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1、尼莫地平抑制大鼠燒傷后枯否細(xì)胞TNFα的產(chǎn)生【關(guān)鍵詞】燒傷尼莫地平枯否細(xì)胞腫瘤壞死因子【摘要】目的探討鈣離子通道阻斷劑尼莫地平調(diào)控?zé)齻罂莘窦?xì)胞(KC)合成釋放TNFα的可能性,為尋找到一種有效減輕、控制燒傷后過度全身炎癥反應(yīng)的措施提供理論依據(jù)。方法內(nèi)灌注消化、密度梯度離心法分離培養(yǎng)正常SD大鼠KC,顯微熒光分光光度計復(fù)合倒置顯微鏡技術(shù)觀察燙傷血清作用下單個KC細(xì)胞內(nèi)鈣([Ca2+]i)變化,ELISA方法測定燙傷血清培養(yǎng)的KC上清中TNFα濃度變化;SD大鼠行30%TBSAⅢ度燙傷,
2、傷后6h分離KC,RNA酶保護(hù)分析法測定KCTNFαmRNA表達(dá)量,并測定血漿TNFα水平;觀察尼莫地平存在時,上述結(jié)果的改變。結(jié)果與對照組相比,燒傷組KC[Ca2+]i峰值及培養(yǎng)上清中TNFα濃度增加值均顯著增加(P<0.01),在1μM尼莫地平存在時,兩者均顯著減少(P<0.01)。燒傷后6hKCTNFαmRNA表達(dá)量及血漿TNFα水平顯著升高,靜脈給予尼莫地平(40μg?kg-1?h-1)后,兩者均顯著減少(P<0.01)。結(jié)論燒傷后,KC合成釋放TNFα,通過細(xì)胞內(nèi)鈣離子通道信號傳導(dǎo)途徑實(shí)現(xiàn)。尼莫地平能抑制燒傷后KCTNFαmRN
3、A表達(dá),使KC產(chǎn)生TNFα明顯減少,并下調(diào)血漿中TNFα的總體水平。關(guān)鍵詞燒傷尼莫地平枯否細(xì)胞腫瘤壞死因子αNimodipinecaninhibittheproductionofTNFαbykupffercellsinsevereburnedratsWangGuangyi,TianJianguang,ZhuShihui,etal.
4、">BurnDepartment,ChanghaiHospital,Shanghai200433.【Abstract】ObjectiveTostudywhethernimodipine,aDihydropyridine-typecalciumchannelblocker,caninhibittheproductionofTNFαbykupffercells(KC)anddown-regulateitslevelofplasmaaftersevereburninjury.MethodsK
5、Csofnormalratswereisolatedwithportalveincatheter,intrahepaticdigestionanddensitygradientcenˉtrifugation.Intracellularcalciumconcentration([Ca2+]i)inindividualKCafterstimulatedwithpostburnserumwasasˉsessedfluorometricallywithmicrospectrofluorometer.LevelofTNFαinthesupernat
6、antofKCculturedwithpostburnserumwasdetectedbyELISA.SDratsunderwent30%TBSAfullthicknessburn.Sixhourslater,KCwereisolatedandtheirtotalRNAwasextracted.LevelofTNFαmRNAwasdetectedbyribonucleaseprotectionassay.LevelofplasmaTNFαwasalsodetected.Roleofnimodipineonabove-mentionedef
7、fectswereobserved.ResultsComparedwiththatofcontrolgroup,[Ca2+]iofKCandlevelofTNFαinsupernatantinburngroupincreasedsignificantly.Atpresentof1μMnimodipine,however,thenumericalvaluedecreasedsignificantly.Comparedwiththatofcontrolgroup,levelofKCmRNAandplasmaTNFαinburngroupals
8、oincreasedsignificantly.Afterintravenouslyinjectionwithnimodipine(40μg?kg-1?h-1),thenumericalval