Olanzapine Induced SREBPs-related Adipogenesis And Co-administration In3T3-L1Cells

Background:Antipsychotic drugs are the first choice for treatment psychosis, mainly divided into two categories, the first generation of atypical drugs (FGAs) and second-generation atypical drugs (SGAs). The FGAs not only have no significant effect on negative symptoms of schizophrenia, which causing drug-induced extrapyramidal side effects, but also impede the restoration of social functions. The SGAs both have antagonism on DA receptors and5-hydroxytryptamine (5-HT) receptor without causing drug-induced extrapyramidal side effects. In addition, there are certain side effects such as clozapine-induced agranulocytosis which incidence as high as1‰～3‰which is50to100times that of other antipsychotic drugs, resulting in a significant clinical application is limited. Olanzapine as one representative of the SGAs, has more affinity on5-HT receptor than the DA receptor, and selectively acts on the limbic system in the brain and mental activity. It plays a good role in treatment the negative symptoms of schizophrenia and refractory schizophrenia without induced agranulocytosis. However, Olanzapine causes some adverse reactions, such as lipid metabolism disorders, dyslipidemia associated with obesity, diabetes, cardiovascular diseases and disorders related to inflammation, especially dyslipidemia is common. Now there is no drugs could replace Olanzapine in the clinical, therefore finding the drug which combined with Olanzapine can improve antipsychotic-induced abnormal lipid metabolism drugs becomes very urgent. The lipid metabolism disorders specific mechanisms of Olanzapine is not clear, it has been reported that side effects related with the H1receptor, lipogenesis factors and SREBPs pathways. Recent it is reported that Olanzapine lipid metabolism disorders may also have feedback regulation. Statins, lowering the first-line drugs, through competitive inhibition of HMG-CoA reductase and reduce cholesterol synthesis in the liver, but also increase LDLC receptor number and activity, increased serum cholesterol clearance, further reducing TC levels. Speculated that statins whether could inhibit Olanzapine-induced HMG-CoA reductase enhanced through SREBPs relevant pathway to interfere with lipid metabolism Olanzapine caused concern.In addition, Berberine which is from Berberis, Coptis rhizome extract plant a quaternary isoquinoline alkaloids, mainly used for intestinal inflammation and fever. It also has antibacterial, diarrhea, anti-arrhythmic, anti-hypertensive, drop lipids and other effects.Many literatures reported that Berberine also had more significant cholesterol-lowering effect, the exact mechanisms are not clear. It has been reported Berberine inhibits the adipocyte differentiation in lipogenesis of3T3-L1mouse embryonic fibroblasts, human white preadipocytes and high fat diet-induced obese mouse model through SREBPs pathway (including PPARy and C/EBPa). Therefore, Berberine is combination with Olanzapine whether could interfere with3T3-L1mouse embryonic fibroblasts lipid metabolism mediated by Olanzapine and thus applied to the clinical, is worth studying.Objective:Olanzapine-induced lipid metabolism mature adipocytes model was established and investigated the effect of statins and Berberine intervention and molecular mechanisms.Methods:This study investigated the impact of DMSO, Olanzapine, Simvastatin, Atorvastatin, Berberine on3T3-L1cell proliferation through MTT assay, in order to identify these drugs have a lower concentration range of cytotoxicity. To examine cell morphology, oil red O staining lipid droplets, detection kit TG, TC levels, RT-qPCR analysis and Western blottingting four aspects of Olanzapine-induced mature adipocytes model, and select the appropriate Olanzapine concentration. From the above four aspects to study statins and Berberine whether interfere the lipid metabolism disorders mediated Olanzapine and related molecular mechanisms.Results:1. To establish the lipid metabolism mediated by OlanzapineOlanzapine can promote the formation of lipid droplets in fat cells, and the effect is dose-dependent in the range of0-10μmol/L concentration, outside this range, the effect is not obvious. TG, TC amount, the level of gene transcription and protein expression is the same with the phenomenon of lipid droplets.2. The intervention of Simvastatin with lipid metabolism disorders caused by Olanzapine2.5μmol/L and5μmol/L Simvastatin could change the cell morphology and the number of lipid droplets, TG, TC quantity. The analysis of gene transcription and protein expression levels were further validated Simvastatin may interfere with lipid metabolism disorders caused by Olanzapine.3. The effect of Atorvastatin on Olanzapine-induced lipid metabolism disorders5μmol/L Atorvastatin basically could change the cell morphology, the number of oil red O staining lipid droplets and the number of TG, TC, gene transcription and protein expression analysis.4. The effect of Berberine on lipid metabolism disorders mediated by OlanzapineFrom1.25μmol/L Berberine could clearly alter the cell morphology and the oil red O staining lipid droplets, TG, TC volum. Further to analyze gene transcription and protein expression levels Berberine may interfere with lipid metabolism disorders caused by Olanzapine.Conclusion:This study confirms the Olanzapine mediated3T3-L1adipocyte lipid metabolism and establish cell model. To find Simvastatin and Berberine combined with Olanzapine through this model, can interfere with lipid metabolism side effects of olanzapine which is the representative of SGAs, provide the basis for clinical drug combination.