Farnesoid Receptor X On Lipid Metabolism Of SD Obese Offspring And Serum Biomarkers In Obese SD Offspring

PART I Effects of berberine on high-fat diet induced obese SD offspring through farnesoid receptor X singalling pathway Background and purpose: Clinical and animal studies have shown that berberine can improve metabolic disorders in obesity and its associated metabolic syndrome,non-alcoholic fatty liver disease and other diseases.Berberine reduces lipid synthesis and increases fatty acid oxidation,so that improves lipid metabolic disorders.The effects of berberine on lipid metabolism and other internal environment stability in young animal models have rarely been reported.The nuclear receptor farnesoid receptor X signaling pathway is involved in the pathophysiological processes of obesity,metabolic syndrome and other diseases.In addition to regulating bile acid metabolism,farnesoid receptors are involved in many aspects of hepatic lipid deposition.The role of the liver farnesoid receptor in metabolic effects induced by berberine has been poorly undertanded.High-fat diet induced obese SD offspring can be used as a laboratory research model for childrenand adolescents obesity.In this study,three models,including high fat indued obesity rats,palmitic acid/oleic acid-induced primary hepatocytes or Hep G2 cell steatosis,were applied to study the effect of BBR on lipid metabolism and the role of farnesoid receptor signaling pathway..Methods: Obese SD offspring were established through a persistent high-fat diet before conception,during pregnancy,lactation time,and after weaning.After weaning,rats were divided into two subgroup,berberine group and sodium carboxymethylcellulose control.group.Data about body weight,glucose and lipidmetabolism,hepatic lipid metabolism at 43 d were collected for analysis.HepG2 cells and rat primary hepatocytes were induced by oleic acid/palmitic acid for lipid accumulation.The farnesoid receptor si RNA were usted to interfered with primary hepatocytes and its inhibitor Z-Guggulsterone were applied to interfered with Hep G2 cells,amied to observe the effect of berberine on liver cell lipd metabolism.Results: The body weight,serum triglyceride,cholesterol,HOMA-IR,liver lipid deposition,etc.of rats fed with high-fat diet were significantly higher than those of the control group.The body fat content in the high-fat group exceeded that of the normal group.After 200 mg/kg body weight of berberine was continuously administered for 3weeks,the body weight was decreased compared with the non-intervention obese group.Body fat content,blood cholesterol,and liver triglyceride were all decreased.Hep G2 cells were adminstrated with 1.0 mmol/L oleic acid and 0.5 mmol/L.L-palmitic acid co-cultured for 24 h.lipid droplets accumulation were detected by oil red staining.Triglyceride content per cell were increased.Fatty acid oxidation-related molecules expression were decreased.Primary rat hepatocytes were successfully inoculated,the same ratio,concentration of oleic acid / palmitic acid.After 24 hours,the cells were induced by lipids.The deposition of lipid droplets were reduced after berberine adiminstrated.Triglyceride content in cell were reduced too.The expression of fatty acid oxidation-related molecules were increased inthe berberine intervention group.These characterization were also shown in the primary hepatocytes.Primary hepatocyte were treated with FXR si RNA,berberine,oleic acid/palm simultaneously.The mRNA and protein expression levels of FXR were significantly decreased,and the expression of FXR targetgene SHP was aslo down-regulated,and the expression of CYP7a1,suprressed by FXR,was up-regulated.The effect of berberine on lipid metabolism was weakened by FXR si RNA adminstration.Hep G2 cell line was treated with Z-Guggulsterone and berberine.After oleic acid/palmitic acid was cultured simultaneously,the effect of berberine on lipid metabolism was reduced,similar to primary hepatocytes treated with FXR si RNA.Conclusion: Obesity in SD offspringcan be induced by persistant high-fat diet.Body fat percentage,hepatic lipid deposition,blood glucose were increased in the obese offspring.Berberine can improve lipid/glucose metabolism in the obese offspring,such as blood glucose and lipid indexes,hepatic lipid deposition,etc..FXR si RNA and the FXR inhibitor retards the lipid-regulating effect of berberineon primary hepatocytes and HepG2 cell line.Berberine hydrochloride may be used for the treatment of children and adolscents obesity.Farnesoid receptor X in livermay be one of potential mechanisms for berberine effect on lipid metabolism.PRAT ? Characteristics of Wnt1-induced Signaling Protein1 and Fibroblast Growth Factor 1 in Obese SD Rats and youth Background and purpose: Wnt1 inducible signaling pathway protein-1(WISP1),a member of the CCN family,is considered as a potential target for obesity and type 2diabetes.WISP1 regulates low-grade inflammation in obese mice,and WISP1 levels are associated with obesity and adult type 2 diabetes.And another obesity-related bioactive marker,fibroblast growth factor 1(FGF1),regulates glucose and lipid metabolism in obese mice.Serum FGF1 levels increased in adults with type 2 diabetes.In this study,serum levels of WISP1 and FGF1 were measured in obese young SD rats and youth,and the relationship between WISP1 and interleukin 18(IL-18)was explored.Changes of serum FGF1 levels in berberine interfered SD rats and lifestyle intervented obese youth and their relationship with glucose and lipid metabolism.Methods: Forty-four obese children and adolescents(5 to 15 years old)were selected.At the same time,44 healthy children with normal weight at the child health clinic were used as controls;39 obese children and adolescent after 6 months of controlled diet and daily physical activity with weight loss above 5% were included in FGF1-related follow-up studies.All subjects were recorded for their physique,glucose and lipid metabolism and other laboratory parameters.Serum levels of FGF1,WISP1 and IL-18 were determined by enzyme-linked immunosorbent assay.SD rats obtained from the first part of the study,includingthe high-fat diet-induced obese group,the berberine intervention group,and the normal diet group were collected for fastingserum samples,to detectof FGF1,WISP1 and IL-18 levels.Results: WISP1 levels in obese young children were higher than those in controls(1364.08±18.69 vs 1735.44±15.29 pg/m L).WISP1 levels in obese SD rats and obese youth were higher than those in controls;WISP1 levels were positively correlated with body mass index(BMI)and BMI-z(r = 0.392,P = 0.008;r = 0.474,P = 0.001,respectively);the circulating IL-18 levels were increased in obese children(295.87 ±13.30 vs 1229.06 ± 29.42 pg/m L),and were also increased in obese SD rats;in the obese youth,serum WISP1 levels were correlated with IL-18(r = 0.542,P = 0.000),and adiponectin(r = 0.585,P = 0.000).There was a positive correlation(r = 0.592,P = 0.000)between serum WISP1 and IL-18 in obese SD rats.Multivariable stepwise linear regression analysis showed that after adjusting for BMI,waist circumference,fasting insulin,homeostatic model assessment of insulin resistance(HOMA-IR)and hemoglobin A1c(HbA1c)in obese children and adolescents,high-level serum IL-18 was the main determinant of high-level serum WISP1(? = 0.542,P = 0.000).Serum FGF1 levels were increased in obese SD rats and youth.Serum FGF1 levels in obese children were positively correlated with BMI and waist circumference(WC)(r = 0.377,P = 0.012;r =0.301,P = 0.047,respectively).Regression analysis showed that serum FGF1 levels in obese young infants were significantly associated with HbA1 c and HOMA-IR(? = 0.371,P = 0.008;? = 0.323,P = 0.021,respectively);The weight loss after intervention(2.34±0.11kg)was accompanied by a significant decrease in the serum FGF1 level(7.23±0.44pg/ml).Serum FGF1 levels of obese SD rat pups experienced a decrease in body weight after berberine intervention were decreased compared with the control group;the decrease of FGF1 were significantly related to HOMA-IR and low-density lipoprotein cholesterol(LDL-C)(? = 0.277,P = 0.020;? = 0.474,P<0.001;? = 0.320,P= 0.008,respectively).Conclusion:Serum levels of WISP1 and FGF1 are elevated in obese rats induced by a high-fat diet and obese youth.Increased WISP1 and FGF1 may increase the risk ofinsulin resistance;WISP1 may affect the metabolism of glucose and lipids in obese children and adolescents by IL-18 regulation;obese SD rats after berberine intervention and obese youth after lifestyle intervention have decreased serum FGF1 levels.This reduced serum FGF1 level is associated with improved insulin resistance after weight loss in obese individuals.WISP1 and FGF1 may have potential therapeutic effects on insulin resistance in obese children and adolescents.