CAJ | 학술논문

目的：观察谷氨酰胺对饥饿小鼠小肠乳脂肪球上皮生长因子E8（MFG-E8）表达的影响。方法将25只雄性C57BL/6小鼠随机分为正常对照组（NC组，n=5），全饥饿组（S组，n=5）和谷氨酰胺灌胃组（G组，n=15）。其中谷氨酰胺灌胃组根据不同的灌胃量[1g/（kg?d）、3g/（kg?d）、5g/（kg?d）]分为3个亚组G1组（n=5）、G3组（n=5）、G5组（n=5）。NC组正常饮食，饮水。S组和G组仅正常饮水。G组给予不同剂量谷氨酰胺水溶液灌胃，1次/d，0.5ml/次，共3次。NC组和S组给予生理盐水灌胃0.5ml/d。72h后脱颈法处死小鼠，取小肠组织。HE染色观察小肠绒毛发育；TUNEL法检测细胞凋亡；实时荧光定量PCR和蛋白质免疫印迹法检测小肠组织MFG-E8mRNA及MFG-E8蛋白表达。结果饥饿72h后小肠绒毛呈现萎缩状态，绒毛高度、隐窝深度及绒毛面积均明显降低；黏膜上皮细胞凋亡增加；MFG-E8在基因水平和蛋白水平表达均增加。谷氨酰胺灌胃后小肠绒毛萎缩状态改善，黏膜上皮细胞凋亡减少，MFG-E8mRNA和蛋白均较单纯饥饿组降低，且mRNA降低与谷氨酰胺灌胃量呈剂量依赖。结论 MFG-E8可作为饥饿后反映小肠代偿功能的标志，MFG-E8升高可能是小肠饥饿损伤后的一种自我保护性反应。
목적：관찰곡안선알대기아소서소장유지방구상피생장인자E8（MFG-E8）표체적영향。방법장25지웅성C57BL/6소서수궤분위정상대조조（NC조，n=5），전기아조（S조，n=5）화곡안선알관위조（G조，n=15）。기중곡안선알관위조근거불동적관위량[1g/（kg?d）、3g/（kg?d）、5g/（kg?d）]분위3개아조G1조（n=5）、G3조（n=5）、G5조（n=5）。NC조정상음식，음수。S조화G조부정상음수。G조급여불동제량곡안선알수용액관위，1차/d，0.5ml/차，공3차。NC조화S조급여생리염수관위0.5ml/d。72h후탈경법처사소서，취소장조직。HE염색관찰소장융모발육；TUNEL법검측세포조망；실시형광정량PCR화단백질면역인적법검측소장조직MFG-E8mRNA급MFG-E8단백표체。결과기아72h후소장융모정현위축상태，융모고도、은와심도급융모면적균명현강저；점막상피세포조망증가；MFG-E8재기인수평화단백수평표체균증가。곡안선알관위후소장융모위축상태개선，점막상피세포조망감소，MFG-E8mRNA화단백균교단순기아조강저，차mRNA강저여곡안선알관위량정제량의뢰。결론 MFG-E8가작위기아후반영소장대상공능적표지，MFG-E8승고가능시소장기아손상후적일충자아보호성반응。
Objective To observe the effect of glutamine on intestinal milk fat globule–EGF factor 8 expression on the hungry mice. MethodsTwenty-five C57BL/6 male mice were randomly divided into five groups: normal control group(NC,n=5),starvation group(S, n=5)and glutamine group(G,n=15). G group received Glu 1.0 or 3.0 or 5.0 g/kg by gastric feeding once a day from the day of starvation and named G1 or G3 or G5 group respectively. NC group ate food and drank water freely.S and G group drank water only and were of no food supply.The mice were sacrificed in 72 hours after starvation.The intestine specimens were sampled for histological examination by HE staining and cell apoptosis detection by TUNEL assay.The expression of MFG-E8mRNA and protein was measured by Real-Time PCR and Western Blot.ResultsStarvation resulted in damage to the intestinal mucosa. Villus heights,crypt depths and villus areas were significantly reduced and the intestine epithelial apoptosis was increased.The expression of MFG-E8 protein and mRNA were increased.Glutamine ameliorated villous atrophy and epithelial apoptosis.At the same time,glutamine decreased the MFG-E8 expression compared to S group.Conclusion As a kind of protective protein which is closely related to intestinal mucosa damage repair,mice intestinal MFG-E8 protein and mRNA expressions increase after starvation and decrease after Glutamine gastric feeding.This suggests MFG-E8 may be a type of protective response to intestinal damages.