七氟烷预处理和miR-29b-3p对大鼠脊髓缺血/再灌注损伤的影响  

Effects of sevoflurane preconditioning and miR-29b-3p on spinal cord ischemia/reperfusion injury in rats

作　　者：马克军 方波[1] 陈凤收[1] 李哲[1] 贾慧[1] 马虹[1] MA Ke-jun;FANG Bo;CHEN Feng-shou;LI Zhe;JIA Hui;MA Hong(Dept of Anesthesiology, First Affliated Hospital, China Medical University, Shenyang 110001, China)

机构地区：[1]中国医科大学附属第一医院麻醉科,辽宁沈阳110001

出　　处：《中国药理学通报》2020年第10期1368-1373,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81771342)。

摘　　要：目的探讨七氟烷预处理和miR-29b-3p对大鼠脊髓缺血/再灌注损伤(spinal cord ischemia/reperfusion injury,SCIRI)的影响及相关机制。方法通过夹闭主动脉弓14 min建立大鼠SCIRI模型。缺血/再灌注(IR)前将大鼠暴露于2.4%(1 Mac)七氟烷中1 h,建立七氟烷预处理模型。通过鞘内注射mimic质粒过表达miR-29b-3p。采用Tarlov评分、伊文思蓝(EB)、HE、TUNEL染色评估脊髓损伤;Real-time qPCR检测miR-29b-3p和BMF mRNA表达;Western blot测定凋亡相关蛋白BCL-2蛋白修饰因子(BMF)、Bax、和cleaved caspase-3表达。结果与假手术组相比,IR组Tarlov评分降低,血脊髓屏障(BSCB)通透性增加,健存运动神经元减少,凋亡细胞增多,miR-29b-3p表达减少,BMF、Bax、和cleaved caspase-3表达增加(P<0.05)。和IR组相比,七氟烷预处理或过表达miR-29b-3p时,IR大鼠的Tarlov评分升高,BSCB通透性降低,健存运动神经元增加,细胞凋亡率减少,miR-29b-3p表达增加,BMF、Bax、和cleaved caspase-3表达降低(P<0.05)。结论miR-29b-3p参与调控SCIRI;七氟烷预处理保护大鼠SCIRI,其机制可能与miR-29b-3p相关。Aim To investigate the effects of sevoflurane preconditioning and miR-29b-3p on spinal cord ischemia/reperfusion injury(SCIRI)in rats.Methods The ischemia/reperfusion(IR)model was estab-lished by clamping the aortic arch for 14 minutes.Rats were exposed to 2.4%(1 Mac)sevoflurane for 1 h before SCIRI to establish the sevoflurane preconditioning model.miR-29b-3p was overexpressed by intrathecal injection of the mimic plasmid.Tarlov scores,Evans blue(EB),HE and TUNEL staining were used to evaluate spinal cord function and injury in rats.The expressions of miR-29b-3p and BMF mRNA were evaluated by Real-time qPCR.The protein expression of BCL-2 modifying factor(BMF),Bax and cleaved casepase 3 was detected by Western blot.Results Compared with sham operation group,IR decreased Tarlov scores,increased the permeability of blood-spinal barrier(BSCB),reduced the number of intact motor neurons,increased the apoptosis induced by IR,inhibited the expression of miR-29b-3p,and promoted the expression of BMF,Bax,and cleaved casepase 3(P<0.05).Compared with IR group,sevoflurane pretreated or over-expressed with miR-29b-3p significantly increased the Tarlov scores of IR rats,reduced the permeability of BSCB,increased the number of intact motor neurons,and decreased the apoptotic rate induced by IR(P<0.05).The expression of miR-29b-3p increased significantly,while the expression of BMF,Bax,and cleaved casepase 3 decreased(P<0.05).Conclusions miR-29b-3p can regulate SCIRI;sevoflurane preconditioning may protect SCIRI,and the mechanism may be related to miR-29b-3p.

关 键 词：七氟烷预处理 miR-29b-3p 缺血/再灌注损伤 脊髓 凋亡 BCL-2蛋白修饰因子 

分 类 号：R-332[医药卫生] R322.81