羥丁酸鈉對(duì)海馬神經(jīng)細(xì)胞鈣激活鉀通道的作用_百度文庫(kù)

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4、滿霉素治療淋病性,衣原體性和雙重感染性尿道炎35例。臨床皮膚科雜志 1995;24(5 303 304。16.樊尚榮、孟蓮編譯;性傳播疾病的治療。中華婦產(chǎn)科雜志 1997;32(2 126(1997 06 16收稿麻醉科羥丁酸鈉對(duì)海馬神經(jīng)細(xì)胞鈣激活鉀通道的作用王儒蓉 王泉云 魏蔚華西醫(yī)科大學(xué)附屬第一醫(yī)院麻醉科摘要:應(yīng)用膜片鉗技術(shù)研究了羥丁酸鈉對(duì)海馬神經(jīng)元中電導(dǎo)鈣激活鉀通道的作用。實(shí)驗(yàn)選用培養(yǎng)37天的錐體神經(jīng)元,在對(duì)稱性高鉀溶液中,應(yīng)用細(xì)胞貼附式膜片和內(nèi)面向外式膜片記錄通道電流。結(jié)果發(fā)現(xiàn):(1該通道電導(dǎo)為65pS左右,對(duì)胞內(nèi) Ca2+ 1敏感,特異鉀通道阻斷劑(T EA能抑制或阻斷該通道活動(dòng);(

5、2在細(xì)胞貼附式膜片上,羥丁酸鈉明顯增加該通道的開(kāi)放概率。因此,羥丁酸鈉對(duì)海馬神經(jīng)細(xì)胞中電導(dǎo)鈣激活鉀通道具有明顯的激活作用。關(guān)鍵詞:羥丁酸鈉 鈣激活鉀通道應(yīng)用膜片鉗的細(xì)胞貼附式和內(nèi)面向外式膜片研究了羥丁酸鈉對(duì)新生SD大鼠海馬神經(jīng)元鈣激活鉀通道的作用。1 材料和方法選用新生13天SD大鼠,取海馬按照席新正等人的方法 1 進(jìn)行原代培養(yǎng)。將培養(yǎng)37天的形態(tài)典型的錐體神經(jīng)元置于對(duì)稱性高鉀溶液中,在所需要的膜片形成后先記錄下擬實(shí)驗(yàn)電壓下的單通道電流作為對(duì)照,然后分別加入1mM、2mM的羥丁酸鈉,2分鐘后,用pClamp軟件進(jìn)行單通道電流的記錄與分析 2 。統(tǒng)計(jì)分析用配對(duì)t檢驗(yàn),以p 0 05為有顯著性差異

6、。2 結(jié)果2.1 在海馬神經(jīng)元上,記錄到一種對(duì)胞內(nèi)Ca2+敏感、并且可以被特異性鉀通道阻斷劑四乙胺(T EA阻斷、具有電壓依賴性的單通道電流。由其電流 電壓( 曲線斜率計(jì)算出單通道電導(dǎo)為65 12pS。在4個(gè)膜片上觀察了羥丁酸鈉對(duì)通道動(dòng)力學(xué)特征的影響。2.2 在細(xì)胞貼附式膜片上,羥丁酸鈉明顯增加該通道的開(kāi)放概率。各給藥組較對(duì)照組均有極顯著增加482華西醫(yī)學(xué)1997;12(4 CN51-1356/R 國(guó)家級(jí)(P <0 01,2mM 組較1mM 組增加極顯著(P <0 01;而藥物對(duì)通道的電流幅值無(wú)顯著影響(P >0 05;對(duì)其開(kāi)放和關(guān)閉時(shí)間進(jìn)行指數(shù)擬合,開(kāi)放時(shí)間用單指數(shù)就能很好

7、擬合,而關(guān)閉時(shí)間需用雙指數(shù)才能較好擬合,藥物使開(kāi)放時(shí)間常數(shù)增加,慢關(guān)閉時(shí)間常數(shù)減小(P <0 05,快關(guān)閉時(shí)間常數(shù)無(wú)明顯變化(P>0 05。3 討論鈣激活鉀通道為鉀通道的重要亞型,在調(diào)節(jié)神經(jīng)細(xì)胞的興奮中起非常重要的作用 3 ,按照在對(duì)稱性高鉀(140mM 溶液中的電導(dǎo)值分為大電導(dǎo)(>150pS、中電導(dǎo)(18120pS、小電導(dǎo)(1014pS 三類 4 。本實(shí)驗(yàn)通道電導(dǎo)值為65pS 左右,屬于中電導(dǎo)(IK的鈣激活鉀通道。它可能與神經(jīng)細(xì)胞的復(fù)極化有關(guān)。羥丁酸鈉對(duì)海馬神經(jīng)細(xì)胞IK 通道的激活作用主要表現(xiàn)在增加通道的開(kāi)放概率,不影響通道的電導(dǎo)值。由于通道開(kāi)放時(shí)間只需用單指數(shù)擬合,而關(guān)閉

8、時(shí)間需用雙指數(shù)擬合,因而通道有一個(gè)開(kāi)放狀態(tài)和兩個(gè)關(guān)閉狀態(tài),藥物使通道開(kāi)放加快,慢關(guān)閉減慢。由此可見(jiàn),羥丁酸鈉影響通道動(dòng)力學(xué)過(guò)程。因此,羥丁酸鈉對(duì)海馬神經(jīng)細(xì)胞IK 通道具有明顯的激活作用。4 參考文獻(xiàn)1.席新正,葉江鴻,陳助華等:大腦皮層神經(jīng)元鉀離子通道的膜片鉗研究。中國(guó)科學(xué)B 輯,1992;10 1063。2.Hamill OP,M arty A,Neher E,et al:Im proved patchclamp techniques for high res oluti on curent recordi ng from cells and cell free membrane patch

9、es.Pflugers Arch ,1981,391:85.3.Wann KJ:Neuronal sodium and potassium chann els:structure and function.Br J Anaesth,1993,71 2.4.Blatz AL,M egleby KL:Calcium activated potassium channel.TINS,1987,10 463。(1997-09-02收稿The Effect of Sodiu m Gamma hydroxybutyrate onHippocampal Neuronal Calciu m activated

10、 PotassiumW ang Ru ron g,W ang Quanyun,W ei W eiFirst University Hospital,W est China University of Med ical Science s ,Chengd u ,610041T he purpose of this r esearch was to study the effect of sodium gamma hydrox ybutyrate on the middle conductance Calcium activated potassium (IKchannel in the cult

11、ure of hippocampal neurons of newbon rat.A ll the ex periments w ere carried out on 37days old neur ons.Single channel current signals were r ecorded in symmetrical hig h potassium w as solution under cell attached patch and inside out parch.T he results showed that:(1In inside out patch,the uni tar

12、y conductance w as 65+12pS.T he IK channel activity was activ ated by internal calcium was but w as blocked by potas sium channel blocker (T EA .(2I n cell attached patch,when sodium gamma hydrox ybutyrate w as added into thebath solution,the open probabilit y (Powas markedly increased (P<0 05,where as ther e w as no chang e in ampli tude (P>0 05。So the