首页|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

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

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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SPF级雄性SD大鼠,按随机数字表法分为5组(每组12只):正常组、卒中组、抑郁组、PSD组、甘珀酸(carbenoxolo-ne,CBX)组.采用内皮素-1注射法建立卒中模型,慢性不可预见性温和性应激(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方差分析,其他数据多组间比较采用单因素方差分析,进一步两两比较采用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[(0.60±0.09),(1.03±0.09)]Bcl-2 蛋白水平及 Bcl-2/Bax均低于正常组[(1.04±0.13),(3.14±0.38)](均 P<0.05).PSD 组 caspase-3、Bax 蛋白水平、细胞凋亡率[(0.76±0.05),(0.84±0.02),(44.50±3.83)]均高于卒中组与抑郁组(均 P<0.05),PSD 组Bcl-2 蛋白水平、Bcl-2/Bax[(0.50±0.14),(0.59±0.17)]均低于卒中组与抑郁组(均 P<0.05).CBX组 caspase-3、Bax 蛋白水平、细胞凋亡率[(1.03±0.10),(1.02±0.05),(56.00±4.81)]均高于 PSD 组(均 P<0.05).CBX 组 Bcl-2 蛋白水平、Bcl-2/Bax[(0.26±0.08),(0.25±0.08)]均低于 PSD 组(均 P<0.05).结论 PSD模型大鼠海马Cx43表达下调,可促进细胞凋亡发生,进而加重抑郁行为.
Expression of connexin 43 in hippocampus of post-stroke depression model rats and its effect on cell apoptosis
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.Meth-ods Sixty SPF-grade male SD rats aged 6-8 weeks were randomly divided into five groups according to the random number table method(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.Chro-nic 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 de-pressive-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 wa-ter 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),(0.63±0.12))were lower than those in stroke group((1.01±0.03),(1.05±0.08))(all P<0.05).The levels of Cx43 mRNA and Cx43 protein in CBX group((0.30±0.01),(0.37±0.09))were lower than those in PSD group(both P<0.05).(3)The protein levels of caspase-3,Bax,Bcl-2 and Bcl-2/Bax and the apopto-sis rate of the five groups were significantly different(F=102.40,90.27,47.42,159.99,115.21,all P<0.05).The levels of caspase-3,Bax protein,apoptosis rate in stroke group((0.44±0.06),(0.54±0.07),(29.16±5.03))and depression group((0.45±0.07),(0.59±0.09),(27.00±4.93))were higher than those in normal group((0.21±0.08),(0.33±0.07),(4.83±3.18))(all P<0.05),the levels of Bcl-2 protein and Bcl-2/Bax in stroke group((0.80±0.04),(1.51±0.20))and depression group((0.60±0.09),(1.03±0.09))were lower than those in normal group((1.04±0.13),(3.14±0.38))(all P<0.05).The levels of caspase-3,Bax protein and apoptosis rate in PSD group((0.76±0.05),(0.84±0.02),(44.50±3.83))were all higher than those in stroke group and depression group(all P<0.05),and the levels of Bcl-2 protein and Bcl-2/Bax in PSD group((0.50±0.14),(0.59±0.17))were lower than those in stroke group and depression group(both P<0.05).The levels of caspase-3 and Bax protein and the apoptosis rate in CBX group((1.03±0.10),(1.02±0.05),(56.00±4.81))were higher than those in PSD group(all P<0.05).The levels of Bcl-2 protein and Bcl-2/Bax in CBX group((0.26±0.08),(0.25±0.08))were lower than those in PSD group(both P<0.05).Conclusion The expression level of Cx43 in the hippocampus of PSD model rats is downregulated,which can promote cell apoptosis and exacerbate depressive behavior.

Post-stroke depressionConnexin 43ApoptosisHippocampusRat

陈金梅、王喻、王洁、罗雪莲、王功俊、包成政、李雪斌

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右江民族医学院,百色 533000

右江民族医学院附属医院肾内科,百色 533000

广西高校桂西地区高发病防治研究重点实验室,百色 533000

卒中后抑郁 缝隙连接蛋白43 凋亡 海马 大鼠

国家自然科学基金广西壮族自治区卫生和计划生育委员会中医药科技专项广西医疗卫生适宜技术开发与推广应用项目广西研究生教育创新计划项目

81860226GZLC16S2018073YCSW2021329

2024

中华行为医学与脑科学杂志
中华医学会 济宁医学院

中华行为医学与脑科学杂志

CSTPCD北大核心
影响因子:1.472
ISSN:1674-6554
年,卷(期):2024.33(6)