中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2013年
19期
1507-1511
,共5页
蔺芳菊%杨晓苏%杨德%邹艳群
藺芳菊%楊曉囌%楊德%鄒豔群
린방국%양효소%양덕%추염군
入睡和睡眠障碍%阳离子-氯离子共转运体%KCC2%NKCC1
入睡和睡眠障礙%暘離子-氯離子共轉運體%KCC2%NKCC1
입수화수면장애%양리자-록리자공전운체%KCC2%NKCC1
Sleep initiation and maintenance disorders%Cation-chloride cotransporters%KCC2%NKCC1
目的 探讨阳离子-氯离子共转运体KCC2、NKCC1在大鼠急性失眠发生机制中的作用.方法 SD大鼠腹腔注射对氯苯丙氨酸(PCPA)制作急性失眠大鼠模型,同步设立生理盐水对照组和地西泮干预组,每组6只；用逆转录-聚合酶链反应(RT-PCR)、免疫印迹(Western blot)检测各组大鼠脑干KCC2、NKCC1的表达；用氯离子荧光探针(MQAE),结合激光共聚焦显微镜观察脑干组织细胞内氯离子浓度([Cl-]i).结果 (1)模型组和干预组的KCC2 mRNA及蛋白表达均低于对照组(mRNA丰度值:0.196±0.021比0.939 ±0.109,P＜0.05；0.485±0.026比0.939±0.109,P＜0.05；蛋白相对值:0.363 ±0.058比0.967±0.155,P＜0.05; 0.663 ±0.106比0.967 ±0.155,P＜0.05)；干预组高于模型组(mRNA丰度值:0.485±0.026比0.196±0.021,P＜0.05；蛋白相对值:0.663±0.106比0.363±0.058,P＜0.05).(2)模型组的NKCCl mRNA及蛋白表达稍高于对照组,但差异无统计学意义(mRNA丰度值:0.344±0.026比0.320±0.019,P＞0.05；蛋白相对值:0.244±0.010比0.230±0.021,P＞0.05)；干预组较对照组和模型组降低(mRNA丰度值:0.066±0.031比0.320±0.019,P＜0.05；0.066±0.031比0.344±0.026,P＜0.05；蛋白相对值:0.131±0.012比0.230±0.021,P＜0.05;0.131±0.012比0.244±0.010,P＜0.05).(3)模型组的[C1-]i高于对照组和干预组(相对值:0.0315±0.0039比0.0164±0.0019,P＜0.05;0.0315±0.0039比0.0182±0.0013,P＜0.05)；干预组稍高于对照组,但差异无统计学意义(相对值:0.0182±0.0013比0.0164±0.0019,P＞0.05).结论 (1)脑干KCC2表达下调及[Cl-]i的升高可能参与大鼠急性失眠的病理机制；(2)地西泮可能通过上调KCC2和下调NKCC1的表达,进而降低[Cl-]i而发挥镇静催眠作用.
目的 探討暘離子-氯離子共轉運體KCC2、NKCC1在大鼠急性失眠髮生機製中的作用.方法 SD大鼠腹腔註射對氯苯丙氨痠(PCPA)製作急性失眠大鼠模型,同步設立生理鹽水對照組和地西泮榦預組,每組6隻；用逆轉錄-聚閤酶鏈反應(RT-PCR)、免疫印跡(Western blot)檢測各組大鼠腦榦KCC2、NKCC1的錶達；用氯離子熒光探針(MQAE),結閤激光共聚焦顯微鏡觀察腦榦組織細胞內氯離子濃度([Cl-]i).結果 (1)模型組和榦預組的KCC2 mRNA及蛋白錶達均低于對照組(mRNA豐度值:0.196±0.021比0.939 ±0.109,P＜0.05；0.485±0.026比0.939±0.109,P＜0.05；蛋白相對值:0.363 ±0.058比0.967±0.155,P＜0.05; 0.663 ±0.106比0.967 ±0.155,P＜0.05)；榦預組高于模型組(mRNA豐度值:0.485±0.026比0.196±0.021,P＜0.05；蛋白相對值:0.663±0.106比0.363±0.058,P＜0.05).(2)模型組的NKCCl mRNA及蛋白錶達稍高于對照組,但差異無統計學意義(mRNA豐度值:0.344±0.026比0.320±0.019,P＞0.05；蛋白相對值:0.244±0.010比0.230±0.021,P＞0.05)；榦預組較對照組和模型組降低(mRNA豐度值:0.066±0.031比0.320±0.019,P＜0.05；0.066±0.031比0.344±0.026,P＜0.05；蛋白相對值:0.131±0.012比0.230±0.021,P＜0.05;0.131±0.012比0.244±0.010,P＜0.05).(3)模型組的[C1-]i高于對照組和榦預組(相對值:0.0315±0.0039比0.0164±0.0019,P＜0.05;0.0315±0.0039比0.0182±0.0013,P＜0.05)；榦預組稍高于對照組,但差異無統計學意義(相對值:0.0182±0.0013比0.0164±0.0019,P＞0.05).結論 (1)腦榦KCC2錶達下調及[Cl-]i的升高可能參與大鼠急性失眠的病理機製；(2)地西泮可能通過上調KCC2和下調NKCC1的錶達,進而降低[Cl-]i而髮揮鎮靜催眠作用.
목적 탐토양리자-록리자공전운체KCC2、NKCC1재대서급성실면발생궤제중적작용.방법 SD대서복강주사대록분병안산(PCPA)제작급성실면대서모형,동보설립생리염수대조조화지서반간예조,매조6지；용역전록-취합매련반응(RT-PCR)、면역인적(Western blot)검측각조대서뇌간KCC2、NKCC1적표체；용록리자형광탐침(MQAE),결합격광공취초현미경관찰뇌간조직세포내록리자농도([Cl-]i).결과 (1)모형조화간예조적KCC2 mRNA급단백표체균저우대조조(mRNA봉도치:0.196±0.021비0.939 ±0.109,P＜0.05；0.485±0.026비0.939±0.109,P＜0.05；단백상대치:0.363 ±0.058비0.967±0.155,P＜0.05; 0.663 ±0.106비0.967 ±0.155,P＜0.05)；간예조고우모형조(mRNA봉도치:0.485±0.026비0.196±0.021,P＜0.05；단백상대치:0.663±0.106비0.363±0.058,P＜0.05).(2)모형조적NKCCl mRNA급단백표체초고우대조조,단차이무통계학의의(mRNA봉도치:0.344±0.026비0.320±0.019,P＞0.05；단백상대치:0.244±0.010비0.230±0.021,P＞0.05)；간예조교대조조화모형조강저(mRNA봉도치:0.066±0.031비0.320±0.019,P＜0.05；0.066±0.031비0.344±0.026,P＜0.05；단백상대치:0.131±0.012비0.230±0.021,P＜0.05;0.131±0.012비0.244±0.010,P＜0.05).(3)모형조적[C1-]i고우대조조화간예조(상대치:0.0315±0.0039비0.0164±0.0019,P＜0.05;0.0315±0.0039비0.0182±0.0013,P＜0.05)；간예조초고우대조조,단차이무통계학의의(상대치:0.0182±0.0013비0.0164±0.0019,P＞0.05).결론 (1)뇌간KCC2표체하조급[Cl-]i적승고가능삼여대서급성실면적병리궤제；(2)지서반가능통과상조KCC2화하조NKCC1적표체,진이강저[Cl-]i이발휘진정최면작용.
Objective To explore the possible roles of KCC2 and NKCC1 in the pathological mechanism of acute insomnia in rats.Methods A total of 18 Sprague-Dawley rats were randomly selected into model,interference and normal control groups.The expressions of KCC2 and NKCC1 in brainstem were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot.The concentration of intracellular C1-([Cl-] i) in brainstem was detected by fluorescence probe MQAE with laser confocal microscopy.Results (1) Comparing with the control group,both KCC2 mRNA and protein expression were down-regulated in the model and interference groups (mRNA:0.196 ±0.021 vs 0.939 ±0.109,P ＜0.05;0.485 ±0.026 vs 0.939 ±0.109,P ＜0.05; protein expression:0.363 ± 0.058 vs 0.967 ± 0.155,P ＜0.05 ; 0.663 ± 0.106 vs 0.967 ± 0.155,P ＜ 0.05).However they became up-regulated in the interference group versus the model group (mRNA:0.485 ± 0.026 vs 0.196 ± 0.021,P ＜ 0.05 ; protein expression:0.663 ±0.106 vs 0.363 ±0.058,P ＜0.05).(2) Comparing with the control group,both NKCCI mRNA and protein expression in the model group were slightly up-regulated.But statistical difference was insignificant (mRNA:0.344 ± 0.026 vs 0.320 ± 0.019,P ＞ 0.05 ; protein expression:0.244 ± 0.010 vs0.230 ± 0.021,P ＞ 0.05).There was down-regulation in the interference group versus the model and control groups (mRNA:0.066 ± 0.031 vs 0.320 ± 0.019,P ＜ 0.05 ; 0.066 ± 0.031 vs 0.344 ± 0.026,P ＜ 0.05 ;protein expression:0.131 ± 0.012 vs 0.230 ± 0.021,P ＜ 0.05 ; 0.131 ± 0.012 vs 0.244 ± 0.010,P ＜0.05).(3) Comparing with the control group,[C1-]i became up-regulated in the model group (0.0315 ±0.0039 vs 0.0164 ±0.0019,P ＜0.05).It was down-regulated in the interference group versus the model group (0.0182 ±0.0013 vs 0.0315 ±0.0039,P ＜0.05),but higher than control group without statistical difference (0.0182 ±0.0013 vs 0.0164 ±0.0019,P ＞0.05).Conclusion The down-regulation of KCC2 and rise of [C1-] i in brainstem may participate in the pathological mechanism of acute insomnia in rats.And the mechanism of sedative-hypnotic diazepam may be operate through an up-regulation of KCC2,a down-regulation of NKCC1 and decreased [C1-] i.
