The Experimental Research And Clinical Observation Of The Effects Of PPAR-γ Agonist On Human Uterine Leiomyoma

Objective: The aim of the present study is how to get the uterine leiomyoma cells in vitro which have the similar bionomics to the leiomyoma cells in human body, and is to discuss the senses of ER, PR, PPAR-γand TGF-β3 expressions in the uterine leiomyoma tissues, normal autologous unaffected myometrium and the cultured uterine leiomyoma cells in vitro.Methods: Human uterine leiomyoma tissues were dissected and cultured. Characterization of the cultured cells was examined using immunostaining with monoclonal antibodies to a muscle-specific protein desmin. RT-PCR was used to detect the mRNA expressions of ER , PR , PPAR-γ and TGF-β3 and Immunofluorescence staining was used to detect the protein expressions of ER, PPAR-γ and TGF-β3 in the cultured uterine leiomyoma cells, uterine leiomyoma tissues and the normal autologous unaffected myometrium.Results: The uterine leiomyoma cells were got by using collagenase dissection and were confirmed by immunostaining with monoclonal antibodies to a muscle-specific protein desmin. RT-PCR analysis revealed that ER,PR,PPAR-γand TGF-β3 mRNA expressions have no significant changes, between the cultured cells in vitro and the uterine leiomyoma tissues (P > 0.05) ,however, ER, PR, PPAR-γ and TGF-β3 mRNA expressions in the normal autologous unaffected myometrium were decreased to compare with the cultured cells and the uterine leiomyoma tissues (P < 0.05 ) . Immunofluorescence staining demonstrated that ER, PR, PPAR-γ and TGF-β3 protein expressions were similar to those mRNA expressions.Conclusions: The uterine leiomyoma cells can be got through the method of collagenase dissection which have the similar bionomics to the uterine leiomyoma cells in human body. And the genes of ER, PR, PPAR-γ, TGF-β3 have important roles in the pathomechanism of human uterine leiomyoma. Objective: The aim of our study was to investigate the effects of rosiglitazone on uterine leiomyoma cells. Gene expressions of PPAR-γ,TGF-β3,Bcl-2,Bax and cell proliferation as well as cell apoptosis were investigated in uterine leiomyoma cells in vitro.Methods: Human uterine leiomyoma tissues were dissected and cultured. Cells were divided into 5 groups: one control group and other four groups with different concentrations of rosiglitazone (10^-7, 10^-8,10^-9 and 10^-10 mol/l). Cells were cultured for 72 hours in serum-free Dulbecco's modified Eagle's medium (DMEM).MTT reduction assay was used to detect the cell proliferation. RT-PCR was used to detect the mRNA expression of PPAR-γ and TGF-β3. Immunofluorescence staining was used to detect the protein expressions of PPAR-γand TGF-β3. TUNEL apoptosis staining was used to detect the percentage of cells apoptosis.Results: MTT reduction assay indicated that the treatment with rosiglitazone (from 10^-7 to 10^-9mol/l) resulted in an inhibition of the cell growths after 72 hours (P < 0.05),and there were no significant difference between each other in the treatment of 36h(P > 0.05). RT-PCR analysis revealed that excepted the trail group for 10^-7 mol/1 ,the expressions of PPAR-γand Bcl-2 mRNA in the other trail groups were no significant difference compared with the control groups at the treatment of 36h. The expressions of TGF-β3 , Bax mRNA in all the trail groups have no significant difference compared with the control group at the treatment of 36h. 10^-7 mol/l rosiglitazone significantly affected the gene expression at 72-hour: PPAR-yand Bax mRNA expression was up-regulated and TGF-β3 and Bcl-2 mRNA was down-regulated and rosiglitazone at the concentration of 10^7- mol/l affected theses most effectively(P< 0.01). Immunofluorescence staining demonstrated that treatment with 10^-7 mol/l rosiglitazone resulted in the significant changes of PPAR-γ and TGF-β3 protein expressions compared with the other treatment groups and the control group at 72 h (P < 0.01). PPAR-γprotein expression was up-regulated and TGF-β3 protein expression was down-regulated by rosigltaizone. The percentage of apoptosis increased with treatment dose at 72 h. Treatment with 10^-7 mol/l rosiglitazone resulted in the highest percentage of apoptosis compared with the other treatment groups and the control (P < 0.01). All the effects of rosiglitazone on uterine leiomyoma cells were dose-dependent and time-dependent in vitro.Conclusions: The present study demonstrates that the PPAR-γ activator, rosigltaizone, inhibits the cell proliferation and increase the cell apoptosis of uterine leiomyoma cells in vitro. The effect maybe through regulations of PPAR-γ, TGF-β3 , Bcl-2 and bax expressions in vitro. Objective: The aim of our study was to investigate the effects of AVANDIA ( rosiglitazone ) on human uterine leiomyoma. Gene expressions of PPAR-γ ,TGF-β3, Bcl-2 and Bax as well as cell apoptosis were investigated in uterine leiomyoma.Methods: The patients for uterine leiomyoma were divided into the control group and the trail groups . Hepatic function, renal function, fasting blood glucose, estrogen and progestogen in serum were detected for the control group patients and the trail group patients before and after treatment. B ultrasound wad used to detect the uterine leiomyoma's volume before and after treatment in trail group. RT-PCR was used to detect the mRNA expression of PPAR-γ , TGF-β3, Bcl-2 and Bax of the control group or the trail group's uterine leiomyoma cells respectively . Immunofluorescence staining was used to detect the protein expressions of PPAR-γ and TGF-β3 of the control group or the trail group's uterine leiomyoma cells respectively . TUNEL apoptosis staining was used to detect the percentage of cells apoptosis.Results: Hepatic function, renal function, fasting blood glucose, estrogen and progestogen in serum were no significant difference between the trail group and the control group(P > 0.05).And there had no statistical difference between the before the treatment of AVANDIA and the after the treatment in the trail group(P > 0.05). B ultrasound assay indicated that the treatment with AVANDIA resulted in an inhibition of the uterine leiomyoma volume expansion, however the difference was no statistical significance(P> 0.05) . RT-PCR analysis revealed that AVANDIA significantly affected the gene expression : PPAR-γ mRNA and Bax mRNA expressions were up-regulated and TGF-63 and Bcl-2 mRNA expressions were down-regulated (P < 0.05). Immunofluorescence staining demonstrated that treatment with AVANDIA resulted in the significant changes of PPAR-γ and TGF-β3 protein expressions compared with the control group (P < 0.05). The percentage of apoptosis increased with treatment of AVANDIA compared with the control group (P < 0.01).Conclusions: The present study demonstrates that the PPAR-γ activator, AVANDIA, inhibits the cell proliferation and increased the cell apoptosis in human uterine leiomyoma.