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目的:观察唐古特大黄多糖(Rheum tanguticum polysaccharide,RTP)对过氧化氢所致正常人肠上皮细胞损伤的保护作用。方法:以过氧化氢(100μmol/L)诱导正常人肠上皮细胞损伤,损伤前用RTP(30,100,300μg/ml)预处理细胞。采用MTT比色法测定细胞活力并进行形态学观察;吖啶橙染色法及流式细胞术观察细胞凋亡情况,采用分光光度法,用相应试剂盒测定乳酸脱氢酶(LDH)活性、丙二醛(MDA)含量及超氧化物歧化酶(SOD)活性。结果:加入过氧化氢后,细胞活力及SOD活性均降低,而MDA含量、LDH释放量及细胞凋亡数量均增高,与正常组比较具显著性差异(P<0.05或0.01);以RTP预处理细胞后发现,细胞活力明显增高,且SOD活性增强,同时,MDA水平、LDH活性均降低并可使凋亡细胞数减少。结论:RTP对过氧化氢所致的肠上皮细胞损伤具保护作用,且能抑制细胞凋亡及坏死,表明其具有一定的抗氧化损伤作用。
Objective: To observe the protective effect of Rheum tanguticum polysaccharide (RTP) on normal human intestinal epithelial cells induced by hydrogen peroxide. METHODS: Normal human intestinal epithelial cells were induced with hydrogen peroxide (100 μmol/L). Cells were pretreated with RTP (30, 100, 300 μg/ml) before injury. MTT colorimetry was used to determine cell viability and morphological observation. Acridine orange staining and flow cytometry were used to observe cell apoptosis. Spectrophotometry was used to determine lactate dehydrogenase (LDH) activity using the kit. Dialdehyde (MDA) content and superoxide dismutase (SOD) activity. RESULTS: After the addition of hydrogen peroxide, the cell viability and SOD activity were decreased, but the content of MDA, LDH release, and apoptosis were all increased, which was significantly different from the normal group (P<0.05 or 0.01); After treatment of the cells, it was found that cell viability was significantly increased and SOD activity was enhanced. At the same time, both MDA level and LDH activity were decreased and the number of apoptotic cells was decreased. Conclusion: RTP has a protective effect on hydrogen peroxide-induced intestinal epithelial cell damage, and can inhibit apoptosis and necrosis, indicating that it has a certain degree of anti-oxidative damage.