中华预防医学杂志
中華預防醫學雜誌
중화예방의학잡지
CHINESE JOURNAL OF
2012年
7期
635-639
,共5页
姚晶%沈新南%沈慧%吴岷
姚晶%瀋新南%瀋慧%吳岷
요정%침신남%침혜%오민
大鼠%再灌注损伤%受体,谷氨酸%单胺类神经递质%磷脂酶C-γ1%茶氨酸
大鼠%再灌註損傷%受體,穀氨痠%單胺類神經遞質%燐脂酶C-γ1%茶氨痠
대서%재관주손상%수체,곡안산%단알류신경체질%린지매C-γ1%다안산
Rats%Reperfusion injury%Receptor glutamate%Monoamines%PLC-γ1%Theanine
目的 探讨茶氨酸对大鼠脑缺血再灌注后多巴胺(DA)、5-羟色胺(5-HT)的水平及对谷氨酸受体2(GluR2) mRNA、麟脂酶C-γ1( PLC-γ1) mRNA表达的影响,讨论其所致神经损伤的保护作用机制.方法 将56只6周龄、体重为100～ 120 g的SPF级雄性SD大鼠按体重分层,以随机势字表法分为5组,分别为模型组(12只)、假手术组(8只),以及茶氨酸低( 10 mg/kg)、中(30 mg/kg)、高剂量组(90 mg/kg)(每组12只).茶氨酸剂量组大鼠灌胃每天给予相应剂量的茶氨酸,模型组和假手术组每天灌胃给予等量超纯水.连续灌胃15 d后,制备大鼠大脑中动脉栓塞(middle cerebraartery occlusion,MCAO)模型,于再灌注后第3、24小时进行神经学评分,再灌注后第24小时处死动物,测定脑组织中DA、5-HT和茶氨酸含量,同时测定脑组织线粒体中活性氧(ROS)生成和过氧化氢酶( CAT)活性,并采用RT-PCR法检测脑组织中GluR2 mRNA以及PLC-γ1 mRNA的表达水平.结果 模型组、茶氨酸低、中、高剂量组大鼠脑缺血再灌注后第3小时神经学评分分别为(6.000±0.926)、(4.100±0.738)、(3.444±0.726)、(2.250±0.886)分(F=29.70,P＜0.01)；第24小时神经学评分分别为(6.625±0.916)、(5.000±0.817)、(3.667±0.707)、(2.625±0.916)分(F=34.68,P＜0.01).模型组、茶氨酸低、中、高剂量组及假手术组大鼠脑内DA含量分别为(10.26±1.12)、(12.48±1.09)、(14.55±0.94)、(15.97±0.92)、(11.98±0.63) μg/g(F=43.76,P＜0.01);5-HT含量分别为(1.091±0.160)、(0.818±0.101)、(0.571±0.050)、(0.453±0.111)、(0.863±0.063) μg/g(F =48.68,P＜0.01)；大鼠脑线粒体中ROS生成速率分别为(3.072±0.503)、(1.331±0.268)、(1.295±0.061)、(0.804±0.200)、(2.158±0.218)U· min-1· mg-1(F =80.82,P ＜0.01);CAT活性分别为(4.880±1.121)、(8.405±1.356)、(9.535±2.511)、(15.090±4.054)、(21.260±6.054) U/g(F =28.58,P＜0.01)；大鼠脑内GluR2 mRNA相对表达量分别为0.842±0.020、1.063±0.100、1.170±0.152、1.254±0.131、1.012±0.056(F=9.23,P＜0.01); PLC-γ1mRNA相对表达量分别为0.737±0.090、0.887±0.045、0.963±0.025、0.991±0.049、0.867±0.079(F=10.24,P＜0.01).结论 茶氨酸对大鼠脑缺血再灌注损伤具有一定的保护作用,其机制与调节脑内单胺类神经递质含量以及上调GluR2 mRNA和PLC-γ1 mRNA表达有关.
目的 探討茶氨痠對大鼠腦缺血再灌註後多巴胺(DA)、5-羥色胺(5-HT)的水平及對穀氨痠受體2(GluR2) mRNA、麟脂酶C-γ1( PLC-γ1) mRNA錶達的影響,討論其所緻神經損傷的保護作用機製.方法 將56隻6週齡、體重為100～ 120 g的SPF級雄性SD大鼠按體重分層,以隨機勢字錶法分為5組,分彆為模型組(12隻)、假手術組(8隻),以及茶氨痠低( 10 mg/kg)、中(30 mg/kg)、高劑量組(90 mg/kg)(每組12隻).茶氨痠劑量組大鼠灌胃每天給予相應劑量的茶氨痠,模型組和假手術組每天灌胃給予等量超純水.連續灌胃15 d後,製備大鼠大腦中動脈栓塞(middle cerebraartery occlusion,MCAO)模型,于再灌註後第3、24小時進行神經學評分,再灌註後第24小時處死動物,測定腦組織中DA、5-HT和茶氨痠含量,同時測定腦組織線粒體中活性氧(ROS)生成和過氧化氫酶( CAT)活性,併採用RT-PCR法檢測腦組織中GluR2 mRNA以及PLC-γ1 mRNA的錶達水平.結果 模型組、茶氨痠低、中、高劑量組大鼠腦缺血再灌註後第3小時神經學評分分彆為(6.000±0.926)、(4.100±0.738)、(3.444±0.726)、(2.250±0.886)分(F=29.70,P＜0.01)；第24小時神經學評分分彆為(6.625±0.916)、(5.000±0.817)、(3.667±0.707)、(2.625±0.916)分(F=34.68,P＜0.01).模型組、茶氨痠低、中、高劑量組及假手術組大鼠腦內DA含量分彆為(10.26±1.12)、(12.48±1.09)、(14.55±0.94)、(15.97±0.92)、(11.98±0.63) μg/g(F=43.76,P＜0.01);5-HT含量分彆為(1.091±0.160)、(0.818±0.101)、(0.571±0.050)、(0.453±0.111)、(0.863±0.063) μg/g(F =48.68,P＜0.01)；大鼠腦線粒體中ROS生成速率分彆為(3.072±0.503)、(1.331±0.268)、(1.295±0.061)、(0.804±0.200)、(2.158±0.218)U· min-1· mg-1(F =80.82,P ＜0.01);CAT活性分彆為(4.880±1.121)、(8.405±1.356)、(9.535±2.511)、(15.090±4.054)、(21.260±6.054) U/g(F =28.58,P＜0.01)；大鼠腦內GluR2 mRNA相對錶達量分彆為0.842±0.020、1.063±0.100、1.170±0.152、1.254±0.131、1.012±0.056(F=9.23,P＜0.01); PLC-γ1mRNA相對錶達量分彆為0.737±0.090、0.887±0.045、0.963±0.025、0.991±0.049、0.867±0.079(F=10.24,P＜0.01).結論 茶氨痠對大鼠腦缺血再灌註損傷具有一定的保護作用,其機製與調節腦內單胺類神經遞質含量以及上調GluR2 mRNA和PLC-γ1 mRNA錶達有關.
목적 탐토다안산대대서뇌결혈재관주후다파알(DA)、5-간색알(5-HT)적수평급대곡안산수체2(GluR2) mRNA、린지매C-γ1( PLC-γ1) mRNA표체적영향,토론기소치신경손상적보호작용궤제.방법 장56지6주령、체중위100～ 120 g적SPF급웅성SD대서안체중분층,이수궤세자표법분위5조,분별위모형조(12지)、가수술조(8지),이급다안산저( 10 mg/kg)、중(30 mg/kg)、고제량조(90 mg/kg)(매조12지).다안산제량조대서관위매천급여상응제량적다안산,모형조화가수술조매천관위급여등량초순수.련속관위15 d후,제비대서대뇌중동맥전새(middle cerebraartery occlusion,MCAO)모형,우재관주후제3、24소시진행신경학평분,재관주후제24소시처사동물,측정뇌조직중DA、5-HT화다안산함량,동시측정뇌조직선립체중활성양(ROS)생성화과양화경매( CAT)활성,병채용RT-PCR법검측뇌조직중GluR2 mRNA이급PLC-γ1 mRNA적표체수평.결과 모형조、다안산저、중、고제량조대서뇌결혈재관주후제3소시신경학평분분별위(6.000±0.926)、(4.100±0.738)、(3.444±0.726)、(2.250±0.886)분(F=29.70,P＜0.01)；제24소시신경학평분분별위(6.625±0.916)、(5.000±0.817)、(3.667±0.707)、(2.625±0.916)분(F=34.68,P＜0.01).모형조、다안산저、중、고제량조급가수술조대서뇌내DA함량분별위(10.26±1.12)、(12.48±1.09)、(14.55±0.94)、(15.97±0.92)、(11.98±0.63) μg/g(F=43.76,P＜0.01);5-HT함량분별위(1.091±0.160)、(0.818±0.101)、(0.571±0.050)、(0.453±0.111)、(0.863±0.063) μg/g(F =48.68,P＜0.01)；대서뇌선립체중ROS생성속솔분별위(3.072±0.503)、(1.331±0.268)、(1.295±0.061)、(0.804±0.200)、(2.158±0.218)U· min-1· mg-1(F =80.82,P ＜0.01);CAT활성분별위(4.880±1.121)、(8.405±1.356)、(9.535±2.511)、(15.090±4.054)、(21.260±6.054) U/g(F =28.58,P＜0.01)；대서뇌내GluR2 mRNA상대표체량분별위0.842±0.020、1.063±0.100、1.170±0.152、1.254±0.131、1.012±0.056(F=9.23,P＜0.01); PLC-γ1mRNA상대표체량분별위0.737±0.090、0.887±0.045、0.963±0.025、0.991±0.049、0.867±0.079(F=10.24,P＜0.01).결론 다안산대대서뇌결혈재관주손상구유일정적보호작용,기궤제여조절뇌내단알류신경체질함량이급상조GluR2 mRNA화PLC-γ1 mRNA표체유관.
Objective To study the effects of theanine on dopamine ( DA),5-hydroxy tryptamine (5-TH) and glutamate receptor 2 ( GluR2 ) mRNA,phospholipase-γ1 ( PLC-γ1 ) mRNA in cerebral ischemia-reperfusion injury rats and explore the mechanism of protective effects of theanine on the induced brain injury by ischemia-reperfusion in rats.Methods According to random number table,a total of 56 sprague-dawley rats in SPF grade about six-week old and 100 - 120 grams weighting were divided into five groups according to the body weight levels:model group ( n =12),sham-operation group ( n =8 ),low theanine group( 10 mg/kg),middle theanine group(30 mg/kg) and high theanine group(90 mg/kg).There were 12 rats in each of the theanine group.The rats in model group and sham-operation groups were given distilled water,and the rats in theanine groups were given corresponding theanine solution intragastrically for fifteen days.Then the cerebral ischemia-reperfusion injury was established by middle cerebral artery occlusion ( MCAO ).The score of neurological behavior was evaluated at the 3rd and 24th hours after reperfusion.Rats were sacrificed at 24 hours after reperfusion,the concentrations of DA,5-HT and theanine in rats brain following ischemia-reperfusion were determined.At the same time,we determined the levels of reactive oxygen species(ROS) and activities of catalase(CAT) in mitochondria of brain.The expressions of GluR2 mRNA and PLC-γ1 mRNA in rat brain were examined by reverse transcription polymerase chain reaction (RT-PCR) technique.Results The score of neurological behavior of rats in model group,theanine-low,middle,high dose groups at the 3rd hour was 6.000 ± 0.926,4.100 ± 0.738,3.444 ± 0.726and 2.250±0.886 respectively( F =29.70,P ＜ 0.01 ),and the score at the 24th hour in these groups was 6.625 ±0.916,5.000 ±0.817,3.667 ±0.707 and 2.625 ± 0.916 respectively ( F =34.68,P ＜0.01 ).The concentration of DA in model group,theanine-low,middle,high dose groups and sham-operation group was (10.26± 1.12),(12.48 ± 1.09),(14.55 ±0.94),(15.97 ±0.92) and (11.98 ± 0.63) μg/grespectively( F =43.76,P ＜ 0.01 ).The concentration of 5-HT in these groups was ( 1.091 ± 0.160 ),(0.818±0.101),(0.571 ± 0.050),(0.453 ±0.111 ) and (0.863 ±0.063) μg/g respectively (F =48.68,P ＜ 0.01 ).The level of ROS was ( 3.072 ± 0.503 ),( 1.331 ± 0.268 ),( 1.295 ± 0.061 ),( 0.804 ±0.200)and(2.158 ±0.218)U · min-1 ·mg-1 (F=80.82,P＜0.01 )respectively and the activities of CAT in these groups were ( 4.880 ± 1.121 ),( 8.405 ± 1.356 ),( 9.535 ± 2.511 ),( 15.090 ± 4.054 ) and (21.260 ± 6.054 )U/g respectively( F =28.58,P ＜ 0.01 ).The expressions of GluR2 mRNA were 0.842 ±0.020,1.063 ± 0.100,1.170 ± 0.152,1.254 ± 0.131 and 1.012 ± 0.056 respectively ( F =9.23,P ＜0.01 ).The expressions of PLC-γ1 mRNA in these groups were 0.737 ±0.090,0.887 ±0.045,0.963 ±0.025,0.991 ±0.049 and 0.867 ±0.079 respectively( F =10.24,P ＜0.01 ).Conclusion Theanine has a protective effect on the induced brain injury by ischemia-reperfusion in rats,which might be associated with its interaction with monoamine neurotransmitters and up-regulating the expressions of GluR2 mRNA and PLC-γ1 mRNA.
