胰高血糖素样肽-2对小鼠肠黏膜干细胞LGR5表达的影响及对肠道黏膜的保护作用  

Effect of glucagon-like peptides-2 on LGR5 expression of intestinal mucosal stem cell and its protective role on intestinal mucosa in mice

作　　者：孙莹[1] 王丽杰[1] 

机构地区：[1]中国医科大学附属盛京医院PICU,沈阳110004

出　　处：《中国小儿急救医学》2014年第12期777-781,共5页Chinese Pediatric Emergency Medicine

基　　金：辽宁省自然科学基金（20102258）

摘　　要：目的 研究LGR5肠道干细胞在内毒素血症肠黏膜损伤修复中的作用,是否可以通过调节干细胞的增殖分化来促进肠黏膜的修复.方法 21日龄健康Wistar幼鼠,随机分成对照组、脂多糖（lipopolysaccharide,LPS）组和胰高血糖素样肽-2（glucagon-like peptides 2,GLP-2）组.LPS组及GLP-2组腹腔注射LPS 5 mg/kg,GLP-2组腹腔注射LPS 1 h后腹腔注射GLP-2 250 μg/kg;对照组腹腔注射生理盐水1 ml/kg;在LPS注射后6h、24 h及72 h取末端回肠.光镜及电镜观察各组回肠上皮结构改变,免疫组织化学、RT-PCR方法检测肠道干细胞标记物LGR5的表达.结果 LPS注射后6h,LPS组及GLP-2组大体可见肠管充血、水肿.光镜下可见绒毛断裂、倒伏,炎性细胞浸润,腔内可见渗出液,24h及72h LPS组及GLP-2组肠黏膜损伤逐渐修复,绒毛逐渐生长,GLP-2组较LPS组修复明显,72h GLP-2组上皮基本恢复正常.免疫组织化学可见LGR5抗体标记在隐窝内,LPS组及GLP-2组在绒毛底部及绒毛处可见LGR5抗体标记,GLP-2组表达多于LPS组.RT-PCR检测,LPS组LGR5 mRNA表达6 h（0.13±0.05）、24h（0.16±0.05）、72 h（0.16±0.04）均明显高于对照组（0.12 ±0.03）,差异均有统计学意义（P均＜0.05）;GLP-2组LGR5 mRNA表达6h（0.52±0.09）、24 h（0.73 ±0.14）、72 h（0.48 ±0.24）,明显高于LPS组,差异均有统计学意义（P均＜0.05）.结论 内毒素血症肠上皮黏膜炎症损伤后,肠上皮干细胞增殖分化,可能参与修复炎症损伤后的肠黏膜;外源性给予GLP-2可促进内毒素血症肠黏膜损伤的修复.Objective To explore the role of LGR5 positive intestinal stem cells in repairing damage of intestinal mucosa resulting from endotoxemia and to determine whether the damaged intestinal mucosa can be repaired by regulating the proliferation and differentiation of intestinal stem cells.Methods Wistar rats were randomly divided into control group,lipopolysaccharide（LPS） group,and glucagon-like peptides 2（GLP-2） group.LPS was injected interaperitoneally into rats of the LPS group and GLP-2 group at a dose of 5 mg/kg（1 ml/kg） ; saline（1 ml/kg） was injected into the rats of control group.GLP-2 250 μg/kg（1 ml/kg） was rnjected into the rats of GLP-2 group 1 hour after the LPS injection.The terminal ilea were collected from 8 rats in each group at 6 h,24 h,and 72 h post-LPS injection.Structrral changes in the intestinal epthelium of every group were observed under the light microscope and electron microscope.The expression of LGR5 in intestinal stem cell was detected by immunohistochemical method and RT-PCR.Results In general,intestinal edema and hyperemia was observed in both LPS and GLP-2 groups at 6 h.Fracture and lodging of villi,infiltration of inflammatory cells,and visible exudate within the cavity were observed under light microscopy.The inflammatory injury was less severe in the GLP-2 group than in the LPS group at 6 h.The injury of intestina mucosa gradually repaired between 24 h and 72 h after injection of LPS in both LPS group and GLP-2 group.Importantly,samples taken from GLP-2 group and LPS group at the same time point indicated that the GLP-2 group recovered significantly better than the LPS group.And expressions of LGR5 mRNA in GLP-2 group at 6h,24h,72 h（0.13 ＆#177;0.05,0.16＆#177;0.05,0.16＆#177;0.04） were significantly higher than those in LPS group respectively （0.52 ＆#177;0.09,0.73 ＆#177;0.14,0.48 ＆#177;0.24）,as shown by RT-PCR,and than that in control group（0.12 ＆#177;0.03） （P 〈 0.05）.Conclusion Exogenous GLP-2 may facilitate intestinal stem cell prol

关 键 词：内毒素血症 肠黏膜 干细胞 大鼠 

分 类 号：R574[医药卫生—消化系统]