中脑腹侧被盖区多巴胺转运体在丙泊酚全麻大鼠苏醒中的作用  

The role of midbrain VTA dopamine transporter(DAT)in propofol general anesthesia

作　　者：郭佳 许珂 殷姜文 葛明月 秦新磊 樊世文 张晗 李燕[2] GUO Jia;XU Ke;YIN Jiangwen;GE Mingyue;QIN Xinlei;FAN Shiwen;ZHANG Han;LI Yan(School of Medicine,Shihezi University,Shihezi,Xinjiang 832002,China;Department of Anesthesiology,the First Affiliated Hospital,School of Medicine,Shihezi University,Shihezi,Xinjiang 832002,China)

机构地区：[1]石河子大学医学院,新疆石河子832000 [2]石河子大学医学院第一附属医院,新疆石河子832000

出　　处：《石河子大学学报（自然科学版）》2021年第2期234-239,共6页Journal of Shihezi University(Natural Science)

基　　金：国家自然科学基金项目(81860209),石河子大学医学院第一附属医院重点基金项目(ZD201705)。

摘　　要：目的研究中脑腹侧被盖区(VTA)多巴胺转运体(DAT)在丙泊酚麻醉苏醒中的作用。方法将60只大鼠随机分为正常对照组、6-OHDA毁损组、生理盐水组、丙泊酚麻醉组、AAV空载组、AAV2·DAT·RNAi组,每组10只。AAV干预大鼠预先经脑立体定位仪显微靶向注射AAV至VTA核团。各组大鼠经尾静脉以11 mg·kg^(-1)的剂量缓慢推注丙泊酚进行麻醉诱导,后以70 mg·kg^(-1)·h^(-1)的剂量持续泵入丙泊酚,对照组以相同速度输注等剂量生理盐水,共30 min,停药10 min后取脑;采用免疫荧光染色观察VTA内DAT蛋白、前额叶皮层c-fos蛋白的表达情况;Western blot法检测PFC内c-fos的蛋白含量。通过翻正反射消失(LORR)时间和翻正反射恢复(RORR)时间记录丙泊酚麻醉诱导和苏醒时间;应用在体脑电记录大鼠麻醉期和苏醒期的脑电变化。结果与NC组比较,6-OHDA组LORR时间缩短,RORR时间延长(P<0.05);与生理盐水组比较,丙泊酚麻醉组VTA内DAT表达降低(P<0.05);与NC组比较,AAV组LORR时间延长,RORR时间缩短,脑电图在丙泊酚麻醉期间α波、β波频率增多(P<0.05),在丙泊酚苏醒期间β波频率增多,θ波频率减少,丙泊酚麻醉苏醒期间PFC中c-fos蛋白表达增多(P<0.05);与NC组比较,AAV·NC组LORR时间和RORR时间、丙泊酚麻醉及苏醒期间的脑电图、丙泊酚麻醉苏醒期PFC中c-fos蛋白表达均无统计学差异(P>0.05)。结论通过敲低VTA区DAT,可使PFC神经元活性增强,促进大鼠从丙泊酚麻醉中苏醒。Objective To investigate the role of dopamine transporter(DAT)in the ventral tegmental area(VTA)in propofol anesthesia recovery.Methods 60 SD rats were randomly divided into negative control group,6-OHDA lesion group,saline group,propofol anesthesia group,AAV·NC group and AAV2·DAT·RNAi group,with 10 rats in each group.The AAV2·DAT·RNAi group was injected with AAV to VTA nuclei by microtargeting via stereotactic locator.Propofol was slowly injected into the tail vein in each group at a dose of 11 mg·kg^(-1)for anesthesia induction,then continuously pumped at a dose of 70 mg·kg^(-1)·h^(-1).The control group was injected with the same dose of normal saline at the same speed for a total of 30min,and the brain was taken after withdrawal for 10min.Immunofluorescence staining was used to observe the expression of DAT protein and c-fos protein in the prefrontal cortex in VTA.Western blot was used to detect c-fos protein in PFC.Propofol anesthesia induction and resuscitation time were recorded by LORR and RORR time.The electroencephalogram(EEG)was used to record the changes of rats during anesthesia and awakening.Results Compared with NC group,LORR time and RORR time of 6-OHDA group were shortened and prolonged(P<0.05).Compared with the saline group,DAT expression in the propofol anesthesia group was decreased(P<0.05).Compared with the NC group,the LORR time and RORR time in the AAV group were prolonged,the frequency ofαwave andβwave increased during propofol anesthesia,theβwave frequency increased during propofol recovery,theθwave frequency decreased during propofol recovery,and c-fos protein expression increased during propofol recovery(P<0.05).Compared with the NC group,there were no statistically significant differences in LORR time and RORR time,、EEG changes during propofol anesthesia and recovery and c-fos protein expression during propofol anesthesia recovery in the AAV·NC group(P>0.05).Conclusion By knocking down the DAT in the VTA region,the activity of PFC neurons was enhanced and the rats were aw

关 键 词：丙泊酚 中脑腹侧被盖区 多巴胺转运蛋白 皮层脑电 麻醉-觉醒 

分 类 号：R614.1[医药卫生—麻醉学]