H2-RLX抑制SSc皮损成纤维细胞增殖和向肌成纤维细胞转分化的研究

Studies on the fibroblast proliferation and transdifferentiation for myofibroblast from skin lesion of the patients with systemic sclerosis repressed by H2 Relaxin

目的:探讨成纤维细胞(FB)向肌成纤维细胞(MFB)转分化在系统性硬化症(SSc)发病机制中的作用和H2松弛素(H2-RLX)在SSc中的抗纤维化作用机制。方法:体外培养SSc患者皮损和正常皮肤FB及鉴定;免疫细胞化学法定性、ELISA法定量检测FB中α-平滑肌肌动蛋白(α-SMA)而了解MFB比重;施加并观察H2-RLX对SSc FB增殖和转分化为MFB的影响。结果:两组FB的细胞形态无明显不同;SSc组α-SMA阳性率均值高于对照组(P<0.01);随培养时间的延长,两组α-SMA量均渐增多(P均<0.01),但在培养的24、48、72 h,SSc组α-SMA量分别高于对照组(P均<0.05);H2-RLX 1μg/L对FB增殖和α-SMA量无明显影响,而10μg/L和100μg/L则完全地抑制FB增殖和α-SMA量(P均<0.05),以100μg/L时抑制作用最强。结论:SSc患者皮损来源的FB存在强烈地向MFB转分化的特性,H2-RLX则可通过抑制FB增殖及转分化为MFB而在SSc中发挥抗纤维化作用。

AIM： The study was to explore the effects of the fibroblast transdifferentiation for myofibroblast （MFB） in the pathogenesis of systemic sclerosis （SSc） and to research the effect mechanism of H2 Relaxin （H2-RLX） against fibrosis in SSc. METHODS： The fibroblasts derived from the skin lesion of the SSc patients and normal skin tissue were respectively cultured in vitro and demonstrated. The MFB proportion in fibroblast was known by α-smooth muscle actin （a-SMA） in fibroblast culture detected with immunohistochemistry for qualitative study and ELISA for quantitative analysis. The influence on fibroblast proliferation and transdifferentiation for MFB was observed by adding H2- RLX. RESULTS： There were no apparent differences for cell morphology between the fibroblasts from SSc patients and controls. The means of positive α-SMA in SSc group were higher than those in controls （ P 〈 0. 01 ）. With culture time extended, α-SMA levels of the two groups all increased gradually （P〈0.01 all）, but there were higher α-SMA levels in SSc group than those in controls separately at H24, H48, H72 in culture （P 〈 0.05 all）. Fibroblast proliferation and α-SMA levels were not influenced after adding 1μg/L of H2-RLX, but 10 μg/L and 100 μg/L of it could completely in- hibit the fibroblast proliferation and α-SMA levels （ P 〈0. 05 all）, with the strongest repressing effect after adding 100 μg/L of it. CONCLUSION： There is a specific property of fibroblasts transdifferentiation for MFB strongly from the skin lesion of the SSc patients. H2-RLX could give play to the antifibrotic effects by repressing the fibroblast proliferation and transdifferentiation for MFB in SSc.