was no significant difference in other groups(P>0.05).(2)The serum biochemical indexes ALT,AST,TC,TG,LDL-C,HDL-C and TBIL of the low-dose and medium-dose drug intervention groups showed a significant downward trend(P<0.01)compared to the model group.The indexes AST,TC,TG,LDL-C,HDL-C,TBIL,and Glu in the high-dose drug intervention group showed a significant downward trend(P<0.01),while no significant difference was observed of index ALT(P>0.05).Indexes AST,TC,TG,LDL-C,HDL-C and TBIL,significantly decreased in the bifidus group(P<0.01),there is also a decrease in index ALT(P<0.05).2.Changes in Pathological Indicators(1)Fatty vacuoles,disordered liver cord arrangement,partly-ruptured hepatocytes were developed in the mice of the model group.Also,part of the color-darkened area after the hepatocyte nucleus staining showed inflammatory factor infiltration.(2)Compared with the model group,the liver cell structure,hepatic cord arrangement and liver lobule structure in the low-dose intervention group had no obvious abnormalities.There are small scattered fatty vacuoles with clear liver cords in the medium-dose group.The small fatty vacuoles can be developed in the high-dose intervention group,but the vacuoles seem to be smaller and less than the vacuoles in the model group.In the bifidus group,a small amount of fatty vacuoles can be developed,but the morphology of liver cells was normal,and the liver cords were arranged regularly.3.Changes in intestinal flora(1)The changes in the phyla level abundance: Compared to the normal group,the relative abundance level of two phyla of the mice intestinal contents in the model group increased,while the relative abundance level of other eight phyla decreased.The relative abundance level of two phyla of the intestinal contents of mice in the drug intervention groups increased compared to the model group,while the relative abundance level of two phyla decreased.In the bifidus group,the relative abundance level of three phyla increased and the level of four phyla decreased.(2)The changes in the level of genus abundance: Compared to the normal group,the relative abundance level of 16 bacterial genera mainly composed of Lachnospiraceae NK4A136 group(8.70%),Dubosiella(2.74%)and Helicobacter(5.08%)increased in the mice intestinal flora of the model group.The relative abundance level of four genera mainly composed by Lactobacillus(28.66%)decreased.Compared to the model group,the relative abundance level of five bacterial genera mainly composed by Lachnospiraceae NK4A136 group,Dubosiella and Lactobacillus in the drug intervention group increased,while the relative abundance level of nine genera mainly composed by Helicobacter and Lachnospiraceae UCG-008 decreased.Finally,the relative abundance level of nine bacterial genera mainly composed by Dubosiella and Lactobacillus in the bifidus group increased,and the level of 11 genera mainly composed by NK4A136 group and Helicobacter decreased.4.Changes in SCFA contentCompared with the normal group,the contents of Propionic acid and Butyric acid in the model group were significantly increased(P<0.01).Compared with the model group,propionic acid in the low,medium and high reference groups and bifidum groups tended to decrease(P>0.05),acetic acid and butyric acid had no obvious trend.Conclusion:1.Shenlingbaizhu Powder may improve the liver fatty degeneration of the mice of the NAFLD model.2.Shenlingbaizhu Powder is based on the treatment method of "curing liver and strengthening spleen" to strengthen spleen and treat NAFLD.Its action mechanism may be related to the regulation of intestinal microecology,which is specifically related to the increase of Bifidobacteria and Lactobacillus,the decrease of Helicobacteria,and the change of short-chain fatty acid content.