胎盘间充质干细胞低氧培养液对肠黏膜上皮细胞氧化应激损伤的保护作用  

Protection Effect of Placenta Mesenchymal Stem Cells Hypoxia Culture Medium on Oxidative Stress-induced Injury of Intestinal Mucosal Epithelial Cells

作　　者：杜莉莉[1] 吕润潇[2] 杨晓漪[3] 辛娜[1] 马廷贤[1] 

机构地区：[1]中国医科大学基础医学院病理生理学教研室,沈阳110013 [2]第二军医大学附属长海医院研究生队、第二军医大学研究生队,上海200433 [3]中国医科大学95期临床医学7年制,沈阳110013

出　　处：《中国医科大学学报》2016年第2期131-135,140,共6页Journal of China Medical University

基　　金：辽宁省科学技术厅2015年博士科研启动基金计划(201501003)

摘　　要：目的研究胎盘间充质干细胞(p MSCs)低氧培养液对肠黏膜上皮细胞氧化应激损伤的作用。方法组织块培养法提取p MSCs,流式细胞术检测表面标志,ELISA检测p MSCs常氧(21%O_2)或低氧(1%O_2)培养5 d后培养液中胰岛素样生长因子1(IGF-1)的含量。人结肠腺癌细胞(Caco2)经100μmol/L H_2O_2处理12 h后分别培养于正常培养液、p MSCs常氧培养液或p MSCs低氧培养液,5 d后台盼蓝染色测细胞存活情况,MTT测细胞增殖,Annexin V-FITC/PI双染测细胞凋亡。结果 p MSCs阳性表达CD29,CD44。p MSCs低氧培养液中的IGF-1明显多于p MSCs常氧培养液(P<0.05)。与p MSCs常氧培养液相比,p MSCs低氧培养液中H_2O_2处理的Caco2存活细胞多,增殖快,凋亡少(P<0.05)。向p MSCs低氧培养液中加入IGF-1抗体后,细胞增殖减慢,凋亡增多(P<0.05)。结论 p MSCs低氧培养液对H_2O_2处理的Caco2具有保护作用。机制与IGF-1促进细胞增殖、抑制凋亡相关。Objective To study the effect of placenta mesenchymal stem cells （pMSCs） hypoxia culture medium on the oxidative stress-induced injury of intestinal mucosal epithelial cells. Methods pMSCs were extracted by tissue culture method and identified by flow cytometry. The IGF- 1 content in the culture medium of pMSCs cultured under both normal oxygen （21% 02） and hypoxia （ 1% 02） were detected by ELISA. Caco2 were treated with H2O2 （ 100 μmol/L ）for 12 h and then cultured in normal culture medium （NM）, pMSCs normoxia culture medium （pMSCs-NCM ） or pMSCs hypoxia culture medium （pMSCs-HCM） for 5 d respectively. Cell survival was detected by trypan blue staining, cell proliferation was detected with MTT and cell apoptosis was detected with Annexin V-FITC/PI. Results pMSCs positively expressed CD29 and CD44. IGF- 1 in pMSCs- HCM was significantly more than that in pMSCs-NCM （P 〈 0.05）. Compared with pMSCs-NCM, H2O2-treated Caco2 in pMSCs-HCM contained more living cells. Their proliferation was faster and apoptosis was slower （P 〈 0.05 ）. After the addition of IGF- 1 antibody to pMSCs-HCM, the prolif- eration of H2O2-treated Caco2 decreased and the apoptosis increased （P 〈 0.05 ）. Conclusion pMSCs-HCM had a protective effect on H2O2-treated Caco2, and the mechanism may be related to the IGF- 1 mediated cell proliferation and cell apoptosis inhibition.

关 键 词：胎盘间充质干细胞 低氧 氧化应激 人结肠腺癌细胞 

分 类 号：R574.5[医药卫生—消化系统]