低压低氧应激促进人脐带血间充质干细胞自噬  

Hypobaric Hypoxia Induces Autophagy of Umbilical Cord Mesenchymal Stem Cells

作　　者：尹姣姣 李兵 赵强[1] 张姝婷 潘耀柱[1] 白海[1] YIN Jiao-Jiao;LI Bing;ZHAO Qiang;ZHANG Shu-Ting;PAN Yao-Zhu;BAI Hai(Department of Hematology,Center of Hematologic Diseases of Chinese PLA,Lanzhou Military Command General Hospital,Lanzhou 730050,Gansu Province,Chin)

机构地区：[1]兰州军区总医院血液科全军血液病中心,甘肃兰州730050

出　　处：《中国实验血液学杂志》2018年第4期1194-1197,共4页Journal of Experimental Hematology

基　　金：国家自然科学基金资助项目(81372132;81502583)

摘　　要：目的:探讨低压低氧应激条件下人脐带血间充质干细胞(MSC)自噬活性的变化及其对细胞存活的影响。方法:采用低压低氧装置培养MSC,将压力控制在41.1 k Pa,氧气浓度控制在1%,在0、4、8、16、24和48 h收取细胞,分别采用Western blot和RT-PCR的方法检测自噬标记性蛋白LC3B的蛋白表达水平和mRNA转录水平的变化;采用自噬抑制剂羟氯喹(8μg/ml)和三甲基腺嘌呤(5 mmol/L)处理细胞,观察低压低氧条件下,抑制自噬后M SC的存活状况。结果:低压低氧培养M SC后LC3Ⅱ的蛋白表达量和mRNA转录水平在8 h时间点上调最明显;低压低氧培养MSC 8 h,自噬抑制组的细胞与对照组相比,细胞增殖率明显下降(P<0.05),Caspase-3蛋白表达量和细胞凋亡率增高。结论:低压低氧应激可以促进MSC自噬活化,该活性对MSC的存活具有保护作用。Objective： To investigate the autophagy activity changes of umbilical cord mesenchymal cells（ MSC）under hypobaric hypoxia and the effect of hypobaric hypoxia on cell viability. Methods： Umbilical cord mesenchymal cells were cultured in the chamber of hypobaric hypoxia with an air pressure of 41. 1 kPa and an oxygen density of 1%.At 0,4,8,16,24 and 48 hours,the cells were harvested for Western blot and real-time PCR to observe the expression level of the autophagy marker protein LC3B. And the cell viability under hypobaric hypoxia was evaluated after treatment with autophagy inhibitors HCQ（ 8 μg/ml） and 3 MA（ 5 mmol/L）. Results： LC3B expression in MSC at protein and mRNA levels were up-regulated significantly after being cultured under hypobaric hypoxia condition for 8 hours. And compared with the control group,inhibition of autophagy reduced cell viability while increased Caspase-3 expression and the incidence of apoptosis. Conclusion： Hypobaric hypoxia activates autophagy in MSC,and the activation of autophagy might play a protective role for cell survival.

关 键 词：低压低氧 脐带血间充质干细胞 细胞自噬 细胞凋亡 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]