Effects of berberine and its combinations on adipogenesis

The objective of this study is to investigate the effects of berberine and its combinations (with evodiamine or amentoflavone) on adipogenesis and their transcriptional impacts during this process in vitro (cells) and in vivo (animals) with or without induction of second generation anti-psychotics (clozapine and risperidone).;When mouse 3T3-L1 preadipocytes were treated with berberine (1-64 μM) during differentiation, the results showed that berberine had a minor effect on cell viability during differentiation induced by differentiation medium. Quantitative Oil-Red-O staining measurements of 3T3-L1 cells which were grown in 1, 2, 4, 8 μM berberine-containing medium and were 8 days post differentiation induction, demonstration a reduction in fat droplets of 59.4%, 58.4%, 71.8% and 72.6% respectively. Treatment with 8 μM berberine for 8 days decreased the expression of PPARγ and C/EBPα, and increased expression of GATA-2 and GATA-3 both at the gene and protein levels.;When human white preadipocytes (HWP) were treated with berberine, evodiamine, and their combination during differentiation, the results showed that individually, berberine had little effect on HWP viability, but evodiamine greatly inhibited cell viability. However, in combination, the compounds showed only a slight inhibition of viability. Individually and in combination, treatment with 8 μM berberine or 4 μM evodiamine resulted in a major inhibition of HWP differentiation accompanied by an up-regulation of both GATA-2 and 3 mRNA and protein expression. However, the expression of both PPARγ and C/EBPα remained unchanged. Nonetheless, treatment of HWP with a combination of berberine and evodamine did not result in any additive or synergetic inhibition of the differentiation process.;When the effects of berberine on the adipogenesis of high-fat diet-induced obesity (FD) or normal diet (ND) mice and possible transcriptional impacts were investigated, the results demonstrated that in FD mice, berberine reduced mouse weight gain and food intake, and decreased the weight of epididymal fat and liver relative to body weight. Serum glucose, triglyceride, and total cholesterol levels were also reduced. These changes were accompanied with a down-regulation of PPARγ mRNA and protein expression and an up-regulation of both gene and protein expression of GATA-3. Berberine had no significant effect on weight gain, epididymal fat weight, food intake, serum glucose, triglyceride, total cholesterol levels in ND mice, and did not affect the kidney, spleen and liver weight in both ND and FD mice.;When mouse 3T3-L1 preadipocytes were treated with berberine during differentiation induced by differentiation medium with addition of clozapine or risperidone, the results showed that neither clozapine nor risperidone, alone or in combination with berberine had significant effects on cell viability. Eight days treatment with 15μM clozapine increased adipogenesis by 37.4% and 50 μM risperidone increased adipogenesis by 26.5% during 3T3-L1 cell differentiation accompanied by increased SREBP-1, PPARγ, C/EBPα, LDLR, and Adiponectin gene expression. More importantly, the addition of 8 μM berberine diminished the induction of adipogenesis almost completely accompanied by down-regulated mRNA and protein expression levels of SREBP-1-related proteins.;When mouse 3T3-L1 preadipocytes were treated with various concentration of berberine combined with amentoflavone during differentiation induced by differentiation medium and 15 μM clozapine, results showed that those combinations have no significant effects on cell viabilities, quantification of Oil-Red-O staining demonstrated that individually, amentoflavone did not inhibit adipogenesis significantly, however interestingly, the combination of 25μM amentoflavone with 1, 2, 4, 8 μM berberine produced higher significant inhibition of adipogenesis comparing to berberine alone, 8 μM berberine combined with 25μM amentoflavone inhibited adipogenesis by 97%. Realtime RT-PCR and Western Blot showed that amentoflavone enhanced the inhibitory effects of berberine on mRNA and protein expression of SREBP-1, PPARγ, and C/EBPα. (Abstract shortened by UMI.).