本文選題：HaCaT細(xì)胞　切入點(diǎn)：中波紫外線　出處：《中國(guó)皮膚性病學(xué)雜志》2017年07期 　論文類(lèi)型：期刊論文

【摘要】：目的探討蛻皮甾酮(ecdysterone,EDS)對(duì)中波紫外線(UVB)誘導(dǎo)角質(zhì)形成細(xì)胞損傷的保護(hù)作用及機(jī)制。方法將培養(yǎng)的HaCaT細(xì)胞分為正常對(duì)照組、UVB組、2.0μmol/L蛻皮甾酮?jiǎng)┝拷M(UVB+2.0μmol/L蛻皮甾酮),1.5μmol/L蛻皮甾酮?jiǎng)┝拷M(UVB+1.5μmol/L蛻皮甾酮),1.0μmol/L蛻皮甾酮?jiǎng)┝拷M(UVB+1.0μmol/L蛻皮甾酮);CCK-8法檢測(cè)蛻皮甾酮對(duì)HaCaT細(xì)胞增殖能力的影響;Hoechst33258熒光染色法觀察細(xì)胞凋亡形態(tài);采用PI單染和Annexin V-FITC/PI染色法,流式細(xì)胞儀檢測(cè)HaCaT細(xì)胞凋亡率;比色法檢測(cè)超氧化物歧化酶(SOD)、谷胱甘肽過(guò)氧化物酶(GSH-Px)活性及丙二醛(MDA)水平。Western印跡檢測(cè)MMP-1,MMP-9,TIMP-1蛋白表達(dá)水平變化。結(jié)果在所選濃度范圍內(nèi),蛻皮甾酮對(duì)HaCaT細(xì)胞的增殖無(wú)明顯影響(P0.05)。與正常對(duì)照組比較,UVB組HaCaT細(xì)胞出現(xiàn)明顯的凋亡形態(tài),凋亡率明顯增高;1.0μmol/L、1.5μmol/L、2.0μmol/L蛻皮甾酮?jiǎng)┝拷M較UVB組凋亡細(xì)胞逐漸減少,差異有統(tǒng)計(jì)學(xué)意義(均P0.01)。與正常對(duì)照組相比,UVB組HaCaT細(xì)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蛻皮甾酮?jiǎng)┝拷M與UVB組比較,SOD和GSH-Px活性增高,MDA含量下降(P0.01)。Western印跡顯示:UVB組HaCaT細(xì)胞MMP-l,MMP-9蛋白表達(dá)量明顯高于正常對(duì)照組(P0.01),而TIMP-l蛋白表達(dá)量低于正常對(duì)照組(P0.01);1.0μmol/L、1.5μmol/L、2.0μmol/L蛻皮甾酮?jiǎng)┝拷MMMP-l,MMP-9表達(dá)量明顯低于UVB組(P0.01),而TIMP-l表達(dá)量明顯高于UVB組(P0.01)。結(jié)論蛻皮甾酮對(duì)中波紫外線誘導(dǎo)的HaCaT細(xì)胞凋亡,氧化損傷和光老化均具有一定的保護(hù)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.
【作者單位】： 武漢市第九醫(yī)院皮膚科;
【分類(lèi)號(hào)】：R758.1
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本文編號(hào)：1559758