藏雪蓮多糖減輕UVB輻射后HaCaT細胞氧化損傷的實驗研究
發(fā)布時間:2018-06-30 00:16
本文選題:紫外線輻射 + 人永生化角質細胞。 參考:《青海大學》2017年碩士論文
【摘要】:目的研究藏雪蓮植物多糖是否能對體外UVB(Ultraviolet b,中波紫外線)誘導的人永生化角質形成細胞(Ha Ca T細胞)的氧化損傷起保護作用。方法應用體外培養(yǎng)的細胞實驗模型培養(yǎng)人永生化表皮(Ha Ca T細胞),按照輻射能量不同,分為高輻射組,低輻射組。應用提純后不同濃度的藏雪蓮多糖對輻射細胞進行干預,分為對照組(正常培養(yǎng),無輻射,無多糖)、輻射組(輻射,無多糖)、低濃度臧雪蓮多糖組、高濃度臧雪蓮多糖組。多糖提前1h添加。照射后各組細胞培養(yǎng)18h。應用臺盼藍染色法對輻射后各組細胞形態(tài)與凋亡狀況觀察并攝片,應用MTS法測細胞增殖;抗氧化酶測定:酶標法測GSH(谷胱甘肽)含量,TBA法測定MDA(細胞丙二醛)含量,可見光法測定CAT(過氧化氫酶)及WST-1法測定SOD(超氧化物歧化酶)活力。結果與輻射組相比,給予不同濃度的藏雪蓮多糖后,藍染數(shù)下降,細胞結構完整性上升,減少細胞的死亡率;增殖活性提高;降低MDA含量,使GSH含量、SOD及CAT活性上升,與輻射組比較均具有統(tǒng)計學差異(p0.05)。結論經(jīng)過不同能量UVB輻射后的Ha Ca T細胞在給予一定劑量的藏雪蓮多糖形后,可有效減輕光損傷造成的細胞結構破壞,減少細胞凋亡。
[Abstract]:Objective to study whether the polysaccharides of Saussurea lanceolata can protect human immortalized keratinocytes (Ha Ca T cells) from oxidative damage induced by UVB (Ultraviolet B) in vitro. Methods Human immortalized epidermis (Ha Ca T cells) were cultured in vitro and divided into high radiation group and low radiation group according to different radiation energy. After purification, different concentrations of Tibetan snow lotus polysaccharides were used to interfere with radiation cells and were divided into control group (normal culture, no radiation, no polysaccharide), radiation group (radiation, no polysaccharide), low concentration Zang Xuelian polysaccharide group and high concentration Zang Xuelian polysaccharide group. Polysaccharide was added 1 hour earlier. The cells were cultured for 18 hours after irradiation. Cell morphology and apoptosis were observed by trypan blue staining, cell proliferation was measured by MTS method, GSH (glutathione) content was measured by enzyme labeling method and MDA (cell malondialdehyde) content was measured by TBA method. Cat (catalase) and SOD (superoxide dismutase) activity were measured by visible light method and WST-1 method respectively. Results compared with radiation group, the blue staining number decreased, cell structural integrity increased, cell death rate decreased, proliferation activity increased, MDA content decreased, and GSH content SOD and cat activities increased, compared with radiation group. There was statistical difference between the two groups (p0.05). Conclusion the Ha-Ca T cells irradiated with different energy UVB can effectively reduce the damage of cell structure and apoptosis caused by light damage after given a certain dose of polysaccharide form of Saussurea sorbifolia.
【學位授予單位】:青海大學
【學位級別】:碩士
【學位授予年份】:2017
【分類號】:R751
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