缝隙连接蛋白43在卒中后抑郁模型大鼠海马的表达及其对细胞凋亡的影响  

Expression of connexin 43 in hippocampus of post-stroke depression model rats and its effect on cell apoptosis

作　　者：陈金梅 王喻 王洁[3] 罗雪莲 王功俊 包成政 李雪斌[1,2] Chen Jinmei;Wang Yu;Wang Jie;Luo Xuelian;Wang Gongjun;Bao Chengzheng;Li Xuebin(Youjiang Medical University for Nationalities,Baise 533000,China;Key Laboratory of the West Guangxi Endemic Diseases Prevention and Treatment in Guangxi Province,Baise 533000,China;Department of Nephrology,Affiliated Hospital of Youjiang Medical University for Nationalities,Baise 533000,China)

机构地区：[1]右江民族医学院,百色533000 [2]广西高校桂西地区高发病防治研究重点实验室,百色533000 [3]右江民族医学院附属医院肾内科,百色533000

出　　处：《中华行为医学与脑科学杂志》2024年第6期487-493,共7页Chinese Journal of Behavioral Medicine and Brain Science

基　　金：国家自然科学基金(81860226);广西壮族自治区卫生和计划生育委员会中医药科技专项(GZLC16);广西医疗卫生适宜技术开发与推广应用项目(S2018073);广西研究生教育创新计划项目(YCSW2021329)。

摘　　要：目的探讨缝隙连接蛋白43(connexin 43,Cx43)在卒中后抑郁(post-stroke depression,PSD)大鼠海马的表达及其对细胞凋亡及抑郁样行为的影响。方法60只6~8周龄清洁级雄性SD大鼠,随机分为5组(每组12只):正常组、卒中组、抑郁组、PSD组、甘珀酸(carbenoxolone,CBX)组。采用内皮素-1注射法建立卒中模型,慢性不可预见的温和性应激(chronic unpredictable mild stress,CUMS)结合孤养建立抑郁模型。PSD组大鼠在卒中造模第7天给予CUMS及孤养。CBX组在PSD造模第14天给予腹腔注射CBX(20 mg/kg)。采用糖水偏爱实验、旷场实验评价大鼠抑郁行为,RT-PCR检测大鼠组织海马Cx43 mRNA表达,Western blot检测Cx43、caspase-3、Bax、Bcl-2的蛋白表达水平,TUNEL染色检测细胞凋亡变化。采用SPSS 23.0软件进行统计分析,行为学数据采用重复测量的方差分析,其他数据多组间采用单因素方差分析,两两比较采用LSD-t检验。结果(1)5组大鼠糖水偏爱率及穿格次数比较,时间和组别的交互作用均显著(F_(交互)=35.57,111.43,均P<0.05)。在术后28 d,抑郁组、PSD组糖水偏爱率及穿格次数均低于卒中组(均P<0.05),CBX组糖水偏爱率、穿格次数均低于PSD组(均P<0.05)。(2)5组大鼠Cx43 mRNA、Cx43蛋白水平比较均差异有统计学意义(F=273.57,64.56,均P<0.05)。抑郁组[(0.59±0.05),(0.69±0.08)]与PSD组[(0.61±0.07),(0.63±0.12)]Cx43 mRNA及CX43蛋白水平均低于卒中组[(1.01±0.03),(1.05±0.08)](均P<0.05)。CBX组Cx43 mRNA及Cx43蛋白水平[(0.30±0.01),(0.37±0.09)]均低于PSD组(均P<0.05)。(3)5组大鼠caspase-3、Bax、Bcl-2的蛋白水平及Bcl-2/Bax、细胞凋亡率均差异均有统计学意义(F=102.40,90.27,47.42,159.99,115.21,均P<0.05)。卒中组[(0.44±0.06),(0.54±0.07),(29.16±5.03)]与抑郁组[(0.45±0.07),(0.59±0.09),(27.00±4.93)]caspase-3、Bax蛋白水平、细胞凋亡率均高于正常组[(0.21±0.08),(0.33±0.07),(4.83±3.18)](均P<0.05),卒中组[(0.80±0.04),(1.51±0.20)]与抑郁组[Objective To explore the expression of connexin 43(Cx43)in hippocampus of post-stroke depression(PSD)model rats and its effect on cell apoptosis and depressive-like behavior.Methods Sixty SPF-grade male SD rats aged 6-8 weeks were randomly divided into five groups(12 rats in each group):normal group,stroke group,depression group,PSD group and carbenoxolone(CBX)group.The stroke model was established by injection of endothelin-1.Chronic unpredictable mild stress(CUMS)combined with solitary rearing was used to establish a depression model.Rats in PSD group were given CUMS and raised alone on the seventh day of stroke modeling.Rats in CBX group were given intraperitoneal injection of CBX(20 mg/kg)on 14th day after PSD modeling.The depressive-like behavior of rats was evaluated by sugar water preference test and open field test.The expression of Cx43 mRNA in hippocampus of rats was detected by RT-PCR,the expression levels of Cx43,caspase-3,Bax and Bcl-2 were detected by Western blot,and the changes of apoptosis rate were detected by TUNEL staining.SPSS 23.0 software was used for statistical analysis,the behavioral data were analyzed by repeated measurement ANOVA,the remaining data were analyzed by one-way ANOVA,and the LSD-t test was used for further pairwise comparison.Results(1)As for the preference rate of sugar water and the times of crossing the grid,the interaction effects between time and group were significant among the 5 groups(Finteraction=35.57,111.43,both P<0.05).On the 28th day after operation,the preference rate of sugar water and the times of crossing grid in depression group and PSD group were lower than those in stroke group(all P<0.05),while the preference rate of sugar water and the times of crossing grid in CBX group were both lower than those in PSD group(both P<0.05).(2)The levels of Cx43 mRNA and Cx43 protein in the five groups were significantly different(F=273.57,64.56,both P<0.05).The levels of Cx43 mRNA and Cx43 protein in depression group((0.59±0.05),(0.69±0.08))and PSD group((0.61±0.07)

关 键 词：卒中后抑郁 缝隙连接蛋白43 凋亡 海马 大鼠 

分 类 号：R743.3[医药卫生—神经病学与精神病学] R-332[医药卫生—临床医学]