The Effect of HMGB-1on Skin Fibroblasts Activation and the Expression of Col Ⅰ, Fn in SSc
|School||Central South University|
|Keywords||HMGB-1 systemic sclerosis skin fibroblasts α-smoothmuscle actin collagen type one Ⅰ fibronectin|
Objective:To explore the effect of High Mobility Group Box Chromosomal Protein1(HMGB-1) on health human and systemic sclerosis (SSc) skin fibroblasts activation and the expression of Col I,Fn in vivo.Methods:Fibroblasts derived from the normal skin tissue and skin lesions of SSc patients were cultured in vivo, identified by immunocytochemistry and morphology, select4-6cells for experiment. HMGB1stimulates skin fibroblasts for24,48and72h in different final concentrations, MTT assay was used to observe the proliferation of skin fibroblasts; HMGB1stimulates skin fibroblasts for48h in a final concentration of0.5μg/ml, the concentration of the control group is0μg/ml, Western-Blot assay (WB) was used to observe the expression of the a-smooth muscle actin (α-SMA) protein; Real-Time PCR method was used to detect the expression of the collagen type one Ⅰ (Col Ⅰ) and fibronectin (Fn) mRNA.Results:1.The two groups of cells isolated from SSc patients and control subjects showed a similar morphology, and both were identified as fibroblasts by immunicytochemistry;2. After24,48and72h stimulates in different concentrations, MTT assay results suggest that HMGB-1slightly inhibits normal human skin fibroblast proliferation in concentration of0.5μg/ml, and has no effect on the fibroblast of the patients with SSc;3. WB show that the expression of α-SMA of normal and SSc skin fibroblasts are significantly higher than the control group (p <0.05) after48h HMGB1stimulating, and the expression of SSc skin fibroblasts is significantly higher than the normal group(p<0.01); Real-Time PCR results that the mRNA expression of Col I and Fn of SSc skin fibroblasts is significantly higher than control and normal skin fibroblasts after48h HMGB1stimulating, and the expression in the normal group is indifferent whether stimulating or not.Conclusion:1. HMGB1can induce normal and SSc skin fibroblasts transform into myofibroblast;2. HMGB1can induce SSc skin fibroblasts synthetizing extracellular matrix, suggesting that HMGB1may be involved in the pathological process of skin fibrosis in SSc.