Intense Pulsed Light in Effects of TGF-β1 Induced on the Proliferation of Fibroblasts and Intervention of ERK Inhibitor
|School||Central South University|
|Course||Dermatology and Venereology|
|Keywords||Intense pulsed light Transforming growth factor β1 Fibroblast ERK signaling pathway Cell proliferation|
Purpose. Observation of the transforming growth factor-p1 into cultured human skin fibroblast proliferation. Explore intense pulsed light in the transforming growth factor-p1-mediated fibroblast proliferation and extracellular signal-regulated kinase (ERK) signaling pathway. Method. Circumcision resection the normal adolescent foreskin organization, isolated and cultured in vitro normal human skin primary fibroblasts, keratin and Ⅰ collagen type III staining of fibroblasts identification. The experiment was divided into three parts: (1) Experiment: TGF-β1 concentration treatments, TGF-β1 treated separately with different concentrations of the constituent fiber cells, using methyl thiazolyl tetrazolium salt (MTT) assay to detect the the cell proliferation situation; (2) Experiment II: divided into ① TGF-β1 concentration group: 10.0ng/ml 5.0ng/ml and 1.0ng/ml and 0ng/ml ② The time group: giving 10.0ng/ml TGF-β1 stimulate cell protein extracted at different times Omin, 15min, 30min, 60min and 120min time points, respectively. Application protein immunoblot assay (western blot) groups phosphorylated extracellular signal-regulated kinase (p-ERK) changes. (3) experiment: the control group TGF-β11.0ng/ml 10.0ng/ml, the IPL irradiation dose 72J/cm2 group, 72J/cm2 TGF-β110.Ong/ml of PD98059 72J/cm2 group, PD98059 TGF- β110.Ong/ml group and PD98059 72J/cm2 TGF-β110.0ng / ml, respectively processing fibroblasts using methyl thiazolyl tetrazolium bromide (MTT) assay of cell proliferation; application Western blot ( western blot) assay groups phosphorylated extracellular signal-regulated kinase (p-ERK) changes. Vitro develop into fiber cells through I, collagen type III staining positive for cytokeratin-negative cells with fibroblast morphology. 2.1 fibroblasts treated with different concentrations of transforming growth factor-β1, in comparison with the control group, each group concentration of TGF-β1, 24 and 48 hours, respectively, 0.01 ng, 0.1 ng / ml, and 1.0ng/ml 50.0ng/ml of TGF-β1 on fibroblast proliferation activity there was no significant increase (P gt; 0.05), TGF-β1 concentration 10.0ng/ml cell proliferation activity than the control group was significantly higher (P lt; 0.05 ). 2.2, compared with the control group: TGF-β1 concentration group and time group p-ERK protein expression are different phosphorylated ERK protein expression levels gradually increased with the increase in the dose of TGF-β1 and prolonged duration of action, TGF-β1 10.0ng/ml p-ERK reached its peak in the 15th minute, phosphorylated ERK protein levels began to increase, p-ERK expression levels reached a peak in 60 minutes. 120 minutes when the expression of p-ERK attenuation. 2.3 The the inhibitor intervention group compared with the control group: the IPL irradiation and TGF-β1 for 24 and 48 hours after TGF-β110.0ng/ml the the IPL irradiation dose 72J/cm2 group 72J/cm2 of TGF-β110 .0ng/ml composed fibroblast proliferation activity was significantly increased (P lt; 0.01), phosphorylated ERK expression levels also increase. And PD98059 TGF-β110.0ng/ml, PD98059 72J/cm2 than control group cells proliferation activity decreased (P lt; 0.05), but a slight increase in the level of expression of phosphorylated ERK. I group showed no change. Conclusions large doses of TGF-β1 can promote the proliferation of fibroblasts cultured in vitro, and can increase intracellular the ERK signaling protein activity. P-ERK levels of TGF-β1-induced fibroblast may be dependent on the activation of the ERK pathway. Intense pulsed light can promote the proliferation of fibroblasts in vitro, its proliferation may be one mechanism through the ERK signaling pathway. 3. TGF-β1 and IPL synergistic interaction may exist in the proliferation of fibroblasts.