吡格列酮衍生物CQMUHS-03对3T3-L1细胞增殖分化的影响  

Influence of pioglitazone derivatives CQMUHS-03 on 3T3-L1 cells proliferation and differentiation

作　　者：刘菲[1] 舒丽[1] 胡湘南[1] 孙文娟[1] 

机构地区：[1]重庆医科大学药理学教研室,重庆市生物化学与分子药理学重点实验室,重庆400016

出　　处：《中国药理学通报》2014年第1期66-71,共6页Chinese Pharmacological Bulletin

基　　金：重庆市自然科学基金资助项目(No CSTC 2006BB5286)

摘　　要：目的研究吡格列酮衍生物CQMUHS-03对3T3-L1细胞增殖及诱导分化的影响,为CQMUHS-03的临床应用提供理论基础。方法 (1)不同浓度药物处理细胞,于24、48、72h,MTT法检测CQMUHS-03对3T3-L1前脂肪细胞增殖的影响。(2)建立3T3-L1细胞诱导分化模型,药物处理后经油红O染色,拍照后并测定570 nm处光密度值以确定有效药物干预浓度。(3)Western blot测定药物对PPARγ蛋白表达的影响。(4)Real-time PCR分析PPARγ基因表达。结果(1)经24、48、72 h MTT检测,1×10-6mol·L-1浓度以下的CQMUHS-03对3T3-L1细胞增殖的抑制作用弱,IC50为3.33×10-4mol·L-1,高于吡格列酮(2.91×10-4mol·L-1)。(2)油红O染色测定结果显示吡格列酮促进3T3-L1前脂肪细胞分化,而CQMUHS-03对3T3-L1前脂肪细胞的分化促进作用不明显。(3)Real-time PCR检测显示诱导分化8 d,吡格列酮组PPARγmRNA表达上调5.12倍,CQMUHS-03组PPARγmRNA表达上调2.29倍。(4)Western blot结果显示,CQMUHS-03使PPARγ的表达增高。结论吡格列酮衍生物CQMUHS-03对3T3-L1细胞抑制增殖作用弱于吡格列酮,对细胞分化无明显促进作用,能上调PPARγ基因和蛋白表达。Aim To investigate the inhibitory effect and the differentiation role of CQMUHS-03 on 3T3-L1 cells, and the underlying mechanism for the clinical use . Methods （1） 3T3-L1 cells were treated with CQMUHS-03 （1 × 10-8mol · L^-1 - 1 × 10^-4 mol · L^-1 ） for 24, 48, 72 hours. MTT assay was used to in- vestigate its inhibitory effect on 3T3-L1 cells. （2） 3T3-L1 cells were treated with CQMUHS-03 at the be- ginning of differentiation, then cells were stained with Oil Red O, cell pictures were taken, and the optical density at 570nm was detected. （3） When cells turned into mature fat cells, PPAR＇y gene expression was ana- lyzed by Real-timePCR. （4）On2 nd, 4 th, 6 th, 8 th during the differentiation, the expression of PPARγ was analyzed by Western blot. Results （ 1 ） MTT as- say showed that CQMUHS-03 had a slightly inhibitory effect on cells under 1 × 10^-6 mol · L^-1 （ IC50 = 3.33 × 10^-4 mol · L^-1）. （2）Pioglitazone promoted the cell differentiation significantly （P 〈 0.05 ）. However, the effect of CQMUHS-03 on differentiation was not con- spicuous in control group. （3） Real-time PCR analysis of PPARγ showed that both CQMUHS-03 （2.29 times） and piogtitazone （5.12 times） raised the PPARγ mR- NA. （4） Western blot analysis of PPARγ displayed that the protein of PPARγ increased in the first 4 days and then decreased, no matter pioglitazone or CQMU- HS-03. The expression of PPARγ protein with pioglita- zone was also more significant than that with CQMUHS- 03 （P 〈 0. 05 ）. Conclusions CQMUHS-03 has more slightly inhibitory effect on 3T3-L1 cells than pioglita- zone, and it does not improve the differentiation of cells significantly. CQMUHS-03 significantly increases PPARγ mRNA and the expression of PPARγ protein in differentiation of 3T3-L1 cells, but the effect is less than that of pioglitazone.

关 键 词：吡格列酮 CQMUHS-03 3T3-L1前脂肪细胞 增殖 分化 PPAR3 

分 类 号：R-332[医药卫生] R329.24