柴達木黑果枸杞多糖對UVB誘導(dǎo)HaCaT細胞的光損傷的保護作用
發(fā)布時間:2018-03-19 16:17
本文選題:柴達木黑果枸杞 切入點:枸杞多糖 出處:《青海大學(xué)》2017年碩士論文 論文類型:學(xué)位論文
【摘要】:目的:探討柴達木黑果枸杞(Lycium ruthenicum Murr.)多糖對中波紫外線(UVB)誘導(dǎo)HaCaT細胞光老化的防治作用及作用機制。方法:使用超聲輔助水提法提取柴達木黑果枸杞中的多糖成分,HaCaT細胞分為對照組、照射組、實驗組,實驗組將黑果枸杞多糖于UVB輻射前6 h加入細胞培養(yǎng)基中,各組照射后12 h,觀察細胞形態(tài),收集細胞MTS法測定細胞增殖,流式細胞儀測定細胞凋亡率。酶標法檢測細胞丙二醛(MDA)含量、超氧化物歧化酶(SOD)活力,谷胱甘肽(GSH-Px)含量,過氧化氫酶(CAT)活力。ELASI法測白介素1(IL-1)、腫瘤壞死因子a(TNF-a)。蛋白免疫印跡(Western Blotting)法檢測P16蛋白的表達。結(jié)果:與對照組細胞相比,UVB組細胞形態(tài)模糊不清,出現(xiàn)死亡漂浮現(xiàn)象,實驗組細胞腫脹,但形態(tài)尚清楚。UVB組細胞增殖率及細胞凋亡率下,MDA含量明顯上升,SOD活性、GSH含量、CAT活性明顯下降,上清中IL-1和TNF-α含量及細胞中TNF-α含量均明顯升高,P16蛋白表達升高,差異具有統(tǒng)計學(xué)意義(均P0.05)。與UVB組比較,實驗組細胞增殖率及細胞凋亡率增加,MDA含量明顯降低,SOD活性、GSH含量、CAT活性明顯升高,上清及細胞中TNF-α含量明顯下降,細胞中IL-1升高,差異具有統(tǒng)計學(xué)意義(P0.05)。結(jié)論:柴達木黑果枸杞多糖對HaCaT細胞的光損傷有保護作用,其保護機制可能與減少細胞炎性物質(zhì)的合成和分泌、減少自由基及降低P16蛋白表達有關(guān)。
[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- 偽) and tumor necrosis factor (TNF- 偽). 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- 偽 in the supernatant and the content of TNF- 偽 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- 偽 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.
【學(xué)位授予單位】:青海大學(xué)
【學(xué)位級別】:碩士
【學(xué)位授予年份】:2017
【分類號】:R751
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1 郭音;有氧運動與黑果枸杞多糖改善小鼠阻塞性黃疸致肝損傷的研究[D];湖南師范大學(xué);2016年
2 任立汆;柴達木黑果枸杞多糖對UVB誘導(dǎo)HaCaT細胞的光損傷的保護作用[D];青海大學(xué);2017年
3 呂曉鵬;黑果枸杞多糖的分離純化與結(jié)構(gòu)表征研究[D];西北大學(xué);2012年
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