The Influence Of Pioglitazone Derivatives On3T3-L1Cells Function

objective：To further enhance curative effect of insulin sensitizers andreduce the drugs side effects, a new compound—CQMUHS-03wasdeveloped from pioglitazone. In order to provide a theoretical basis forCQMUHS-03in the treatment of type2diabetes, we investigated theinhibitory effect and the differentiation role of CQMUHS-03on3T3-L1cells in vitro, and discussed the possible mechanism at the same time.Method：（1）The3T3-L1cells were treated with CQMUHS-03(1×10^-8mol/L～1×10^-4mol/L) for24,48,72hours. MTT assay was used toinvestigate its inhibitory effect on the survival rate of3T3-L1cells.（2） Atfirst3T3-L1cell differentiation model was established. Then3T3-L1cellswere treated with CQMUHS-03at the begining of differentiation. On the2,4,6,8d during the differentiation, cells were stained with Oil Red O. Thentook cells pictures and detected the optical density at570nm.（3）3T3-L1cells were exposed to CQMUHS-03until them became mature fat cells.The changes of PPARγ gene expression were analyzed by Real-time PCR.（4）3T3-L1cells were treated with CQMUHS-03when been induced todifferentiation. On the2,4,6,8d during the differentiation, cells wereanalyzed by Western Blot to observe the expression of PPARγ protein.Results：（1） MTT assay showed that CQMUHS-03inhibit the growthof3T3-L1cells in a time and dose dependent fashion. Compared withnormal cells，CQMUHS-03significantly inhibited the growth of3T3-L1cells at1×10^-4～1×10^-5mol/L（P＜0.05）. On the other hand, pioglitazonepresent significant inhibitory effect at1×10^-4～1×10^-6mol/L（P＜0.05）. Compare with two compounds, pioglitazone inhibited the cells more potent（P＜0.05）.（2） The results showed pioglitazone promoted the cellsdifferentiation significantly （P＜0.05）. But the effect of CQMUHS-03todifferentiation was not conspicuous with control group.（3） Real-time PCRanalysis of PPAR γ showed that both CQMUHS-03（2.29times） andpioglitazone （5.12times） raised the PPARγ mRNA. Further more,pioglitazone was more obvious than CQMUHS-03（p<0.05）.（4） Westernblot analysis of PPARγ displayed that the protein of PPARγincreased infirst4days and then decreased, no matter pioglitazone or CQMUHS-03.The expression of PPARγ protein with pioglitazone also more significantthan CQMUHS-03（P＜0.05）.Conclusion：（1） CQMUHS-03has slight inhibitory effect to3T3-L1cells under1×10^-6mol/L and displays less toxicity than pioglitazone.（2）CQMUHS-03has slight improvement to the differentiation of3T3-L1cells.But pioglitazone improves the differentiation of cells significantly （P＜0.05）.（3） CQMUHS-03significantly increases PPARγ mRNA in3T3-L1differentiation but still less than pioglitazone.（4） CQMUHS-03significantly raises the expression of PPARγ protein in differentiation of3T3-L1cells but less than pioglitazone.