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Objective@#To analyze the composition and richness of intestinal microflora in children with non-alcoholic fatty liver disease (NAFLD) and the role of which in pathogenesis of NAFLD.@*Methods@#This was a prospective case-control study. From November 2015 to June 2017, 19 children diagnosed with NAFLD according to the 2010 edition of diagnostic criteria were enrolled voluntarily in the Second and First Affiliated Hospitals of Zhejiang Chinese Medicine University. Twenty-two healthy children were enrolled in the control group. Among the patients, 10 were males and 9 were females, at the mean age of (11.0±1.0) years; 10 males and 12 females in the control group, at the mean age of (9.0±1.2) years. The body mass index (BMI) and waist circumference were recorded, and the fasting blood glucose, total cholesterol, triglyceride, high-density lipoprotein and low-density lipoprotein were detected. Feces were collected and the fecal microorganisms were extracted with magnetic beads methods; the composition and the richness of intestinal microflora in the two groups were detected with 16S rDNA high throughput sequencing technology. The KO differential gene expression and KEGG signal pathway enrichment were analyzed with PICRUST software. The intestinal flora characteristics between the two groups were compared with t test or Mann-Whitney U test and Willcoxon W test.@*Results@#The BMI, waist circumference and triglyceride were higher in NAFLD group than those in the control group (BMI (25.1±2.7) vs. (18.2±1.5)kg/m2, t=9.912, P=0.000; waist circumference (88.6±6.6) vs. (71.5±6.3) cm, t=8.520, P=0.000; triglyceride (0.9±0.4) vs.(0.7±0.3)mmol/L, t=2.060, P=0.046). The abundance and diversity index of intestinal microflora were lower in the NAFLD group (Shannon index 3.99 (3.13, 4.54) vs. 4.63 (4.21, 4.81), Z=-2.065, P=0.039; Simpson index 0.85 (0.70, 0.89) vs. 0.90 (0.88, 0.93), Z=-2.431, P=0.015; ACE index 235.76 (205.26, 361.94) vs. 326.96 (275.34, 368.65), Z=-2.092, P=0.036). At the level of phylum, the proportion of Actinomycetes was lower and the proportion of Thermus was higher in NAFLD group (Actinobacteri 29.807 (14.723, 62.080) ×10-3 vs. 63.212 (46.133, 172.071) ×10-3, Z=-2.667, P=0.008; Thermus 0.033 (0.000, 0.226) ×10-3 vs. 0.000 (0.000, 0.031) ×10-3, Z=-2.729, P=0.006) . At the level of genus, the proportion of Bacteroides and Bifidobacterium in the NAFLD group were significantly lower (Bacteroides 78.757 (11.430, 151.621) ×10-3 vs. 356.821 (161.049, 403.037) ×10-3, Z=-2.771, P=0.006; Bifidobacterium 19.680 (6.181, 53.944) ×10-3 vs. 54.721 (31.911, 146.410) ×10-3, Z=-2.458, P=0.014); the proportion of Prevotella in NAFLD group was significantly higher (3.089 (0.165, 63.502) ×10-3 vs. 0.432 (0.029, 2.257) ×10-3, Z=-2.112, P=0.035). Based on the KEGG database, 78 differentially expressed genes and 26 differential metabolic pathways were found, among which the function genes of K01470, K01961 and K07258 were concentrated in the pathways of arginine and proline metabolism, fatty acid synthesis, and polysaccharides biosynthesis and metabolism. Besides, these three function genes were related to Bacteroides, Prevotella, Bifidobacterium and Ruminococcus.@*Conclusion@#NAFLD children have intestinal flora disturbances in both diversity and abundance, which may alter lipid metabolic pathways through differential gene expressions, contributing to the pathogenesis of NAFLD.
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Objective To explore the effect of high fat diet on serum biochemical parameters and histopathology of main organs in Mongolian gerbils.Methods Forty-eight healthy adult male Mongolian gerbils were randomly and equally divided into model and normal groups.The gerbils in the model group were fed with high fat diet while the normal group with standard diet.Eight gerbils in each group were killed at the end of 4th,8th and 16th week,respectively,and the body weight, serum levels of Glu, TG, CHOL, HDL-C, LDL-C, UA, CREA, BUN, TBil, TP, ALB, ALT, AST and AMS were determined.The histopathological changes of main organs were observed.Results Compared with the normal group,the blood lipid of the model gerbils was significantly increased, the liver function was impaired, the blood uric acid level was higher, and the blood glucose was decreased at the end of 16th week.The AMS was increased at the end of 16th week,but the renal function showed no significant changes.The liver tissue of the model group gradually showed steatosis, inflammation, fibrosis and cirrhosis, accompanied by splenomegalia. The lung tissue and myocardium showed fatty degeneration and obvious damages in the later period,the pancreatic islets were enlarged and the amount of endocrine cells was increased,and the small intestine and kidney didn’ t show any distinct changes.Conclusions A gerbil models of hyperlipidemia and nonalcoholic steatohepatitis and cirrhosis can be well established by high fat diet feeding,and may serve as good models for research of hyperlipidemia-related hyperuricemia, and lung and myocardial damages.
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<p><b>OBJECTIVE</b>To study the expression of uncoupling protein 2 (UCP2) in liver of rats with nonalcoholic steatohepatitis (NASH) induced by fat-rich diet, and the effect of total flavones of hawthorn leafon (TFHL) on UCP2.</p><p><b>METHOD</b>The NASH model of rat was induced by 12 weeks of fat-rich diet. Subsequently the rats were administrated with TFHL in accordance with 250, 125 mg x kg(-1) x d(-1) and the Essentiale N with 195.4 mg x kg(-1) x d(-1). The change of liver pathological. The levels of serum ALT and AST, the content of TG, CHOL, MDA and T-AOC activity of liver and were evaluated. The UCP2mRNA expression in liver was detected with RT-PCR, and the contents of UCP2 were examined with ELISA.</p><p><b>RESULT</b>There are severe steatosis, inflammatory cellular infiltration in the liver of the NASH models. The levels of serum ALT, AST and the contents of TG, CHOL, MDA and UCP2 in the model group were higher than those of in the normal groop. The expression of UCP2mRNA was obviously enhanced and the activity of T-AOC decreased. The expression of UCP2 mRNA of rats was positively correlation with the contents of MDA, TNF-alpha. The inflammation activity in rat liver, the contents of MDA and UCP2, the expression of UCP2 mRNA in the administrated groups were obviously lower than those in the model group, while the activity of T-AOC was higher than that of model.</p><p><b>CONCLUSION</b>TFHL may alleviate liver injury by means of the suppression of Oxidative stress/lipid peroxidation reaction and the overexpression of UCP2 in liver, which could prevent the further development of NASH.</p>
Subject(s)
Animals , Male , Rats , Crataegus , Chemistry , Enzyme-Linked Immunosorbent Assay , Fatty Liver , Drug Therapy , Metabolism , Flavones , Chemistry , Therapeutic Uses , Gene Expression , Ion Channels , Genetics , Liver , Metabolism , Mitochondrial Proteins , Genetics , Plant Leaves , Chemistry , Polymerase Chain Reaction , Rats, Sprague-Dawley , Uncoupling Protein 2ABSTRACT
Objective To study the change of oxidative stress in the formation of NASH in rats induced by high-fat diet,and observe the preventive and therapeutic effect of TFHL on NASH and explore its mechnism.Methods 110 rats were randomly divided into the normal group(40 rats),the model group(40 rats),the A.Nattermam group(10 rats) and high dosage and low dosage TFHL(10rats each).The normal rats were fed with the standard feedstuff the other rats fed with the high-fat diet.At the time of model-making,the rats in the A.Nattermam were fed the A.Nattermam suspension by daily gavage by 195.4mg/(kg?d),the rats in the high dosage and low dosage TFHL group were fed TFHL by daily gavage in 250、125mg/(kg?d),the rats in the normal and model groups were fed isometrical distilled water by daily gavage.At the ends of 2nd,4th,8th weeks of the experiment,10 rats in the nomal and model were killed respectively,at the end of experiment,all the rest were killed.pathological changes of liver tissues were observed,the levels of serum ALT,AST,the amount of TG,CHOL of livers tissue,the amount of MDA of serum and liver,the activity of OD、GSH-PX、T-AOC were measured.Results 1.the rats were fed with high-fat diet from 2 to 12w,the fatty degeneration of liver tissues were gradually serious,the amounts of TG,CHOL of livers were gradually increased.at 12th w the inflammatory cells in livers infiltrated obviously,the serum ALT,AST were obviously increased,it apeared a progressive process of NASH.2.from 4th w,after feeding with high-fat diet,the amounts of MDA of serum and liver were gradually decreased,the activity of SOD were gradually increased,the amount of T-AOC were gradually decreased as well and,increase or decrease with the development of inflammation.3.the inflammation degree of liver,the amount of liver CHOL and the serum ALT,AST in FMCL groups were lower than those in the corresponding model group;the amounts of MDA of serum and liver were decreased,the activity of SOD,GSH-PX and T-AOC were inceased than those in the corresponding model group,their oxidationresisting activity was better than those in the A.Nattermam group.Conclusions oxidation stress/Lipid Peroxidation plays an important role in the occurrence and development of NASH,TFHL can effectively treat and prevent the development of NASH,obviously ease oxidation stress,decrease Lipid Peroxidation of NASH,markedly enhance oxidationresisting abilily of body,which is the important mechanism of prevention and treatment for NASH.
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Objective To investigate the effect of LF on blood biochemistry,cytokines in rats with NASH in order to approach possible mechanism of LF against NASH.Methods 50 male SD rats were randomly divided into the normal control group(10 rats),the model group(10 rats),the Essentinale group(10 rats)、the LF high dose group(10 rats)and the LF low dose group(10 rats).Except for the normal control group which was raised by standard animal feeds,others were all raised by high fat feeds to make NASH model and administered by Essentinale,high dose LF and low dose LF for 12 weeks.Hepatic lipid contents,plasma TG,CHO,HDL-C,LDL-C,ALT,AST and TNF-?,FFA,Leptin concentrations were evaluated,as were histopathological changes in the liver.Results In this NASH model,the liver FZ(liver weight /body weight),plasma CHO,LDL-C,ALT,AST and TNF-?,FFA,Leptin concentrations were higher than that in normal control group,but the levels of serum HDL-C and TG were reduced.In addition,the histopathological showed increased fatty droplets within hepatocytes and liver inflammation.The levels of serum ALT,AST,HDL-C,TNF-?,Leptin,FFA and hepatic CHO,TG contents in the LF high dose group and LF low dose group were reduced compared with that in the model group,while the levels of serum TG,HDL-C were enhanced remarkably.In addition,liver inflammation was improved evidently.Conclusions Treatment with LF might improve non-alcholic steatohepatitis,because LF inhibits the elevation in the level of plasma cytokine.
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Objective:To observe the expression of cytochrome P4502E1 (CYP2E1) in the liver tissue of rats with non-alcoholic steatosis hepatitis (NASH) induced by high-fat diet, and effect of total flavonoids of Chinese hawthorn leaf(TFHL). Methods: The model rats of NASH were induced by 12 weeks' high-fat diet, and treated with TFHL and Essentiale. The pathological changes of rats liver were observed, the amounts of liver MDA, and the activities of SOD, GSH-Px were detected, the mRNA and protein expressions of CYP2E1 were detected with RT-PCR and immunochemical method. Results: ①there were serious steatosis, different degree of infiltration of inflammatory cells and necrosis focus in the livers of model rats, the amounts of liver MDA was higher than that of the normal group, the acttivities of SOD, GSH-Px in the livers were lower, the expressions of mRNA and protein were strengthened. ②the expression of CYP2E1 of livers has a positive correlation with the amount of MDA, but negative correlation with the activity of SOD and GSH-Px. ③the inflammation of liver, the amounts of liver MDA level, the mRNA and protein expressions of CYP2E1,the activity of SOD and GSH-Px in rats treated with high or low dose TFHL were obviously lower than the model rats, but the activities of SOD and GSH-Px were higher than the model rats.Conclussion: CYP2E1 takes part in the onset and development of NASH induced by high-fat diat, TFHL can restrain the activity of CYP2E1 and the lipid peroxidization, enhance the ability of antioxygenation of liver cells, and prevent the further development of NASH.
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Objective:To study the effect of NF-?B and its inhibitor in rats liver during the development of non-alcoholic steato-hepatitis and observe the effect of total flavonoids of Chinese hawthorn leaf on expressions of NF-?B and its inhibitor,and explore the prevention and treatment mechanism of non-alcoholic steato-hepatitis. Methods:Non-alcoholic steato-hepatitis SD rats model were established by administering a high-fat diet for 12 weeks and were treated with total flavonoids of Chinese hawthorn leaf at dosages of 250mg/kg body weight /d,125 mg/kg body weight /d and with Essentiale at a dosage of 195.4mg /kg body weight /d. Pathological changes of liver tissues were observed with HE,the serum MDA,SOD and TNF-? were measured,the protein and mRNA expressions of NF-?B and I?B in rats liver were detected with RT-PCR and immunohistochemical method. Results:There were severe steatosis inflammatory cell infiltration and necrotic foci in non-alcoholic steato-hepatitis rat. Compared with the control group,contents of the serum MDA and TNF-? in non-alcoholic steato-hepatitis group obviously increased,the activity of SOD decreased,the mRNA and protein expressions of NF-?B P65 and I?B? in liver obviously strengthened. There was a positive relativity of the protein expression of NF-?B p65,I?B? with the contents of serum TNF-?,MDA,but a negative relativity with the activity of serum SOD. Conclusion:Total flavonoids of Chinese hawthorn leaf can obviously lower lipid peroxidation in rats,lower the harm of cytokine to hepatocytes,and regulate the expression of protein and mRNA of NF-?B and I?B. It maybe the important mechanism of total flavonoids of Chinese hawthorn leaf for preventing and treating the development of non-alcoholic steato-hepatitis.
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Objective:To study the effect of compound Biejiaruangan tablets on the expression of liver AngiotensionII and its receptor in rats with hepatic fibrosis,and to explore the mechanism of its anti-hepatic fibrosis.Methods: Thirty male SD rats were randomly divided into three groups as followings: the normal group,the model group and the compound Biejiaruangan tablets group,all rats in the two latter groups were given subcutaneous injection of 40%carbon tetrachloride(twice every week for 6 weeks),the rats in the compound Biejiaruangan tablets group were given 1g?kg-1?d-1 of the compound Biejiaruangan tablets by daily gavage,the rats in normal control group and the model group were given distilled water according to the same volume,the histopathological changes in liver were observed through HE and Massion staining.The serum ALT,AST and ALP were evaluated by antomatic biochemistry analysator,the serum levels of hyaluronic acid (HA),Laminin(LN) and plasm AngiotensionII(Ang-Ⅱ) were determined by radio immunoassay.The expressions of AngⅡ,AT1R mRNA were examined by RT-PCR.Results:The severity of inflammation scoring,hepatic fibrosis scoring,the amounts of serum ALT,AST,ALP,HA,LN and plasma AngⅡ in the liver tissues of hepatic fibrosis model group were higher than those in the normal group(P