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  本文選題：HaCaT細胞　切入點：中波紫外線　出處：《中國皮膚性病學(xué)雜志》2017年07期 　論文類型：期刊論文

【摘要】：目的探討蛻皮甾酮(ecdysterone,EDS)對中波紫外線(UVB)誘導(dǎo)角質(zhì)形成細胞損傷的保護作用及機制。方法將培養(yǎng)的HaCaT細胞分為正常對照組、UVB組、2.0μmol/L蛻皮甾酮劑量組(UVB+2.0μmol/L蛻皮甾酮),1.5μmol/L蛻皮甾酮劑量組(UVB+1.5μmol/L蛻皮甾酮),1.0μmol/L蛻皮甾酮劑量組(UVB+1.0μmol/L蛻皮甾酮);CCK-8法檢測蛻皮甾酮對HaCaT細胞增殖能力的影響;Hoechst33258熒光染色法觀察細胞凋亡形態(tài);采用PI單染和Annexin V-FITC/PI染色法,流式細胞儀檢測HaCaT細胞凋亡率;比色法檢測超氧化物歧化酶(SOD)、谷胱甘肽過氧化物酶(GSH-Px)活性及丙二醛(MDA)水平。Western印跡檢測MMP-1,MMP-9,TIMP-1蛋白表達水平變化。結(jié)果在所選濃度范圍內(nèi),蛻皮甾酮對HaCaT細胞的增殖無明顯影響(P0.05)。與正常對照組比較,UVB組HaCaT細胞出現(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蛻皮甾酮劑量組較UVB組凋亡細胞逐漸減少,差異有統(tǒng)計學(xué)意義(均P0.01)。與正常對照組相比,UVB組HaCaT細胞SOD和GSH-Px活性降低,MDA含量升高(P0.01);1.0μmol/L、1.5μmol/L、2.0μmol/L蛻皮甾酮劑量組與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細胞MMP-l,MMP-9蛋白表達量明顯高于正常對照組(P0.01),而TIMP-l蛋白表達量低于正常對照組(P0.01);1.0μmol/L、1.5μmol/L、2.0μmol/L蛻皮甾酮劑量組MMP-l,MMP-9表達量明顯低于UVB組(P0.01),而TIMP-l表達量明顯高于UVB組(P0.01)。結(jié)論蛻皮甾酮對中波紫外線誘導(dǎo)的HaCaT細胞凋亡,氧化損傷和光老化均具有一定的保護作用。
[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;
【分類號】：R758.1
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本文編號：1559758