【摘要】 目的探讨蜕皮甾酮(ecdysterone,EDS)对中波紫外线(UVB)诱导角质形成细胞损伤的保护作用及机制。方法将培养的HaCaT细胞分为正常对照组、UVB组、2.0μmol/L蜕皮甾酮剂量组(UVB+2.0μmol/L蜕皮甾酮),1.5μmol/L蜕皮甾酮剂量组(UVB+1.5μmol/L蜕皮甾酮),1.0μmol/L蜕皮甾酮剂量组(UVB+1.0μmol/L蜕皮甾酮);CCK-8法检测蜕皮甾酮对HaCaT细胞增殖能力的影响;Hoechst33258荧光染色法观察细胞凋亡形态;采用PI单染和Annexin V-FITC/PI染色法,流式细胞仪检测HaCaT细胞凋亡率;比色法检测超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性及丙二醛(MDA)水平。Western印迹检测MMP-1,MMP-9,TIMP-1蛋白表达水平变化。结果在所选浓度范围内,蜕皮甾酮对HaCaT细胞的增殖无明显影响(P>0.05)。与正常对照组比较,UVB组HaCaT细胞出现明显的凋亡形态,凋亡率明显增高;1.0μmol/L、1.5μmol/L、2.0μmol/L蜕皮甾酮剂量组较UVB组凋亡细胞逐渐减少,差异有统计学意义(均P<0.01)。与正常对照组相比,UVB组HaCaT细胞SOD和GSH-Px活性降低,MDA含量升高(P<0.01);1.0μmol/L、1.5μmol/L、2.0μmol/L蜕皮甾酮剂量组与UVB组比较,SOD和GSH-Px活性增高,MDA含量下降(P<0.01)。Western印迹显示:UVB组HaCaT细胞MMP-l,MMP-9蛋白表达量明显高于正常对照组(P<0.01),而TIMP-l蛋白表达量低于正常对照组(P<0.01);1.0μmol/L、1.5μmol/L、2.0μmol/L蜕皮甾酮剂量组MMP-l,MMP-9表达量明显低于UVB组(P<0.01),而TIMP-l表达量明显高于UVB组(P<0.01)。结论蜕皮甾酮对中波紫外线诱导的HaCaT细胞凋亡,氧化损伤和光老化均具有一定的保护作用。更多还原

【Abstract】 Objective To investigate the effect of ecdysterone on HaCaT cells induced by UVB radiation. Methods HaCaT cells were cultured in vitro and divided into control group,2. 0 μmol/L EDS group( UVB + 2. 0μmol/L EDS),1. 5 μmol/L EDS group( UVB + 1. 5 μmol/L EDS),1. 0 μmol/L EDS group( UVB + 1. 0μmol/L EDS),and UVB group. The viability of HaCaT cells was detected by cell counting Kit-8 assay; The apoptosis of HaCaT cells was determined by flow cytometry assay and was observed by fluorescent staining.Colorimetry was performed to evaluate superoxide dismutase( SOD) and glutathione peroxidase( GSH-Px)activities and to determine malondialdehvde( MDA) levels. Western blotting was used to detect MMP-1,MMP-9,TIMP-1 protein expressions in HaCaT cells. Results Ecdysterone in selective concentration had no effect on the viability of HaCaT cells( P > 0. 05). Compared with control group,the levels of MDA and apoptosis rates of HaCaT cells were significantly increased and the levels of SOD,GSH-Px of HaCaT cells were significantly decreased in UVB group( P < 0. 01),but the levels of MDA and apoptosis rates of HaCaT cells injured by UVB radiation were decreased and the levels of SOD,GSH-Px of HaCaT cells were elevated after ecdysterone treatment in a dose-dependent manner( P < 0. 01). The protein expressions of MMP-1 and MMP-9 in HaCaT cells in the UVB group showed significant elevation in comparison with the control group,the expression of TIMP-1 protein was declined in the UVB group compared with the control group( P <0. 01),but 2. 0 μmol/L,1. 5 μmol/L,1. 0 μmol/L ecdysterone treatments down-regulated the protein expressions of MMP-1,MMP-9 and up-regulated the expression of TIMP-1 in UVB-radiated HaCaT cells( P <0. 01). Conclusion Ecdysterone has a protective effect on oxidative stress,apoptosis and skin photoaging induced by UVB radiation.更多还原