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

Aim To investigate the inhibitory effect and the differentiation role of CQMUHS-03 on 3T3-L1cells,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- 4mol ·L- 1) for 24,48,72 hours. MTT assay was used to investigate its inhibitory effect on 3T3-L1 cells.( 2)3T3-L1 cells were treated with CQMUHS-03 at the beginning 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,PPARγ gene expression was analyzed by Real-time PCR.( 4) On 2 nd,4 th,6 th,8th during the differentiation,the expression of PPARγwas analyzed by Western blot. Results( 1) MTT assay showed that CQMUHS-03 had a slightly inhibitory effect on cells under 1 × 10- 6mol·L- 1( IC50= 3. 33× 10- 4mol·L- 1).( 2) Pioglitazone promoted the cell differentiation significantly( P 0. 05). However,the effect of CQMUHS-03 on differentiation was not conspicuous in control group.( 3) Real-time PCR analysis of PPARγ showed that both CQMUHS-03( 2. 29 times)and pioglitazone( 5. 12 times) raised the PPARγ mRNA.( 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 CQMUHS-03. The expression of PPARγ protein with pioglitazone 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 pioglitazone,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.