柴达木枸杞多糖对UVB诱导人HaCaT细胞氧化损伤及凋亡相关蛋白的影响

发布时间：2018-09-07 22:07

【摘要】：目的观察枸杞多糖对中波紫外线(UVB)辐射后人角质形成(HaCaT)细胞的氧化损伤及p38丝裂原活化蛋白激酶(p38MAPK)、半胱氨酸蛋白酶-8(caspase-8)、半胱氨酸蛋白酶-3(caspase-3)、B细胞淋巴瘤因子2(Bcl-2)表达的影响。方法超声辅助水提法制备枸杞多糖(LBP)粉末,体外培养HaCaT细胞,随机分为空白对照组、UVB照射组(UVB 30m J/cm~2辐射40min)、UVB+LBP组(UVB 30m J/cm~2辐射40min+LBP 1mg/m L),照射前12h加入LBP,照射结束后继续培养20h,倒置显微镜观察细胞形态;MTS法检测各组细胞吸光度值(A值);酶生化法检测超氧化物歧化酶(SOD)活力、过氧化氢酶(CAT)活力和谷胱甘肽过氧化物酶(GSH-Px)含量和丙二醛(MDA)含量;Western blot法测定各组细胞p38MAPK,caspase-8,caspase-3和Bcl-2的表达水平。结果与对照组相比,UVB照射组细胞形态明显异常,漂浮的死细胞明显增多;与UVB照射组相比,UVB+LBP组细胞的形态趋于正常,漂浮的死细胞明显减少。UVB照射组较对照组细胞吸光度(A值)明显降低(P0.05);UVB+LBP组细胞吸光度较UVB照射组明显升高(P0.05)。UVB照射组SOD活力、CAT活力、GSH-Px含量较对照组降低,MDA含量较对照组升高(P0.05);UVB+LBP组细胞SOD活力、CAT活力、GSH-Px含量较UVB照射组升高,MDA含量较UVB照射组降低(P0.05)。UVB照射组p38MAPK,caspase-8,caspase-3蛋白表达量明显高于对照组,Bcl-2表达量明显低于对照组(P0.05);与UVB照射组相比,UVB+LBP组HaCaT细胞p38MAPK,caspase-8,caspase-3表达水平降低,Bcl-2表达水平升高(P0.05)。结论枸杞多糖可抑制UVB辐射后HaCaT细胞的氧化损伤,并下调p38MAPK,caspase-8,caspase-3及上调Bcl-2的表达水平。
[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.
【作者单位】：青海大学附属医院皮肤科;
【基金】：青海省科学计划应用基础研究项目(2014-ZJ-756)
【分类号】：R751

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