[1]胡惠清,刘斌,李静.蜕皮甾酮对UVB诱导HaCaT细胞损伤的保护作用[J].中国皮肤性病学杂志,2017,(07):718-722.[doi:10.13735/j.cjdv.1001-7089.201702025]
 HU Hui-qing,LIU Bin,LI Jing.Protective Effect of Ecdysterone Against Ultraviolet B-induced Damage to HaCaT Cells[J].The Chinese Journal of Dermatovenereology,2017,(07):718-722.[doi:10.13735/j.cjdv.1001-7089.201702025]
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蜕皮甾酮对UVB诱导HaCaT细胞损伤的保护作用
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《中国皮肤性病学杂志》[ISSN:1001-7089/CN:61-1197/R]

卷:
期数:
2017年07期
页码:
718-722
栏目:
论著
出版日期:
2017-07-01

文章信息/Info

Title:
Protective Effect of Ecdysterone Against Ultraviolet B-induced Damage to HaCaT Cells
作者:
胡惠清刘斌李静
武汉市第九医院皮肤科,湖北 武汉 438000
Author(s):
HU Hui-qingLIU BinLI Jing
Department of dermatology Ninth hospital of wuhan, Wuhan 438000, China
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
Keywords:
HaCaT cells ultraviolet B ecdysterone Matrixmetalloproteinases-1 Matrixmetalloproteinases-9 Tissue inhibitor of metalloproteinases-1
分类号:
R 758.1
DOI:
10.13735/j.cjdv.1001-7089.201702025
文献标志码:
A
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、2.0 μmol/L蜕皮甾酮剂量组(UVB + 2.0 μmol/L 蜕皮甾酮),1.5 μmol/L蜕皮甾酮剂量组(UVB+1.5 μmol/L蜕皮甾酮),1.0 μmol/L蜕皮甾酮剂量组(UVB+1.0 μmol/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; Hoechst 33258 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; 1.0 μmol/L、1.5 μmol/L、2.0 μmol/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); 1.0 μmol/L、1.5 μmol/L、2.0 μmol/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); 1.0 μmol/L、1.5 μmol/L、2.0 μmol/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。
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+2.0 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.

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更新日期/Last Update: 2017-06-30