益生菌对力竭运动大鼠肠上皮细胞自噬的影响
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摘要
目的研究补充益生菌对力竭运动大鼠肠上皮细胞自噬水平的影响。方法将SD大鼠随机分为安静对照组、力竭运动组,力竭运动+益生菌低、中、高剂量组。各组灌胃不同剂量的益生菌溶液,6周力竭运动训练后,经腹腔注射麻醉后迅速剖腹大鼠,取远端回肠30cm。通过透射电镜观察大鼠肠上皮细胞自噬变化和测定大鼠肠上皮细胞Atg16L1、Beclin-1和LC3 mRNA及蛋白表达。结果力竭运动组大鼠肠黏膜上皮细胞可见大量的自噬体,力竭运动+益生菌低剂量组大鼠自噬体数目明显较少,而力竭运动+益生菌中剂量和力竭运动+益生菌高剂量组大鼠没有自噬体的产生。ME组大鼠肠上皮细胞Atg16L1、Beclin-1、LC3 mRNA和蛋白表达均高于安静对照组(P<0.01或P<0.05);MEYZ组和MEYG大鼠肠上皮细胞Atg16L1、Beclin-1、LC3 mRNA表达均低于ME组(P<0.01)。MEYZ组大鼠肠上皮细胞Beclin-1蛋白表达低于ME组(P<0.05)。MEYD、MEYZ和MEYG组大鼠LC3蛋白表达均低于ME组(P<0.01)。结论长时间的力竭运动诱发大鼠肠上皮细胞自噬体的产生,起到防御氧化应激致使肠上皮细胞受到损伤。通过补充益生菌后降低了Atg16L1、Beclin-1和LC3的表达。表明益生菌起到保护肠上皮细胞受到损伤和有效抑制反复力竭运动大鼠肠上皮细胞自噬的产生。
Objective To study the effect of autophagy in intestinal epithelial cells of probiotics-supplemented rats after exhaustive exercise.Methods The SD male rats were randomized into five groups on average,the quiet control(NC),the exhaustive exercise(ME),the exhaustive exercise with low/medium/high dose of probiotics(MEYD/MEYZ/MEYG).Rats in each group were intragastrically administered with normal saline or different doses of the probiotic solution,respectively.After 6 weeks-exhaustive exercise training,the rats were intraperitoneal injected of anesthesia,and rapidly followed laparotomy to take the distal ileum 30 cm.The autophagy was observed by transmission electron microscopy.The mRNA and protein expressions of Atg16 L1,Beclin-1 and LC3 were measured according to the corresponding kit method.Results A large number of autophagy were observed in the intestinal mucosal epithelial cells in ME,while it was significantly less in MEYD,and there were no autophagosomes in MEYZ and MEYG.The mRNA and protein expressions of Atg16 L1,Beclin-1 and LC3 in ME were higher than in NC(P<0.01 or P<0.05),while mRNA in MEYZ and MEYG were both lower than in ME(P<0.01).The expression of Beclin-1 protein in MEYZ was lower than that in ME(P<0.05).And the expression of LC3 protein in MEYD,MEYZ and MEYG rats were all lower than in ME(P<0.01).Conclusion Long-term exhaustive exercise induces autophagosome production in rat intestinal epithelial cells,which protects intestinal epithelial cells from oxidative stress.The expression of Atg16 L1,Beclin-1 and LC3 was reduced by supplementation with probiotics.It indicates that probiotics protect the intestinal epithelial cells from damage and effectively inhibit the production of autophagosomes from intestinal epithelial cells in rats after repeated exhaustion.
Objective To study the effect of autophagy in intestinal epithelial cells of probiotics-supplemented rats after exhaustive exercise.Methods The SD male rats were randomized into five groups on average,the quiet control(NC),the exhaustive exercise(ME),the exhaustive exercise with low/medium/high dose of probiotics(MEYD/MEYZ/MEYG).Rats in each group were intragastrically administered with normal saline or different doses of the probiotic solution,respectively.After 6 weeks-exhaustive exercise training,the rats were intraperitoneal injected of anesthesia,and rapidly followed laparotomy to take the distal ileum 30 cm.The autophagy was observed by transmission electron microscopy.The mRNA and protein expressions of Atg16 L1,Beclin-1 and LC3 were measured according to the corresponding kit method.Results A large number of autophagy were observed in the intestinal mucosal epithelial cells in ME,while it was significantly less in MEYD,and there were no autophagosomes in MEYZ and MEYG.The mRNA and protein expressions of Atg16 L1,Beclin-1 and LC3 in ME were higher than in NC(P<0.01 or P<0.05),while mRNA in MEYZ and MEYG were both lower than in ME(P<0.01).The expression of Beclin-1 protein in MEYZ was lower than that in ME(P<0.05).And the expression of LC3 protein in MEYD,MEYZ and MEYG rats were all lower than in ME(P<0.01).Conclusion Long-term exhaustive exercise induces autophagosome production in rat intestinal epithelial cells,which protects intestinal epithelial cells from oxidative stress.The expression of Atg16 L1,Beclin-1 and LC3 was reduced by supplementation with probiotics.It indicates that probiotics protect the intestinal epithelial cells from damage and effectively inhibit the production of autophagosomes from intestinal epithelial cells in rats after repeated exhaustion.
引文
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[4] Schwartz M,Regueiro M.Prevention and treatment of postoperative Crohn′s disease recurrence:an update for a new decade[J].Current Gastroenterology Reports,2011,13(1):95-100.
[5] Barbara G,Zecchi L,Barbaro R,et al.Mucosal permeability and immune activation as potential therapeutic targets of probiotics in irritable bowel syndrome[J].Journal of Clinical Gastroenterology,2012,46(Suppl):S52-S55.
[6] Chen YM,Wei L,Chiu YS,et al.Lactobacillus plantarum TWK10 supplementation improves exercise performance and increases muscle mass in mice[J].Nutrients,2016,8(4):205.
[7] Bedford TG,Tipton CM,Wilson NC,et al.Maximum Oxygen consumption of rats and its changes with various experimental procedures[J].Journal of Applied Physiology,1979,47(6):1278-1283.
[8] 莫伟彬,邱晓玲,李国峰.罗汉果甜苷对训练大鼠骨骼肌PGC-lαmRNA及蛋白表达的影响[J].现代预防医学,2017,44(1):131-134.
[9] Desoignie R,Sellin J.Propionate-initialed changes in intra-cellular pH in rabbit coloncytes[J].Gastroenterology,1994,107(2):347-356.
[10] Cadwell K,Liu JY,Brown SL,et al.A key role for autophagy and the autophagy gene Atg16l1 in mouse and human intestinal Paneth cells[J].Nature,2008,456(7219):259-263.
[11] Zhou R,Yazdi AS,Menu P,et al.A role for mitochondria in NLRP3 inflammasome activation[J].Nature,2011,469(7329):221-225.
[12] 樊冬梅,刘佳,李燕舞.益气解毒方干预实验性结肠炎大鼠肠上皮细胞自噬功能的研究[J].山东中医药大学学报,2016,40(1):65-68.
[13] 张旭东,柳娜,景明,等.湿生扁蕾(口山)酮对溃疡性结肠炎肠纤维化大鼠肠上皮细胞ATG16L1、LC3表达的影响[J].中药材,2018,41(3):720-724.
[14] Aita VM,Liang XH,Murty VV,et al.Cloning and genomic organization of beclin 1,a candidate tumor suppressor gene on chromosome 17q21[J].Genomics,1999,59(1):59-65.
[15] Kabeya Y,Mizushima N,Ueno T,et al.LC3,a mammalian homologue of yeast Apg8p,is localized in autophagosome membranes after processing[J].The EMBO Journal,2000,19(21):5720-5728.
[16] Zhang FX,Deng ZY,Li WX,et al.Activation of autophagy in rats with plateau stress-induced intestinal failure[J].International Journal of Clinical and Experimental Pathology,2015,8(2):1816-1821.
[17] 郑英,哈小琴,王怡君,等.大黄素在低氧肠应激损伤中细胞自噬的保护作用[J].陕西科技大学学报(自然科学版),2015(2):134-138.