High Fat Diet And Berberine On Nonalcoholic Fatty Liver Disease Were Growth And The Effects Of Insulin-like Growth Factor Expression

Objective1.To investigate the expression of IGF-1and IGFBP-3in nonalcoholic fatty steatosis hepatocyte models induced by oleic acid.2.To investigate the effects of berberine on the expression levels of IGF-1and IGFBP-3in the nonalcoholic fatty liver disease cell model induced by oleic acid.3.To investigate the effects on growth and expression of IGF-1level in young rats induced by high-fat feeding, and illustrate relationship between growth failure in young rats with non-alcoholic fatty liver disease and IGF-1turbulence of serum and liver tissue.MethodsUpon oleic acid inducing immortalized human hepatocyte(IHH), NAFLD cell model was established. Oil red O staining and intracellular triglycerides were detected for observing the situation of intracellular hepatocyte steatosis. The IHHs were divided into control group、oleic acid-treated group, Which the control group cultured in DMEM/F12medium, NAFLD group were treated with0.5mmol/L oleic acid treated for72h. The expression levels of IGF-1、IGFBP-3mRNA and protein were measured by RT-PCR、Western-blotting and immunofluorescent staining.IHH cells were divided into control group, NAFLD group, berberine-treated group. The expression of IGF-1and IGFBP-3mRNA and protein were measured by RT-PCR. Western-blotting and immunofluorescent staining.Sixty SD young rats of3weeks old were randomly divided into three groups, normal group(N)、NAFLD group、berberine group(Ber). All the rats were fed under Specefic pathogen Free(SPF) condition for5weeks, the body weight, the nose-tail length were checked. To confirm the NAFLD young rats models were duplicated successfully, the rats in both groups were performed HE staining. serum biochemical indicators and intracellular TG detecting, the levels of serum IGF-1and IGFBP-3. The expression of mRNA and protein of IGF-1and IGFBP-3in liver tissue were measured by RT-PCR and Western-blotting methods.ResultsThe IHH steatosis model was established successfully with the treatment of0.5mmol/L oleic acid.Oil red O staining showed that the contents of intracellular lipid droplets increased significantly, and the levels of intracellular TG increased to 275.7(ug/mg protein) from150.2(ug/mg protein)(P<0.01). Compared with the control group, the expression levels of IGF-1and IGFBP-3mRNA and proteins significantly decreased in the oleic acid-treated group(P<0.01)。After treatment with lOumol/L berberine, oil red O staining showed that the contents of intracellular lipid droplets decreased, and the levels of intracellular TG decreased to236.1(ug/mg protein) from275.7(ug/mg protein)(P>0.05). the expression levels of IGF-1and IGFBP-3mRNA and proteins reverted to the levels of the control group after treatment with lOumol/L berberine.The typical pathological change of NAFLD was found in NAFLD groups with HE stained results induced by high-fat feeding for5weeks. Compared with N group, TC、 CRP、ALT levels and liver TG in NAFLD group were significantly elevated(P<0.01). After berberine intervention, the levels of serum TC、ALT decreased significantly. The body weigh、the nose-tail length in NAFLD group were significantly decreased (P<0.01),while their levels in berberine group decreased the NAFLD group (P>0.01). The lengthof the femoral and the tibial were lower than that in N group(P>0.05), these reduction were not significant, and their levels in berberine group have no statistical significantly compared with the NAFLD group(P>0.05). The femoral BMD and BMC in NAFLD group were lower than that in N group(P<0.01), while their levels in berberine group were significantly increased compared with the NAFLD group(P<0.01). The serum IGF-1and IGFBP-3levels in NAFLD group were significantly lower than those in N group, as well as the liver tissue IGF-1and IGFBP-3mRNA and protein in NAFLD group, while their levels reverted to the levels of the control group after treatment with berberine(P<0.01). The serum GH level in NAFLD group was lower than that in N group, but this reduction was not significant(P<0.01).ConclusionThe expression of hepatocyte IGF-1and IGFBP-3in nonalcoholic fatty steatosis models were decreased, which may provide experimental basis for the further mechanism study of some non-alcoholic fatty liver disease children with growth failure.Berberine can improve the expression levels of IGF-1and IGFBP-3in the nonalcoholic fatty liver disease cell model.There was a positive correlation of serum IGF-1with the nose-tail length and weight. The growth restriction of young rats induced by high-fat feeding may be related to the decressed expression of IGF-1. After berberine intervention, the weight, body length have no significant changees, but the expression of serum IGF-1、IGFBP-3levels and BMD、BMC were improved the role of the pups bone density, bone mineral content.