动物营养学报2017，Vol.29Issue(8)：2734-2742,9.DOI:10.3969/j.issn.1006-267x.2017.08.016
谷氨酰胺对过氧化氢诱导氧化应激人结肠癌HT-29细胞损伤和凋亡的影响
Effects of Glutamine on Oxidative Damage and Apoptosis of Human Enterocyte-Like HT-29 Cells Induced by Hydrogen Peroxide
摘要
Abstract
This experiment was conducted to study effects of glutamine ( Gln) on oxidative damage and apop-tosis of human enterocyte-like HT-29 cells induced by hydrogen peroxide (H2O2), and to explore the antioxi-dant function of Gln and its mechanisms. HT-29 cells I were induced by different concentrations [ 0 ( control group), 0.5, 2.0 and 10.0 mmol/L] Gln and 0.35 mmol/L H2O2 for 12, 24 and 32 h, the superoxide dis-mutase ( SOD) activity and malondialdehyde ( MDA) content in cells were measured, and the mRNA relative expression of related to cell apoptosis gene were also assessed by RT-qPCR. Furthermore, the HT-29 cells were dyed with the Annexin V-FITC/PI double staining method, and the apoptosis of cells was determined by the flow cytometer. The results showed as follows:1) after treated 24 h, the SOD activity in 0.5 and 2.0 mmol/L Gln groups was significantly higher than that in control group ( P<0.05); after treated 32 h, the SOD activity in 0.5 and 2.0 mmol/L Gln groups was significantly higher than that in control group (P<0.05), and the MDA content was significantly lower than that in control group ( P<0.05) . 2) After treated 12 h, there were no significant differences on the mRNA relative expressions of cysteinyl aspartate specific proteinase-3 (Caspase-3) and B cell lymphoma/leukemia-2 associated X protein (Bax) among all groups (P>0.05). Compared with the control group, the mRNA relative expression of nuclear factor kappa B ( NF-κB ) in 0.5 mmol/L Gln group was significantly decreased (P<0.05), the mRNA relative expression of B cell lym-phoma/leukemia-2 (Bcl-2) in 0.5 and 2.0 mmol/L Gln groups was significantly increased (P<0.05). After treated 24 h, compared with the control group, the mRNA relative expressions of Caspase-3, NF-κB and Bax in 0.5, 2.0 and 10.0 mmol/L Gln groups were significantly decreased (P<0.05), while the mRNA relative expression of Bcl-2 was significantly increased ( P<0. 05 ) . After treated 32 h, compared with the control group, the mRNA relative expressions of FAS, Caspase-3, NF-κB and Bax in 2.0 mmol/L Gln group were significantly decreased (P<0.05), while the mRNA relative expression of Bcl-2 was significantly increased (P<0.05); the mRNA relative expressions of FAS, Caspase-3, NF-κB and Bax in 10.0 mmol/L Gln group were significantly decreased (P<0.05). 3) After treated 24 h, compared with the control group, the living cells number in Gln groups increased by 5.32% to 11.97%, and the necrotic cells number decreased by 6.75%to 12.66%. After treated 32 h, compared with the control group, the living cells number in Gln groups in-creased by 1.39% to 7.63%, and the necrotic cells number decreased by 3.40% to 4.57%. It is concluded that glutamine can restrain the oxidant damage of HT-29 induced by H2O2 and decrease the cells apoptosis.关键词
谷氨酰胺/氧化损伤/细胞凋亡/HT-29细胞/过氧化氢Key words
glutamine/oxidative damage/cell apoptosis/HT-29 cell/hydrogen peroxide分类
农业科技引用本文复制引用
肖英平,桂国弘,陈安国,何祥祥,李开锋,杨华..谷氨酰胺对过氧化氢诱导氧化应激人结肠癌HT-29细胞损伤和凋亡的影响[J].动物营养学报,2017,29(8):2734-2742,9.基金项目
国家自然科学基金(31402083) （31402083）
浙江省自然科学基金重点项目(LZ15C170001) （LZ15C170001）
