丙氨酰-谷氨酰胺预处理对氧化应激猪小肠上皮细胞活力和自噬的影响  

Effects of Pre-Adding Alanyl-Glutamine Dipeptide on Porcine Intestinal Epithelial Cells Viability and Autophagy Under Oxidative Stress

作　　者：辛向荣[1] 贺琴[1] 游金明[1] 叶亚玲[1] 邓宸玺 XIN Xiang-rong;HE Qin;YOU Jin-ming;YE Ya-ling;DENG Chen-xi(College of Animal Science and Technology,Key Laboratory of Animal Nutrition,Nutrition Feed Development Engineering Center,Jiangxi Agricultural University,Jiangxi Nanchang 330045,China)

机构地区：[1]江西农业大学动物科技学院江西省动物营养重点实验室江西省营养饲料开发工程中心,江西南昌330045

出　　处：《中国畜牧杂志》2018年第8期91-95,共5页Chinese Journal of Animal Science

基　　金：国家自然科学基金(31360554);江西省青年科学家培养计划项目(2011BCB23008)

摘　　要：试验旨在研究预添加丙氨酰-谷氨酰胺(Ala-Gln)对过氧化氢诱导下的小肠上皮细胞IPEC-1的影响。采用单因子试验设计,向IPEC-1细胞培养基中分别添加0、0.25、0.50、1.00、2.00、4.00 mmol/L Ala-Gln,预处理20 h,然后利用400μmol/L H2O2诱导细胞4 h,创建氧化应激模型。试验共设6个处理,每个处理8个重复。结果表明:IPEC-1细胞活力随Ala-Gln预处理水平的提高而提高(P<0.05);且在Ala-Gln预处理水平为2.00 mmol/L时细胞活力达最大值,4.0 mmol/L水平下反而使细胞活力下降(P<0.05),但仍高于0.50、1.00 mmol/L Ala-Gln水平下IPEC-1细胞活力;添加Ala-Gln提高了氧化应激状态IPEC-1谷胱甘肽过氧化物酶(GSH-Px)和超氧化物歧化酶(SOD)活性(P<0.05),降低了丙二醛(MDA)含量(P<0.05);细胞SOD活性呈曲线变化,在Ala-Gln预处理水平为0.25、0.50 mmol/L时活性较高,之后随Ala-Gln预处理水平提高而降低(P<0.05);在Ala-Gln预处理水平为1.00 mmol/L时,MDA含量趋于稳定;添加Ala-Gln后,随Ala-Gln水平细胞质中阳性荧光信号不断增强,且在Ala-Gln为1.00、2.00 mmol/L预处理水平下较为明显;4.00 mmol/L Ala-Gln水平下IPEC-1同对照组相比密度显著增加,且结构完整度相似,阳性荧光信号无显著差异,但同2.00 mmol/L AlaGln添加水平相比细胞密度和生物膜结构完整度提高,胞内荧光信号有减少趋势。由此可得,预添加Ala-Gln可显著提高氧化应激状态下IPEC-1细胞中GSH-Px、SOD酶活性,显著降低脂质过氧化产物MDA含量,缓解氧化应激;在IPEC-1细胞中预添加Ala-Gln能通过增强细胞自噬用以缓解细胞氧化应激。This study was conducted to evaluate the effects of pre-adding alanyl-glutamine dipeptide on porcine intestinal epithelial cells（IPEC-1） under H2 O2 oxidative stress. This experiment was designed by using single factor randomized block design, IPEC-1 cells were cultured for 24 h in Gln-free medium containing 0, 0.25, 0.5, 1.0, 2.0 or 4.0 mmol/L AlaGln, Then All the cells were induced with 400 μmol/L H2 O2 for 4 h, an oxidative stress model was established. A total of 6 treatments were performed in this study, each treatment with 8 replicates. Results showed that：（1） With the increase of Ala-Gln, the viability of IPEC-1 increased（P0.05）, which reached highest at 2.0 mmol/L Ala-Gln, whereas the viability of IPEC-1 decreased with increasing Ala-Gln at 4.0 mmol/L（P0.05）, but it still was higher than that at 0.5 or 1.0 mmol/L Ala-Gln.（2） Supplemental Ala-Gln improved the activities of glutathione peroxidase（GSH-Px） and superoxide dismutases（SOD）, and reduced malondialdehyde content（P0.05）. The activity of SOD showed a curve change, and the activity was the highest when Ala-Gln at 0.25 mmol/L or 0.5 mmol/L, and then decreased with the increase of Ala-Gln（P0.05）. While the malondialdehyde content reached stable at 1.0 mmol/L Ala-Gln.（3） With the increase of Ala-Gln, the positive fl uorescence signal in the cytoplasm was continuously enhanced, and it became more obvious at 1.0 mmol/L or 2.0 mmol/L of Ala-Gln. At 4.0 mmol/L Ala-Gln, the density of IPEC-1 increased signifi cantly compared with the control treatment, but the structural integrity was similar and there was no signifi cant difference in the positive fl uorescence signal. However, the cell density and the integrity of the biofi lm structure were increased compared with the 2.0 mmol/L Ala-Gln, and the intracellular fl uorescence signal had a decreasing trend. It can be concluded that pre-addition of Ala-Gln can signifi cantly increase the activity of GSHPx and SOD enzymes in IPEC-1 cells under oxidative stress, signifi

关 键 词：丙氨酰-谷氨酰胺 小肠上皮细胞IPEC-1 氧化应激 细胞活力 自噬 

分 类 号：S828.5[农业科学—畜牧学]