丙泊酚通过调节过氧化物氧化还原蛋白-5的表达减轻氯胺酮麻醉中神经细胞凋亡的研究  

Propofol alleviates apoptosis of nerve cells in ketamine-induced anesthesia by regulating the expression of Peroxiredoxin 5

作　　者：曹艳丽[1] 李毅[2] 宋卓悦 李广恒 庆淑梅[1] 薛金虎[1] Cao Yanli;Li Yi;Song Zhuoyue;Li Guangheng;Qing Shumei;Xue Jinhu(Department of Anesthesiology,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China;Department of Orthopaedics,the Second Affiliated Hospital of Zhengzhou University,Zhengzhou 45000,China;Department of Orthopaedics,Zhengzhou Orthopedic Hospital,Zhengzhou 450000,China;Department of Orthopaedics,Shenzhen Renmin Hospital,Shenzhen 518000,China)

机构地区：[1]郑州大学第一附属医院麻醉科,450052 [2]郑州大学第二附属医院骨科,450000 [3]郑州市骨科医院骨科,450000 [4]深圳市人民医院骨科,518000

出　　处：《中华实验外科杂志》2020年第9期1638-1642,共5页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金面上项目(81472136);河南省科技厅科技攻关项目(172102310080)。

摘　　要：目的探讨过氧化物氧化还原蛋白-5(PRDX5)在丙泊酚减轻氯胺酮致神经细胞凋亡中的机制。方法2018年3月至2019年1月,选择健康7 d龄Sprague-Dawley(SD)雄性幼鼠(北京维通利华实验动物公司)80只,将幼鼠随机分为4组(n=20),对照组腹腔注射0.9%生理盐水1 ml(对照组),实验A组腹腔注射80 mg/kg氯胺酮1 ml(氯胺酮组),实验B组腹腔注射80 mg/kg丙泊酚1 ml(丙泊酚组),实验C组腹腔注射(80 mg/kg氯胺酮+80 mg/kg丙泊酚)共1 ml(丙泊酚+氯胺酮组)。大鼠苏醒后继续饲养3周后行跳台实验(n=10),测试完毕后处死取海马组织应用免疫组织化学检测PRDX5含量(n=10)。体外培养条件下,采用孕18 d清洁级SD大鼠(北京维通利华实验动物公司,合格证编号为218764758)进行原代海马神经元培养,分4组:原代海马神经元细胞(DIV)+蒸馏水组(DIV+D组)、原代海马神经元细胞(DIV)+氯胺酮组(DIV+K组)、PRDX5基因沉默原代海马神经元细胞(silence-PRDX5 DIV)+氯胺酮组(silence-PRDX5 DIV+K组)及PRDX5基因过表达原代海马神经元细胞(overexpression-PRDX5 DIV)+氯胺酮组(overexpression-PRDX5 DIV+K组),细胞培养至第8天,收集4组细胞,四唑氮化合物(MTS)检测细胞增殖,原位缺口末端标记法(TUNEL)法检测细胞凋亡率,实时定量聚合酶链反应(Real-time PCR)技术检测各组细胞PRDX5下游凋亡相关基因B细胞淋巴瘤/白血病-2(bcl-2)、bcl-2相关X蛋白(bax)在mRNA转录水平。组间比较采用t检验。结果体内条件下,PRDX免疫组化显示,对照组[(72.2±0.4)%]、丙泊酚组[(55.3±0.1)%]、氯胺酮+丙泊酚组[(42.6±0.3)%]、氯胺酮组[(17.5±0.2)%],PRDX免疫组化阳性面积百分比依次减少(t=2.110,P<0.05)。体外条件下,细胞凋亡率:silence-PRDX5DIV+K组[(21.2±0.8)%]、DIV+K组[(13.3±0.5)%]、overexpression-PRDX5DIV+K组[(7.4±0.4)%]、DIV+D组[(3.2±0.3)%],细胞凋亡率依次降低(t=1.520,P<0.05)。Real-time PCR技术检测显示:海马神经元细胞凋亡相关基因bcl-2在mRNA水Objective To investigate the mechanism of Peroxiredoxin 5(PRDX5)in the reduction of ketamine-induced neuronal apoptosis by propofol.Methods Research time:From March 2018 to January 2019,80 healthy 7-day-old Sprague-Dawley(SD)male pups(Beijing Weitong Lihua Experimental Animal Company Certificate Number:218765245)were selected,and the pups were randomly divided For 4 groups(n=20),the control group was intraperitoneally injected with 0.9%saline 1 ml(control group),experimental group A was intraperitoneally injected with 80 mg/kg ketamine 1ml(ketamine group),and experimental group B was intraperitoneally injected with 80 mg/kg propofol 1ml(Propofol group),experimental group C intraperitoneal injection(80 mg/kg ketamine+80 mg/kg propofol)total 1ml(propofol+ketamine group).After the rats were awakened,they continued to feed for 3 weeks and then performed a platform jumping experiment(n=10).After the test,the hippocampus tissue was sacrificed and the PRDX5 content was detected by immunohistochemistry(n=10).Under in vitro culture conditions,clean SD rats(Beijing Weitonglihua Experimental Animal Company Certificate Number:218764758)were used to culture primary hippocampal neurons,divided into four groups:primary hippocampal neurons(DIV)+Distilled water group(DIV+D group),primary hippocampal neuron cells(DIV)+ketamine group(DIV+K group),PRDX5 gene silenced primary hippocampal neuronal cells(silence-PRDX5 DIV)+ketamine group(silence-PRDX5)DIV+K group and PRDX5 gene overexpression primary hippocampal neuron cells(overexpression-PRDX5 DIV)+ketamine group(overexpression-PRDX5DIV+K group),the cells were cultured to the 8th day,four groups of cells were collected,and MTS was used to detect cell proliferation.The TUNEL method was used to detect the rate of cell apoptosis,and the Real-time PCR technology was used to detect the mRNA transcription levels of the apoptosis-related genes B cell lymphoma/leukemia-2(bcl-2)and bcl-2 associated X protein(bax)downstream of PRDX5 in each group.The data comparison between groups is performed

关 键 词：丙泊酚 过氧化物氧化还原蛋白-5 氯胺酮 神经细胞 脱噬作用 

分 类 号：R614[医药卫生—麻醉学]