柴达木黑果枸杞多糖对UVB诱导HaCaT细胞的光损伤的保护作用

发布时间：2018-03-19 16:17

本文选题：柴达木黑果枸杞　切入点：枸杞多糖　出处：《青海大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:探讨柴达木黑果枸杞(Lycium ruthenicum Murr.)多糖对中波紫外线(UVB)诱导HaCaT细胞光老化的防治作用及作用机制。方法:使用超声辅助水提法提取柴达木黑果枸杞中的多糖成分,HaCaT细胞分为对照组、照射组、实验组,实验组将黑果枸杞多糖于UVB辐射前6 h加入细胞培养基中,各组照射后12 h,观察细胞形态,收集细胞MTS法测定细胞增殖,流式细胞仪测定细胞凋亡率。酶标法检测细胞丙二醛(MDA)含量、超氧化物歧化酶(SOD)活力,谷胱甘肽(GSH-Px)含量,过氧化氢酶(CAT)活力。ELASI法测白介素1(IL-1)、肿瘤坏死因子a(TNF-a)。蛋白免疫印迹(Western Blotting)法检测P16蛋白的表达。结果:与对照组细胞相比,UVB组细胞形态模糊不清,出现死亡漂浮现象,实验组细胞肿胀,但形态尚清楚。UVB组细胞增殖率及细胞凋亡率下,MDA含量明显上升,SOD活性、GSH含量、CAT活性明显下降,上清中IL-1和TNF-α含量及细胞中TNF-α含量均明显升高,P16蛋白表达升高,差异具有统计学意义(均P0.05)。与UVB组比较,实验组细胞增殖率及细胞凋亡率增加,MDA含量明显降低,SOD活性、GSH含量、CAT活性明显升高,上清及细胞中TNF-α含量明显下降,细胞中IL-1升高,差异具有统计学意义(P0.05)。结论:柴达木黑果枸杞多糖对HaCaT细胞的光损伤有保护作用,其保护机制可能与减少细胞炎性物质的合成和分泌、减少自由基及降低P16蛋白表达有关。
[Abstract]:Objective: to investigate the effect of Lycium ruthenicum Murr.) polysaccharides on photoaging of HaCaT cells induced by UVB. Methods: extraction of polysaccharides from Lycium ruthenicum Murr. by ultrasonic assisted extraction. HaCaT cells were divided into control group. In the irradiation group, experimental group and experimental group, the polysaccharides of Lycium barbarum were added to the cell culture medium 6 hours before UVB irradiation. After 12 hours of irradiation, cell morphology was observed and cell proliferation was measured by MTS method. The content of malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione (GSH-Px) were measured by flow cytometry. The activity of catalase CAT.ELASI method was used to detect the expression of P16 protein in UVB group, and the expression of P16 protein was detected by Western blotting.Results: compared with the control group, the expression of P16 protein was detected by ELASI method, tumor necrosis factor (TNF- 伪) and tumor necrosis factor (TNF- 伪). Results: compared with the control group, the cell morphology of UVB group was blurred, and death floated. The cells in the experimental group were swollen, but the cell proliferation rate and apoptosis rate in the UVB group were obviously increased. The activity of IL-1 and TNF- 伪 in the supernatant and the content of TNF- 伪 in the supernatant were all significantly increased, and the expression of P16 protein was significantly increased in the supernatant. The difference was statistically significant (P 0.05). Compared with the UVB group, the cell proliferation rate and apoptosis rate in the experimental group increased the content of MDA, the activity of catalase, the content of GSH and cat in the supernatant, the content of TNF- 伪 in the supernatant, and the increase of IL-1 in the cell. Conclusion: Chaidamu Lycium barbarum polysaccharide has protective effect on light damage of HaCaT cells, and its protective mechanism may be related to reducing the synthesis and secretion of inflammatory substances, reducing free radicals and P16 protein expression.
【学位授予单位】：青海大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R751

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