蜕皮甾酮对UVB诱导HaCaT细胞损伤的保护作用

发布时间：2018-03-03 05:34

  本文选题：HaCaT细胞　切入点：中波紫外线　出处：《中国皮肤性病学杂志》2017年07期 　论文类型：期刊论文

【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(P0.05)。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P0.01)。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(P0.01);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(P0.01)。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(P0.01),而TIMP-l蛋白表达量低于正常对照组(P0.01);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(P0.01),而TIMP-l表达量明显高于UVB组(P0.01)。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。
[Abstract]:Objective to investigate the protective effect and mechanism of ecdysterone ecdysterone (EDS) on keratinocyte damage induced by ultraviolet B (UVB). Methods cultured HaCaT cells were divided into two groups: UVB 2.0 渭 mol/L ecdysterone group and 1.5 渭 mol/L ecdysterone group. The effect of UVB 1.5 渭 mol/L ecdysterone and 1.0 渭 mol/L ecdysterone on the proliferation of HaCaT cells was detected by CCK-8 method. The morphology of apoptosis was observed by Hoechst33258 fluorescence staining. Pi single staining and Annexin V-FITC / Pi staining were used to detect the apoptosis rate of HaCaT cells. The activity of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and the level of malondialdehyde (MDAs) were detected by colorimetric assay. Western blot was used to detect the expression of MMP-1, MMP-9 and TIMP-1. There was no obvious effect of ecdysterone on the proliferation of HaCaT cells. Compared with the normal control group, the apoptosis rate of HaCaT cells in UVB group was significantly higher than that in UVB group, and the apoptosis rate was significantly increased. The apoptosis rate in the group of 1.0 渭 mol 路L ~ (-1) L ~ (-1) 路L ~ (-1) 渭 mol 路L ~ (-1) and 2.0 渭 mol / L _ (2) 渭 mol/L ecdysterone decreased gradually compared with that in the group of UVB. Compared with the normal control group, the activity of SOD and GSH-Px in HaCaT cells decreased significantly (P 0.01). The activity of SOD and GSH-Px in UVB cells decreased. The activity of mol/L and GSH-Px in UVB group was decreased compared with that in UVB group. Western blotting showed that HaCaT was fine in the group of GSH-Px in the group of 1. 01 渭 mol 路L ~ (-1) 渭 mol 路L ~ (-1) 路L ~ (-1) 路L ~ (-1) 路L ~ (-1) 路L ~ (-1), compared with that in the group of UVB (P ~ (0.01) / L ~ (-1)). The expression of MMP-lMP-9 protein was significantly higher than that of the normal control group (P0.01A), while the expression of TIMP-l protein was lower than that of the control group (P0.01mol / L = 1.0 渭 mol / L ~ (1.5) 渭 mol / L ~ (1.5) 渭 mol / L ~ (2) 渭 mol 路L ~ (-1)) significantly lower than that of the UVB group (P _ (0.01)), and the expression of TIMP-l was significantly higher than that of the UVB group (P _ (0.01)). Conclusion the expression of MMP _ 9 is significantly higher than that of the UVB group (P _ (0.01)). Ultraviolet B induced apoptosis of HaCaT cells. Both oxidative damage and photoaging have some protective effects.
【作者单位】： 武汉市第九医院皮肤科;
【分类号】：R758.1
，

本文编号：1559758