不同体积分数氧气预处理人羊膜间充质干细胞的生物学特性  

Biological characteristics of human amniotic mesenchymal stem cells pretreated with different volume fractions of oxygen

作　　者：唐建宏 张霓霓 黄桂林 龙元铸 崔田宁 骆勤亮 郎家婵 代敏 张立刚 Tang Jianhong;Zhang Nini;Huang Guilin;Long Yuanzhu;Cui Tianning;Luo Qinliang;Lang Jiachan;Dai Min;Zhang Ligang(Zunyi Medical University,Zunyi 563000,Guizhou Province,China;Department of Oral and Maxillofacial Surgery,Affiliated Stomatological Hospital of Zunyi Medical University,Zunyi 563000,Guizhou Province,China)

机构地区：[1]遵义医科大学,贵州省遵义市563000 [2]遵义医科大学附属口腔医院口腔颌面外科,贵州省遵义市563000

出　　处：《中国组织工程研究》2023年第15期2318-2324,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81860198),项目负责人:张霓霓;国家自然科学基金(81960204),项目负责人:黄桂林;遵义医科大学硕士启动基金(KY2019-1),项目负责人:代敏;贵州省卫健委基金(gzwjkj2020-1-165),项目负责人:张立刚;遵义医科大学“未来临床名医”项目(20211017),项目负责人:张霓霓。

摘　　要：背景:近年来,干细胞在组织损伤修复中发挥着重要作用,其中间充质干细胞移植应用最为广泛,间充质干细胞可以通过旁分泌细胞因子促进损伤组织的修复,低氧预处理可以改善间充质干细胞的生物学特性,因而,低氧预处理的人羊膜间充质干细胞可能在组织损伤修复中起到更好的治疗作用。目的:探讨不同体积分数氧气预处理对人羊膜间充质干细胞增殖、抗凋亡能力、旁分泌作用的影响。方法:利用酶消化法获得原代人羊膜间充质干细胞,将第3代人羊膜间充质干细胞通过不同体积分数氧气进行预处理48 h,随机分为1%,3%,5%,10%低氧预处理组和常氧对照组体积分数21%氧气,倒置显微镜下观察人羊膜间充质干细胞的形态,CCK-8检测人羊膜间充质干细胞的增殖能力,Annexin V-FITC/PI试剂盒检测人羊膜间充质干细胞的凋亡情况,实时定量PCR法检测人羊膜间充质干细胞凋亡基因和缺氧诱导因子的表达,酶联免疫吸附法检测人羊膜间充质干细胞分泌的细胞因子水平。结果与结论:显微镜下观察不同体积分数氧气预处理组人羊膜间充质干细胞的形态无明显差异,低氧预处理能提高人羊膜间充质干细胞的增殖、抗凋亡和旁分泌细胞因子的能力,且体积分数1%氧气预处理效果优于体积分数3%,5%,10%,21%氧气预处理。BACKGROUND:In recent years,stem cell plays an important role in the repair of tissue injury,among which mesenchymal stem cell transplantation is the most widely used.Mesenchymal stem cells can secrete cytokines through paracrine effect to promote the repair of damaged tissue.Hypoxia pretreatment can improve the biological characteristics of mesenchymal stem cells.Hypoxic preconditioning of human amniotic mesenchymal stem cells may play a better therapeutic role in tissue injury repair.OBJECTIVE:To study the effects of different hypoxic pretreatment on the proliferation,antiapoptotic ability,and parocrine effect of human amniotic mesenchymal stem cells.METHODS:The primary human amniotic mesenchymal stem cells were obtained by enzyme digestion method.Passage 3 human amniotic mesenchymal stem cells were pretreated with different hypoxic concentrations for 48 hours and randomly divided into hypoxic pretreatment group(1%O_(2),3%O_(2),5%O_(2),and 10%O_(2))and control group(21%O_(2)).The morphology of human amniotic mesenchymal stem cells was observed under an inverted microscope.The proliferation ability of human amniotic mesenchymal stem cells was detected by CCK-8 assay.Annexin V-FITC/PI kit was used to detect the apoptosis of human amniotic mesenchymal stem cells.The apoptosis gene and hypoxia-inducible factor of human amniotic mesenchymal stem cells were detected by real-time quantitative PCR.The cytokine secretion capacity of human amniotic mesenchymal stem cells in each group was detected by enzyme-linked immunosorbent assay.RESULTS AND CONCLUSION:Under the microscope,the morphology of human amniotic mesenchymal stem cells in different hypoxic pretreatment groups showed no significant difference.Hypoxic pretreatment could enhance the proliferation,anti-apoptotic and paracrine cytokine capacity of human amniotic mesenchymal stem cells.Pretreatment effect of 1%O_(2) concentration was better than that of 3%O_(2),5%O_(2),10%O_(2) and 21%O_(2).

关 键 词：人羊膜间充质干细胞 低氧 预处理 生物学特征 

分 类 号：R459.9[医药卫生—治疗学] R394.2[医药卫生—临床医学] R542.2