| Objective: to observe the effect of compound Nutrient Diet on Ob/Ob mouse model of NAFLD and explore the mechanism of its effect on SIRT1 / FOXO1 pathway.Methods: 20 male C57BL/6 mice and 20 Ob / Ob mice of SPF 4~8 weeks old were randomly divided into 4 groups: Normal Control Group,Normal Vegetarian Group,model group and Model Vegetarian Group.Normal Control Group and model control group were fed with normal diet,normal vegetarian group and model vegetarian group were fed with compound nutrient diet,and the food intake and body weight was recorded weekly.Two groups of mice were killed at 1 month and 3 months after intervention.5 Mice in each group were killed by random sampling.Weight,length and body mass index were measured.Separate the liver,weigh the wet weight of the liver,and calculate the liver index.Serum samples for Aspartate Transaminase,Alanine Transaminase,Cholesterol.H&E,oil red O staining and Masson staining were used to observe the pathological changes of Fatty Change,balloon-like change,inflammation and liver fibrosis in left lobe liver.The expression of SIRT1 mRNA was detected by RT-PCR,the protein levels of SIRT1 and AC-FOXO1 by Western Blot and the interaction between SIRT1 and FOXO1 by CO-IP.For Statistical Analysis.XIIResults:1.Comparison of basic measurement parameters: Compared with the model control group,the food intake(g)of the model vegetarian group has decreased(21.93±2.98 vs 30.91±2.18),the difference is statistically significant(t=8.76,P<0.01).Compared with the normal control group,the compound nutrition and vegetarian intervention in January showed a significant increase in body weight(g)(52.4±2.789 vs 23.74±1.24)and a significant increase in body mass index(kg /m2)(5.49±0.41 vs 2.83±0.2),liver wet weight(g)increased significantly(3.45±0.45 vs 1.04±0.17),liver index(%)increased significantly(6.56±0.55 vs 4.41±0.74),the differences were statistically significant(t values were 21.001,12.926,11.2 and 5.222,P<0.01).Compared with the normal control group in March,the compound nutrition vegetarian intervention significantly increased the body weight(g)of the model control group(59.87±4.63 vs 24.26±4.16),and the body mass index(kg/m2)increased significantly(6.55±0.48 vs 2.91±0.26),liver wet weight(g)increased significantly(4.42±0.39 vs 1.29±0.29),liver index(%)increased significantly(7.38 ± 0.39 vs 5.3 ± 0.47),the differences were statistically significant(t values were 12.796,14.899,14.41 and 7.647,P<0.01);compared with the model control group,the model vegetarian group's body weight(g)decreased significantly(50.28±2.37 vs 59.87±4.63),body mass index(kg/m2)decreased significantly(5.36±0.21 vs 6.55±0.48),liver wet weight(g)decreased significantly(3.14±0.64 vs 3.45±0.45),liver index(%)decreased significantly(5.3±0.4 vs 7.38±0.39),the differences were statistically significant Significance(t values were 4.12,5.048,8.634 and 8.356,respectively,P <0.01).2.Comparison of serum ALT,AST and CHOL levels: Compound nutritionalvegetarian intervention for 1 month,compared with the normal control group,the serum ALT(U/L)level of the model control group increased significantly(688.16±289.34 vs 90.14±68.6),serum CHOL(mmol/L)level increased significantly(5.39±1.21 vs 2.61±0.77),the differences were statistically significant(t value was 4.497 and 4.334,P<0.01);compared with the model control group,the model vegetarian group Serum ALT(U/L)level decreased significantly(150.96±54.77 vs 688.16±289.34),the difference was statistically significant(t=4.079,P<0.01).Compared with the normal control group in March,the combined nutrition and vegetarian intervention,the model control group's serum ALT(U/L)increased significantly(623.54±202.36 vs 103.55±25.51),and the serum CHOL(mmol/L)level increased significantly(7.3 ± 0.59 vs 3.46 ± 0.85),the difference is statistically significant(t values are 5.038 and 8.046,respectively,P<0.01);compared with the model control group,the serum level of ALT(U/L)in the model vegetarian group decreased significantly(55.58±11.61 vs 623.54±202.36),serum AST(U/L)level decreased significantly(154.78±33.6 vs 441.6±89.11),serum CHOL(mmol/L)level decreased significantly(4.29 ± 0.57 vs 7.3 ± 0.59),all differences Statistical significance(t values were 6.266,6.735 and 8.189,respectively,P<0.01).3.Pathology of liver tissue: pathology of liver tissue in normal control group and normal vegetarian group showed normal morphological structure of hepatocytes,no steatosis,inflammatory cell infiltration and fibrosis,etc.However,the disorder of liver cells in model control group showed obvious steatosis,Infiltration of interstitial inflammatory cells showed a small number of fibrous enlargements in the manifold area;hepatocyte steatosis and inflammation cell infiltration in the model vegetarian group were reduced.The liver tissue SAF scoreXIVshowed that the compound nutrition vegetarian intervention in January,compared with the normal control group,the model control group SAF score increased significantly(5.6 ± 1.14 vs0 ± 0),the difference was statistically significant(t=10.983,P<0.01)Compared with the model control group,the model vegetarian group had no significant difference in SAF score(5.4 ±1.14 vs 5.6± 1.14,t =0.277,P> 0.05);compound nutrition vegetarian intervention for 3 months,compared with the normal control group,model control The SAF score of the group increased significantly(6.8±0.84 vs 0±0),the difference was statistically significant(t=18.174,P<0.01);compared with the model control group,the SAF score of the model vegetarian group decreased significantly(4.2±1.1 vs 6.8±0.84),the difference is statistically significant(t=4.218,P <0.01).4.Comparison of SIRT1 mRNA expression level in liver tissue: Compound nutrition and vegetarian intervention for 1 month,compared with normal control group,SIRT1 mRNA expression level of model control group was significantly reduced(0.266 ± 0.051 vs 0.813 ± 0.097),the difference was statistically significant(t=11.167,P<0.01);compared with the model control group,the SIRT1 mRNA expression level of the model vegetarian group increased significantly(0.486 ± 0.1 vs 0.266 ± 0.051),the difference was statistically significant(t=4.372,P<0.01).Compared with the normal control group for 3months,the expression of SIRT1 mRNA in the model control group was significantly reduced(0.212 ± 0.077 vs 0.942 ± 0.294).The difference was statistically significant(t=5.368,P<0.01),compared with the model control group.Compared with the group,the SIRT1 mRNA expression level of the model vegetarian group increased significantly(0.503 ± 0.08 vs 0.212 ± 0.077),the difference was statistically significant(t = 6.278,P<0.01).5.Comparison of SIRT1 and AC-FOXO1 protein in liver tissue: Compound nutritional vegetarian intervention for 1 month,compared with normal control group,SIRT1 protein level in model control group was significantly reduced(0.094±0.067 vs 0.334±0.073),AC-FOXO1 protein level was obvious Increase(0.344±0.092 vs 0.118±0.081),the difference is statistically significant(t value were 5.432 and 4.126,P<0.01);compared with the model control group,the model vegetarian group SIRT1 protein level showed an upward trend(0.218±0.084 vs0.094±0.067),the AC-FOXO1 protein level showed a downward trend(0.118±0.081 vs 0.344±0.092),but the difference was not statistically significant(t values were 2.586 and 1.123,P>0.05).Compared with the normal control group,the compound nutrition vegetarian intervention in March,the model control group SIRT1 protein level was significantly reduced(0.124±0.097 vs 0.352±0.086),AC-FOXO1 protein level was significantly increased(0.328 ± 0.008vs0.12 ±0.042),The differences were statistically significant(t values were 3.928 and10.902,respectively,P<0.01);compared with the model control group,the model vegetarian group SIRT1 protein level showed an upward trend(0.194±0.059 vs0.124 ± 0.097),but the difference No statistical significance(t=1.38,P>0.05),AC-FOXO1 protein level decreased significantly(0.208±0.052vs0.328±0.008),the difference was statistically significant(t = 5.126,P <0.05).6.Interaction between SIRT1 and FOXO1: The results suggest that there is an interaction between FOXO1 and SIRT1.Conclusion:1.Compound nutrition and vegetarian diet can regulate liver lipid metabolism in Ob / Ob model mice,reduce enzyme levels,reduce inflammation,improve histopathology,and treat non-alcoholic fatty liver disease.2.The compound nutrition vegetarian diet upregulates the expression of SIRT1,thereby regulating the expression of its downstream target gene FOXO1 acetylation,activates the SIRT1 / FOXO1 signaling pathway to control the weight gain of Ob / Ob model mice,regulate lipid metabolism,and reduce liver fat accumulation and inflammation Improve histopathology. |
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