MANF对内质网应激诱导小肠上皮细胞损伤的保护作用  

The protective effect of MANF on intestinal epithelial cell injury induced by endoplasmic reticulum stress

作　　者：陈滢[1] 周南 王杰 倪倩倩 赵倩[4] CHEN Ying;ZHOU Nan;WANG Jie;NI Qian-qian;ZHAO Qian(School of Nursing;School of Basic Medical Sciences, Anhui Medical University, Hefei 230032, China;Dept of Obstetrics, The First Affiliated Hospital of USTC, Hefei 230032, China;Dept of Neonatology, The First Affiliated Hospital of Anhui Medical University, Hefei 230032, China)

机构地区：[1]安徽医科大学护理学院 [2]安徽医科大学基础医学院 [3]中国科学技术大学附属第一医院产科 [4]安徽医科大学第一附属医院新生儿科,安徽合肥230032

出　　处：《中国药理学通报》2022年第7期1066-1071,共6页Chinese Pharmacological Bulletin

基　　金：安徽省自然科学基金青年基金项目(No 1808085QH239);安徽医科大学护理学院科研基金青年培育项目(No hlqn2020009,hlpy20210016)。

摘　　要：目的在正常人小肠上皮细胞系FHs 74 Int中观察中脑星形胶质细胞源性神经营养因子(mesencephalic astrocyte-derived neurotrophic factor,MANF)对内质网(endoplasmic reticulum,ER)应激诱导的肠道上皮细胞损伤的保护作用。方法分别用ER应激诱导剂tunicmycin(TM)或TNF-α刺激FHs 74 Int细胞,观察内源性MANF的表达;将MANF-GFP质粒转染FHs 74 Int细胞,用免疫印迹法(Western blot)观察MANF对ER应激指标的影响;CCK8细胞增殖实验检测MANF对肠上皮细胞发生ER应激后增殖能力的影响;Western blot和流式细胞术(Flow cytometry)检测MANF对ER应激后细胞凋亡的影响。结果ER应激可以诱导小肠上皮细结论MANF通过抑制小肠上皮细胞内的ER应激,促进细胞增殖,减少细胞凋亡,从而发挥肠黏膜屏障保护作用。Aim To observe the protective effect of mesencephalic astrocyte-derived neurotrophic factor(MANF)on intestinal epithelial cells under endoplasmic reticulum stress(ER stress).Methods Normal human intestinal epithelial cell line FHs74Int was stimulated with ER stress inducers,tunicmycin(TM)and TNF-α,then the expression of endogenous MANF was observed.The recombinant plasmids MANF-GFP and GFP were transfected into FHs74Int cells individually,the transfection efficiency was observed by fluorescence microscopy,and the effect of MANF on ER stress was observed by Western blot.The effect of MANF on the proliferation of intestinal epithelial cells stimulated by TM was detected via CCK-8 assay.The effect of MANF on apoptosis after ER stress was detected by Western blot and flow cytometry.Results ER stress could induce the expression of endogenous MANF in intestinal epithelial cells.Overexpression of MANF significantly inhibited the expression of the ER stress-related proteins,Bip and CHOP,and promoted the proliferation of intestinal epithelial cells.At the same time,it could reduce the production of the proapoptotic proteins cleaved-caspase-3 and Bax,increase the expression of the antiapoptotic protein Bcl-2,and inhibit the proportion of early and late apoptosis of intestinal epithelial cells.Conclusions MANF plays a protective role in inhibiting ER stress in intestinal epithelial cells by promoting cell proliferation and reducing apoptosis.

关 键 词：MANF TM 内质网应激 小肠上皮细胞 增殖 凋亡 

分 类 号：R322.45[医药卫生—人体解剖和组织胚胎学] R329.24[医药卫生—基础医学] R329.25R329.28R574R977.6