脊髓钝挫伤后慢病毒干扰水通道蛋白4对大鼠运动功能及Rho表达的影响  

Effect of Lentivirus-Mediated Inhibition of Aquaporin 4 on Motor Function and Rho Expression in Rats with Spinal Cord Contusion

作　　者：何正云 夏登桃 肖颜 唐玉玲 卫家豪 徐轩 李沁轩 张晓[1] 荣成[1] He Zhengyun;Xia Dengtao;Xiao Yan;Tang Yuling;Wei Jiahao;Xu Xuan;Li Qinxuan;Zhang Xiao;Rong Cheng(Experimental Teaching Center,School of Basic Medical Sciences,Chengdu Medical College,Chengdu 610500,China)

机构地区：[1]成都医学院基础医学院实验教学中心,成都610500

出　　处：《成都医学院学报》2023年第1期39-44,共6页Journal of Chengdu Medical College

基　　金：四川省大学生创新创业训练项目(No:201813705063)。

摘　　要：目的 探讨脊髓钝挫伤(SCC)后,慢病毒干扰水通道蛋白4(AQP4)对大鼠运动功能的影响,并观察凋亡相关因子Rho表达的变化。方法 将SD大鼠随机分为假手术组(Sham组)、生理盐水组(SCC组)、空载组(Vector组)和慢病毒组(AQP4RNAi组,制备慢病毒AQP4干扰模型),每组32只。SCC组、AQP4RNAi组及Vector组均采用改良Allen’s法制备SCC模型,术后12 h及术后1、3 d,采用大鼠脊髓损伤(BBB)评分评估4组后肢运动功能;采用实时荧光定量聚合酶链反应(RT-PCR)检测4组脊髓组织中rho的表达;采用免疫组织化学方法检测Rho在4组脊髓组织中的表达;生物信息学分析预测AQP4与Rho之间的相关性。结果 运动功能评分结果显示,术后12 h SCC组、Vector组和AQP4 RNAi组BBB评分均为0分,术后3 d AQP4 RNAi组BBB评分较Vector组高(P<0.05)。GeneMANIA结果显示,AQP4与Rho存在间接共表达关系。RT-PCR和免疫组织化学结果显示,术后12 h及术后1、3 d AQP4RNAi组脊髓组织中rho表达均低于Vector组(P<0.05);SCC组脊髓组织中rho表达均高于Sham组(P<0.05)。结论 大鼠SCC后脊髓组织中Rho表达上调,而干扰AQP4能使其Rho表达下调,减少神经细胞凋亡,有利于大鼠运动功能恢复。Objective To investigate the effect of lentivirus-mediated inhibition of aquaporin 4(AQP4) on motor function of rats with spinal cord contusion(SCC),and observe the expression change of Rho,an apoptosis related factor.Methods Sprague Dawley(SD) rats were randomly divided into sham operation group(Sham group),normal saline group(SCC group),empty load group(Vector group),and lentivirus interfering group(AQP4 RNAi group),with 32 rats in each group. SCC rat models were prepared by modified Allen’s test. At 12 h,1 d,and 3 d after modeling,the hindlimb motor function of rats was evaluated by Basso,Beattie and Bresnahan locomotor rating scale(BBB),the expression of rho in the spinal cord tissues of the four groups was detected by real-time fluorescence quantitative polymerase chain reaction(RT-PCR), the expression of Rho in the spinal cord tissues of the four groups was detected by immunohistochemistry. And the correlation between AQP4 and Rho was predicted by GeneMAINA biological information database. Results SCC group,Vector group and AQP4 RNAi group scored 0 in BBB at 12 h after operation,and the BBB score of AQP4 RNAi group at 3 d after operation was higher than that of Vector group(P< 0.05). GeneMANIA analysis results showed that there was an indirect co-expression relationship between AQP4 and Rho. RT-PCR and immunohistochemistry results showed that at 12 h,1 d and 3 d after operation,the expression of rho in the spinal cord tissue of AQP4 RNAi group was lower than that of Vector group(P< 0.05),and the expression of rho in the spinal cord tissue of SCC group was higher than that of Sham group(P< 0.05). Conclusion The expression of apoptosis-related factor Rho is up-regulated in the spinal cord tissue of SCC rats.Interference with AQP4 can down-regulate the expression of Rho and reduce the apoptosis of nerve cells,which is beneficial to the recovery of motor function in SCC rats.

关 键 词：脊髓钝挫伤 慢病毒干扰 水通道蛋白4 凋亡 

分 类 号：R34[医药卫生—基础医学]