The Expression of Cortistatin in Psoriasis and the Effect of Exogenous Cortistation on a Proliferation Model of EGF-induced HaCaT Cell
|School||Central South University|
|Course||Skin and Venereology|
|Keywords||HaCaT cells Cortex endostatin Ki-67 EGF Proliferation Psoriasis|
The purpose of detection of cortical endostatin expression in plasma and skin of patients with psoriasis vulgaris (CST), and the application of exogenous CST HaCaT cell proliferation observed in HaCaT cell proliferation induced by epidermal growth factor (EGF) model, discussed the role of the CST in the pathogenesis of psoriasis. The method 1.ELISA assay CST expression in the plasma of 72 patients with psoriasis vulgaris and 76 cases of normal control group, immunohistochemical method detected 14 cases of psoriasis patients (from the 72 cases) lesional and normal CST's expression in control skin. 2.1 × 106 / holes HaCaT cells were seeded in six-well plates and cultured for 24 hours, the cells immunohistochemical methods to detect the HaCaT CST protein expression, RT-PCR was used to detect the expression of CST mRNA. 5000 / hole inoculated HaCaT cells to 96-well plates, RPMI-1640 containing 10 ng / ml EGF Peiji When the cells adherent for the exogenous administration of different concentrations of CST (10-9-10-6M the) processing containing 10 ng / ml EGF HaCaT cells 24 hours, MTT assay was used to detect the change of cell proliferation, and repeated three times to observe the impact of exogenous CST EGF-induced HaCaT cell proliferation and select the best cortex endostatin concentration for subsequent experiments . 4 cell immunohistochemical methods to detect the CST and the EGF the HaCaT cell group (control group), containing 10 ng / ml EGF HaCaT cells (control group) and containing 10-6M CST and 10 ng / ml EGF HaCaT cells three groups (experimental group) of Ki-67 proliferation index changes. 5 enzyme-linked immunosorbent assay (ELISA) to detect changes in the three groups of second messenger cAMP. Results 1. The psoriasis patient plasma and skin lesions in the cortex endostatin expression than the normal control group were significantly lower (P lt; 0.05) but the expression levels of the psoriasis patient plasma CST PASI score no significant correlation (P gt; 0.05). The 2.CST protein in HaCaT cytoplasmic expression. And CST mRNA (173 bp) expression in HaCaT cells. 3 different concentrations CST (10-9-10-6M) in a concentration-dependent inhibition of 10 ng / ml EGF-induced proliferation of HaCaT cells. 10-6M best inhibitory effect, the inhibition rate of 54.62% (P values ??lt; 0.05) 4.Ki-67 expression in HaCaT cells of 10 ng / ml EGF treatment group (control group) increased more blank group 10-6M CST treatment, Ki-67 were down-regulated (P = lt; 0.05). 5 second messenger cAMP and Ki-67 change (P = lt; 0.05). Conclusion (1) patients with psoriasis vulgaris lesions and plasma CST than the normal control group decreased expression may be involved in the pathogenesis of psoriasis. (2) exogenous the CST can inhibit EGF-induced proliferation of HaCaT cells.