Analysis Of Related Factors Of Pulmonary Function Changes In Children With Non-alcoholic Fatty Liver Disease And Tilianin Protects Against Nonalcoholic Fatty Liver Disease In Early Obesity Mice

Objective: To study the changes in pulmonary function and its influencing factors in children with non-alcoholic fatty liver disease.To explore the functions and possible mechanisms of Tilianin on NAFLD in early obesity mice.Methods: Lung function indexes(VCMAX,FVC,FEV1,PEF,FEV1/FVC,FEF25,FEF50,FEF75,MMEF25-75)were measured in children with non-alcoholic fatty liver disease and normal control group.Physical development indicators(BMI,waist circumference),liver function(ALT,AST,GGT),lipid metabolism and glucose metabolism indicators(TC,TG,HDL-C,LDL-C,blood glucose,serum insulin,HOMA-IR),cytokines(IL-1β,IL-18)were detected.A high-fat high-carbohydrate(HFHC)diet was used to feed obese mice,in order to establish juvenile obese mouse model of nonalcoholic fatty liver disease.Tilianin was administered daily at a dose of 10 or 20mg/kg.The serum of mice was collected for detection of(ALT,AST,GGT).Liver tissue was homogenized to determine the levels of(TC,HDL-C,LDL-C),The above biochemical parameters were measured using the kits,The levels of TNF-α,IL-1β,IL-6 and IL-18 in serum and liver were measured using the commercial enzyme-linked immunosorbent assay(ELISA)kits.The protein expression levels of ACC1,FAS,SREBP-1c,LXRα,Pro caspase-1,active caspase-1 and NLRP3 protein levels in the liver were analyzed by western blotting.The expression of F4/80 was analyzed by immunofluorescence.ROS concentration in mice detected using DHE probe.4-HNE expression was analyzed by IHC analysis.The above indicators were compared and analyzed.Results: A total of 35 children with NAFLD(15 females and 20 males)were enrolled,among whom 28 had nonalcoholic fatty liver disease(NAFL)and 7 had nonalcoholic steatohepatitis(NASH).Thirty healthy children were recruited as the control group,including 13 girls and 17 boys.VCMAX and FVC in the control group were lower than those in the NAFLD group,and the differences were statistically significant.PEF,FEV1/FVC,FEF75,and MMEF25-75 in the control group were higher than those in the NAFLD group,and the differences were statistically significant.There was no significant difference in FEV1,FEF 25,FEF50 between the control group and the NAFLD group.There were no significant differences in VCMAX,FVC,PEF,FEV1,FEF25,FEF50 between NAFL group and NASH group.FEV1/FVC and MMEF25-75 in NAFL group were lower than those in NASH group,and the differences were statistically significant.There was a moderate negative correlation between FEF50,FEF75 and AST(r=-0.339,r=-0.317),There was a strong negative correlation between MMEF25-75 and AST(r=-0.528,P<0.001),triglyceride had a moderate negative correlation with FEV1(r=-0.361,P=0.003),and a moderate negative correlation with PEF(r=-0.347,P=0.005).FEV1/FVC had a moderate negative correlation with HOMA-IR(r=-0.395,P<0.001).Moderate negative correlation was found between PEF and HOMA-IR(r=-0.374,P=0.002).The decrease in the small airway dysfunction markers FEF50,FEF75 and MMEF25-75 showed a moderately negatively correlated with HOMA-IR,and the correlation coefficients were(-0.418,-0.361,-0.417),There was a moderate negative correlation between the decrease of MMEF 25-75 and the levels of IL-1β and IL-18,and the correlation coefficients were(-0.412,-0.417).HFHC-fed mice gained weight,increased liver index.The liver showed hepatocyte ballooning,inflammatory infiltration,and steatosis.Elevated levels of ALT,AST,GGT,LDL-C and TC and reduced the HDLC level were found in HFHC-fed mice.Administration of tilianin significantly reduced these impairments.We further evaluated proteins related to lipid metabolism and observed that LXRα,SREBP-1c,FAS and ACC1 expression were blunted following tilianin administration.In addition,tilianin suppressed ROS overproduction and lipid peroxide 4-Hydroxynonenal expression,ascribed to its oxidative stress-modulating capacity.Tilianin also reversed the increase in F4/80 expression and proinflammatory cytokine levels.Of note,tilianin administration resulted in decreased protein levels of active caspase-1 and NOD-like receptor protein 3(NLRP3)in HFHC-fed mice.Conclusions:(1)Compared with normal children,In NAFLD children,lung volume is increased,while FEV1/FVC,PEF,and small airway function are decreased.A complex correlation was seen between these changes and HOMA-IR,BMI,TG,AST,LDL-C,IL-1β,IL-18.(2)Insulin resistance and elevated serum IL-1β and IL-18 levels may be the main correlates of abnormal lung function in children with NAFLD.(3)Tilianin was able to regulate SREBP-1c expression via LXRα and attenuated hepatic lipid accumulation.(4)Tilianin ameliorates NAFLD disease process in early obese mice by inhibiting ROS production and attenuating inflammatory responses as well as oxidative stress,which may involve,in part,inhibition of NLRP3 inflammatory vesicle activation.