蛋白激酶D1对骨髓间充质干细胞中CXCR4的调控作用  

Regulation of protein kinase D1 on CXCR4 in bone mesenchymal stem cells

作　　者：杨雷[1,2] 刘萍 刘暖 毛秉豫 YANG Lei;LIU Ping;LIU Nuan;MAO Bingyu(Henan Key Laboratory of Zhang ZhongJing Formulae and Herbs for Immunoregulation,Nanyang Institute of Technology,Nanyang 473004,China;Zhang ZhongJing School of Chinese Medicine,Nanyang Institute of Technology,Nanyang 473004,China)

机构地区：[1]南阳理工学院河南省张仲景方药与免疫调节重点实验室,河南南阳473004 [2]南阳理工学院张仲景国医国药学院,河南南阳473004

出　　处：《暨南大学学报（自然科学与医学版）》2019年第4期279-287,共9页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：国家自然科学基金项目(81873106;81473438);河南省高等学校重点研究项目(19A360030;18B360011)

摘　　要：目的:探讨蛋白激酶D1(PKD1)对骨髓间充质干细胞(BMSCs)存活、增殖、迁移和凋亡的影响,并分析PKD1对趋化因子受体4(CXCR4)表达的调控作用.方法:从Wistar大鼠股骨和胫骨中分离并收集BMSCs细胞,体外培养鉴定.实验分为5组:对照组(Control)、氧糖剥夺-复氧组(OGD)、PKD1组、AMD3100(CXCR4的特异性阻断剂)组和AMD3100-PKD1组.CCK-8法检测BMSCs的存活率,Tunel法检测BMSCs的凋亡情况,BrdU法检测BMSCs的增殖能力,Transwell法检测BMSCs的迁移能力,RT-PCR法检测BMSCs中CXCR4的mRNA表达,Western blot法检测CXCR4的蛋白表达.结果:PKD1预处理可明显提升BMSCs的存活、增殖和迁移能力(P<0.05),减少BMSCs的凋亡数量(P<0.05);并上调BMSCs中CXCR4 mRNA和蛋白的表达(P<0.05).阻断剂AMD3100可降低PKD1预处理后BMSCs的存活、增殖和迁移能力,增加BMSCs的凋亡数量,下调BMSCs中CXCR4 mRNA和蛋白的表达,从而减弱PKD1对BMSCs的预处理作用.结论:PKD1可能通过调控CXCR4的表达发挥抗氧糖剥夺-复氧作用:减少BMSCs的凋亡,提高BMSCs的存活、增殖和迁移能力.Objective:To investigate the effects of protein kinase D1(PKD1)on the survival,proliferation,migration,and apoptosis of bone marrow mesenchymal stem cells(BMSCs),and explore its regulation on the expression of chemokine receptor 4(CXCR4).Methods:BMSCs cells were isolated from the femur and tibia of Wistar rats,then cultured and identified.The grouping of the experiment was set as follow:a control group,an oxygen-glucose deprivation-reoxygenation group(OGD),a PKD1 group,an AMD3100(CXCR4 specific blocker)group,and an AMD3100-PKD1 group.The CCK-8 method was used to determine the viability of BMSCs;the Tunel method was used to determine the apoptosis of BMSCs;the BrdU method was used to access the proliferation of BMSCs;the Transwell method was used to determine the migration of BMSCs;the reverse transcription PCR(RT-PCR)method was used to quantify the expression of CXCR4 mRNA in BMSCs;and the western blotting method was used to quantify the expression of CXCR4 protein in BMSCs.Results:PKD1 pretreatment significantly increased the viability,proliferation,and migration,decreased apoptosis,and increased mRNA and protein expression of CXCR4 in BMSCs,whereas the blocker AMD3100 reversed all the effect of PKD1 pretreatment(all P<0.05).Conclusion:PKD1 improves the viability,proliferation,and migration,and suppresses apoptosis of BMSCs might through the regulation of the expression of CXCR4,thus decreased the influence of OGD reoxygenation in BMSCs.

关 键 词：蛋白激酶D1 心肌梗死 骨髓间充质干细胞 趋化因子受体4 氧糖剥夺 

分 类 号：R322.11[医药卫生—人体解剖和组织胚胎学] R322.12[医药卫生—基础医学]