蜕皮甾酮对UVB诱导HaCaT细胞损伤的保护作用  

Protective Effect of Ecdysterone Against Ultraviolet B-induced Damage to Ha Ca T Cells

作　　者：胡惠清 刘斌 李静 

机构地区：[1]武汉市第九医院皮肤科,湖北武汉438000

出　　处：《中国皮肤性病学杂志》2017年第7期718-722,共5页The Chinese Journal of Dermatovenereology

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。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＋2.0 μmol/L EDS）,1.5 μmol/L EDS group（UVB＋1.5 μmol/L EDS）, 1.0 μmol/L EDS group（UVB＋1.0 μmol/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.

关 键 词：HacaT细胞 中波紫外线 蜕皮甾酮 基质金属蛋白酶1 基质金属蛋白酶9 基质金属蛋白酶抑制因子1 

分 类 号：R758.1[医药卫生—皮肤病学与性病学]