下调PTEN对喉返神经挤压损伤大鼠的干预作用及机制研究  

Intervention effects of down-regulation of PTEN expression on rat models with RLN crush injury and related action mechanism

作　　者：王殿一[1] 武峙璇[2] WANG Dianyi;WU Shixuan(Department of ENT,The First Affiliated Hospital of Heilongjiang TCM University,Heilongjiang,Harbin 150040,China)

机构地区：[1]黑龙江中医药大学附属第一医院耳鼻喉科,哈尔滨市150040 [2]黑龙江省医院老年神经内科

出　　处：《河北医药》2021年第13期1935-1939,共5页Hebei Medical Journal

摘　　要：目的研究下调人第10号染色体缺失的磷酸酶及张力蛋白同源基因(phosphatase andtensin homolog deleted on chromosome ten,PTEN)表达对喉返神经挤压损伤模型大鼠的干预作用及相关机制。方法选取40只健康清洁级雄性大鼠,抽取30只建立喉返神经挤压损伤模型,成功27只,分为模型组、下调组、上调组,每组9只;另10只作为假手术组。建立下调PTEN慢病毒表达载体,定量PCR检测PTEN mRNA基因表达,声学改变、甲杓肌神经诱发电位图、喉返神经纤维、杓状软骨运动角度检查,病理组织观察,Western blot检测层粘连蛋白激活憐酸肌醇3激酶(phosphoinositide 3-kinase,PI3K)、神经干睫状神经营养因子(ciliary neurotrophic factor,CNTF)、Myogenin、Akt、GSK-3β相关蛋白。结果模型组、下调组、上调组PTEN mRNA表达升高,杓状软骨运动最大角度、PI3K、Akt、GSK-3β、CNTF、Myogenin相关蛋白表达降低,差异均有统计学意义(P<0.05)。下调组PTEN mRNA表达降低,杓状软骨运动最大角度、PI3K、Akt、GSK-3β、CNTF、Myogenin蛋白表达升高;上调组PTEN mRNA表达升高,杓状软骨运动最大角度、PI3K、Akt、GSK-3β、CNTF、Myogenin蛋白表达降低,差异均有统计学意义(P<0.05)。结论下调PTEN表达对喉返神经挤压损伤模型大鼠干预效果显著,下调PTEN下游PI3K、Akt、GSK-3β、CNTF、Myogenin蛋白,激活PI3K/Akt/GSK-3β通路,参与大鼠喉返神经修复过程。Objective To investigate the intervention effects of down-regulation of phosphatase andtensin homolog deleted on chromosome ten(PTEN)expression on rat models with RLN crush injury and related action mechanism.Methods Thirty out of forty healthy and clean male rats were selected to establish the RLN crush injury model.And the models were successfully established in 27 rats,who were divided into model group(n=9),down-regulation group(n=9)and up-regulation group(n=9).At the same time the other 10 rats were enrolled as sham operation group.Lentivirus expression vector of down-regulation PTEN was constructed.Moreover,quantitative PCR was used to detect the expression of PTEN mRNA.Furthermore,acoustic changes,nerve evoked potential map of dipper muscle,recurrent laryngeal nerve fibers,movement angle of arytenoid cartilage and pathological tissues were observed.In addition,Western Blot was used to detect phosphoinositide 3kinase,PI3K,ciliary neurotrophic factor,CNTF,Myogenin,Akt and GSK3β.Results The expression levels of PTEN mRNA were increased in model group,down-regulation group and up-regulation group,however,the expression levels of PI3K,Akt,GSK3β,CNTF,Myogenin-related protein and the maximum angle of dipper cartilage movement were significantly decreased(P<0.05).The expression levels of PTEN mRNA in down-regulation group were decreased,while the expression levels of PI3K,Akt,GSK3β,CNTF,Myogenin-related protein and the maximum angle of dipper cartilage movement increased.The expression of PTEN mRNA in up-regulation group increased while the expression of PI3K,Akt,GSK3β,CNTF,Myogenin-related protein and the maximum angle of dipper cartilage movement were significantly decreased(P<0.05).Conclusion The down-regulation of PTEN expression has a significant effect on the rat model of RLN crush injury.The down-regulation of PI3K,Akt,GSK3β,CNTF,myogenin protein can activate PI3K/Akt/GSK3βpathway and repair RLN.

关 键 词：人第10号染色体缺失的磷酸酶及张力蛋白同源基因 喉返神经 神经干睫状神经营养因子 

分 类 号：R246.81[医药卫生—针灸推拿学]