摘要
目的探讨自噬在机械通气致大鼠脑组织神经元凋亡中的作用。方法健康成年雄性Wistar大鼠24只,采用随机数字表法平均分为4组:对照组(C组)、机械通气组(MV组)、机械通气+6-氨基-3-甲基嘌呤处理组(3MA组),机械通气+雷帕霉素处理组(R组)。C组气管插管后保留自主呼吸,不行机械通气; MV组、3MA组、R组均行机械通气。3MA组及R组于机械通气前15 min分别腹腔注射自噬抑制剂3-MA 30 mg/kg及自噬激动剂雷帕霉素1 mg/kg,C组于MV前15 min腹腔注射等量生理盐水。于机械通气结束时处死大鼠取脑组织,光镜下观察病理学结果。采用TUNEL法检测海马神经元凋亡情况,计算凋亡指数;采用Western blot检测Bax、Bcl-2、活化型Caspase-3、LC3 I、LC3Ⅱ、p62的表达,并计算Bax/Bcl-2、LC3Ⅱ/Ⅰ比值。结果与C组相比,MV组、3MA组和R组病理学损伤严重,海马神经元凋亡指数和Bax/Bcl-2比值升高,海马活化型Caspase-3表达上调,Beclin-1、LC3Ⅱ/Ⅰ表达上调,p62表达下调(P <0. 05);与MV组比较,3MA组病理学损伤减轻,海马神经元凋亡指数和Bax/Bcl-2比值降低,海马活化型Caspase-3表达下调,Beclin-1、LC3Ⅱ/I表达下调,p62表达上调(P <0. 05);与MV组比较,R组大鼠海马神经元凋亡指数和Bax/Bcl-2比值升高,海马活化型Caspase-3表达上调,Beclin-1、LC3Ⅱ/I表达上调,p62表达下调(P <0. 05)。结论长时间大潮气量机械通气致大鼠脑组织神经元凋亡的机制与自噬有关。
Objective To evaluate the role of autophagy in apoptosis of neurons in brain tissue of rats induced by mechanical ventilation.Methods A total of 24 healthy adult male Wistar rats were divided into 4 groups(n=6)by random number table method,including control group(C group),mechanical ventilation group(MV group),mechanical ventilation combined with 6-amino-3-methylindole treatment group(3MA group),and mechanical ventilation plus rapamycin treatment group(R group).Group C retained spontaneous breathing after endotracheal intubation without mechanical ventilation.MV group,3MA group and R group were performed mechanical ventilation.3MA group and the R group received intraperitoneal injection of 30 mg/kg 3-MA and 1 mg/kg rapamycin,respectively,at 15 min before MV,while the C group was given intraperitoneal injection of the same amount of normal saline at 15 min before MV.At the end of mechanical ventilation,6 rats were sacrificed for brain tissue to observe pathological changes under light microscope.TUNEL assay was used to detect the apoptosis of neurons in the hippocampus CA1 region and calculate the apoptosis index.Using Western blot method to detect the expression of Bax,Bcl-2,activated Caspase-3,LC3Ⅰ,LC3Ⅱ,p62,and calculate the ratios of Bax to Bcl-2,LC3Ⅱto LC3Ⅰ.Results Compared with group C,the rest three groups had severe pathological damage,with higher hippocampus neuron apoptosis index and Bax/Bcl-2 ratio,up-regulated expression of activated Caspase-3,increased Beclin-1,LC3Ⅱ/Ⅰand decreased p62(P<0.05);compared with MV group,pathological damage,the hippocampus neuron apoptosis index and Bax/Bcl-2 ratio,the expression of activated Caspase-3,Beclin-1,LC3Ⅱ/Ⅰwas decreased,p62 was up-regulated in 3MA group(P<0.05);compared with MV group,the changes of above indicators in the R group was contrary to 3M group(P<0.05).Conclusion Autophagy is involved in the apoptosis of neurons in rat brain tissue induced by high tidal volume ventilation.
作者
余鑫
米智华
高巨
YU Xin;MI Zhihua;GAO Ju(Clinical Medical School of Yangzhou University Subei People′s Hospital of Jiangsu Province,Yangzhou,Jiangsu,225001)
出处
《实用临床医药杂志》
CAS
2019年第8期12-15,21,共5页
Journal of Clinical Medicine in Practice
基金
国家自然科学基金资助项目(81571936
81601679
81401626)
作者简介
通信作者:高巨,E-mail:gaoju_003@163.com.
