【摘要】：目的觀察枸杞多糖對(duì)中波紫外線(UVB)輻射后人角質(zhì)形成(HaCaT)細(xì)胞的氧化損傷及p38絲裂原活化蛋白激酶(p38MAPK)、半胱氨酸蛋白酶-8(caspase-8)、半胱氨酸蛋白酶-3(caspase-3)、B細(xì)胞淋巴瘤因子2(Bcl-2)表達(dá)的影響。方法超聲輔助水提法制備枸杞多糖(LBP)粉末,體外培養(yǎng)HaCaT細(xì)胞,隨機(jī)分為空白對(duì)照組、UVB照射組(UVB 30m J/cm~2輻射40min)、UVB+LBP組(UVB 30m J/cm~2輻射40min+LBP 1mg/m L),照射前12h加入LBP,照射結(jié)束后繼續(xù)培養(yǎng)20h,倒置顯微鏡觀察細(xì)胞形態(tài);MTS法檢測(cè)各組細(xì)胞吸光度值(A值);酶生化法檢測(cè)超氧化物歧化酶(SOD)活力、過氧化氫酶(CAT)活力和谷胱甘肽過氧化物酶(GSH-Px)含量和丙二醛(MDA)含量;Western blot法測(cè)定各組細(xì)胞p38MAPK,caspase-8,caspase-3和Bcl-2的表達(dá)水平。結(jié)果與對(duì)照組相比,UVB照射組細(xì)胞形態(tài)明顯異常,漂浮的死細(xì)胞明顯增多;與UVB照射組相比,UVB+LBP組細(xì)胞的形態(tài)趨于正常,漂浮的死細(xì)胞明顯減少。UVB照射組較對(duì)照組細(xì)胞吸光度(A值)明顯降低(P0.05);UVB+LBP組細(xì)胞吸光度較UVB照射組明顯升高(P0.05)。UVB照射組SOD活力、CAT活力、GSH-Px含量較對(duì)照組降低,MDA含量較對(duì)照組升高(P0.05);UVB+LBP組細(xì)胞SOD活力、CAT活力、GSH-Px含量較UVB照射組升高,MDA含量較UVB照射組降低(P0.05)。UVB照射組p38MAPK,caspase-8,caspase-3蛋白表達(dá)量明顯高于對(duì)照組,Bcl-2表達(dá)量明顯低于對(duì)照組(P0.05);與UVB照射組相比,UVB+LBP組HaCaT細(xì)胞p38MAPK,caspase-8,caspase-3表達(dá)水平降低,Bcl-2表達(dá)水平升高(P0.05)。結(jié)論枸杞多糖可抑制UVB輻射后HaCaT細(xì)胞的氧化損傷,并下調(diào)p38MAPK,caspase-8,caspase-3及上調(diào)Bcl-2的表達(dá)水平。
[Abstract]:Objective to investigate the effects of Lycium barbarum polysaccharides on oxidative damage and expression of p38 mitogen-activated protein kinase (p38MAPK), cysteine protease-8 (caspase-8) and cysteine protease -3 (caspase-3) / B cell lymphoma factor 2 (Bcl-2) in human keratinocytes induced by ultraviolet (UVB) radiation. Methods Ultrasound-assisted extraction of Lycium barbarum polysaccharides (LBP) powder was prepared and HaCaT cells were cultured in vitro. They were randomly divided into the blank control group (UVB 30m J/cm~2 radiation 40min) and the UVB LBP group (UVB 30m J/cm~2 radiation 40min LBP 1mg/m L), irradiation 12h before 40min LBP 1mg/m L), irradiation) and cultured for 20h after LBP, irradiation. The cell morphology was observed by inverted microscope to detect the absorbance value of cells in each group. The activity of superoxide dismutase (SOD) was detected by enzyme biochemical method. Catalase (CAT) activity, glutathione peroxidase (GSH-Px) content and malondialdehyde (MDA) (MDA) content were determined by Western blot method. Results compared with the control group, the morphology of the cells in the UVB irradiation group was obviously abnormal, and the number of dead cells floating in the UVB irradiation group was obviously increased, and the morphology of the cells in the UVB LBP group was normal compared with that in the UVB irradiation group. The cell absorbance in UVB LBP group was significantly higher than that in UVB irradiation group (P0.05). GSH-Px content of SOD activity cat activity in UVB irradiation group was lower than that in control group. The level of SOD activity cat activity and GSH-Px in UVB LBP group was significantly lower than that in UVB irradiation group (P0.05). The expression of p38 MAPKCaspase-8 caspase-8 caspase-3 protein in UVB irradiation group was significantly higher than that in control group (P0.05), and that in UVB irradiation group was significantly lower than that in control group (P0.05). Compared with UVB LBP group, the expression level of p38 MAPKT caspase-8 and caspase-3 in HaCaT cells was significantly lower than that in UVB LBP group (P0.05). Conclusion Lycium barbarum polysaccharides can inhibit oxidative damage of HaCaT cells after UVB irradiation and down-regulate the expression of p38 MAPK- caspase-8 caspase-3 and up-regulate the expression of Bcl-2.
【作者單位】： 青海大學(xué)附屬醫(yī)院皮膚科;
【基金】：青海省科學(xué)計(jì)劃應(yīng)用基礎(chǔ)研究項(xiàng)目(2014-ZJ-756)
【分類號(hào)】：R751

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