CNTF干扰对BMSCs动脉移植缺血再灌注损伤脊髓下游信号通路STAT3/Caspase-9的影响  

Effects of CNTF Antibody Block to the Bone Marrow Mesenchymal Stem Cells Transplantated by Abdominal Aortic on the Downstream Signaling Pathways STAT3/Caspase-9 in Spinal Cord Ischemic Reperfusion Injuried Rats

作　　者：陈珊珊 易敏春 周国忠 普跃昌 胡毅 韩米华 金华[2] CHEN Shan-shan;YI Min-chun;ZHOU Guo-zhong;PU Yue-chang;HU Yi;HAN Mi-hua;JIN Hua(Dept.of Surgery,Chuxiong Medical College,Chuxiong 675000;Dept.of Anesthesiology,The Firstpeople＇s Hospital of Yunan Province,Kunming Yunnan 650031,China)

机构地区：[1]楚雄医药高等专科学校外科教研室,云南楚雄675000 [2]云南省第一人民医院麻醉科,云南昆明650031

出　　处：《昆明医科大学学报》2018年第8期5-12,共8页Journal of Kunming Medical University

基　　金：国家自然科学基金资助项目(31360246;81300973);云南省教育厅科学研究基金资助项目(2016ZZX279);云南省学术技术后备人才基金资助项目(2017HB042);云南省学术带头人基金资助项目(D-201631)

摘　　要：目的探讨CNTF干扰对肾下腹主动脉移植骨髓间充质干细胞治疗SCIRI大鼠脊髓下游信号通路STAT3/Caspase-9及损伤脊髓功能恢复的影响.方法成年SD雌性大鼠随机分组.BBB评分、CSEP和MEP法检测大鼠行为和神经电生理;IHC检测缺血脊髓腰段CNTF的表达;Western Blot、RT-PCR分析缺血脊髓腰段STAT3、p-STAT3、CNTF、Caspase-9的表达.结果对照组、移植组和Anti-CNTF干扰组大鼠BBB评分在术后各时间点均显著低于假手术组(P<0.01),MEP、CSEP潜伏期亦延长(P<0.01)、波幅亦减小(P<0.01),对照组变化最明显,Anti-CNTF干扰组次之,移植组最弱,组间均差异有统计学意义(P<0.05).术后7d,脊髓CNTF免疫阳性产物在Anti-CNTF干扰组减少(P<0.05),在对照组和移植组均增加(P<0.05),且移植组高于对照组(P<0.05).术后7d,缺血节段脊髓内CNTF、STAT3和p-STAT3 m RNA和蛋白表达在Anti-CNTF干扰组减少(P<0.05),在对照组和移植组均增加(P<0.05),且移植组高于对照组(P<0.05);而Caspase-9基因和蛋白表达仅在移植组减少(P<0.05),在Anti-CNTF干扰组和对照组均增加(P<0.01),且Anti-CNTF干扰组增加更显著(P<0.05).术后14d,Anti-CNTF干扰组脊髓内CNTF、STAT3和p-STAT3 m RNA和蛋白表达明显高于假手术组和对照组(P<0.05),而Caspase-9的表达高于假手术组却低于对照组(P<0.05),各因子的表达较移植组差异无统计学意义(P>0.05).结论 BMSCs高表达CNTF介导下游STAT3/Caspase-9信号通路促进SCIRI大鼠后肢功能恢复,而CNTF干扰可以抑制STAT3/Caspase-9信号通路,不利于BMSCs移植对SCIRI大鼠后肢功能改善.Objective To investigate the functional recovery and the effect of CNTF antibody block to thebone marrow mesenchymal stem cells （BMSCs ） transplantated by Abdominal aortic on the downstream signalingpathways STAT3/Caspase-9 in spinal cord ischemic reperfusion injury in rats.Methods Adult female SD ratswere assigned randomly to 4 groups. The neurological functional status of the animals was assessed with BBB scores,the Motor evoked potentials （MEP ） and Cortical somatosensory evoked potentials （CSEP ） .The IHC were usedto detect the the expressional changes of CNTF, then Western blot and RT-PCR were used to detect the expressionalchanges of STAT3、 p-STAT3、 CNTF and Caspase-9 in the ischemic segments of spinal cord.ResultsCompared with the sham group,in the SCIRI rats,the BBB scores were markedly decreased at all time points（ P〈0.01）, the latency and the amplitude of MEP and CSEP was longer and lower at 14d post operation （P 〈0.01）,and this change was the most significant in the control group the second in the CNTF block group,and the last in thetransplantation group,Resutts between each two groups were statistically significant （ P〈0.05） . At 7d postoperation, compared with the sham group, the immunoreactive products of CNTF were decreased in the CNTFblock group （ P〈0.05）, but were increased （P 〈0.05） in the control group and the transplantation group （ P〈0.05）, and results in the transplantation group were higher than in the control group （ P〈0.05） . At 7d postoperation, compared with the sham group, the mRNA and protein level of CNTF、STAT3、p-STAT3 weredecreased obviously in CNTF block group （ P〈0.05）, the levels were increased in the control group and thetransplantation group （ P〈0.05）, and the levels in the transplantation group were higher than that in the controlgroup （ P〈0.05）;but the mRNA and protein level of Caspase-9 were only decreased in the transplantation group（ P〈0.05）, the level was increased in the CNTF block grou

关 键 词：脊髓 腹主动脉 缺血再灌注 BMSC CNTF STAT3 

分 类 号：R681.54[医药卫生—骨科学]