大黃素對模擬冷缺血再灌注后肝細胞內鈣離子濃度及細胞凋亡的影響

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1、大黃素對模擬冷缺血再灌注后肝細胞內鈣離子濃度及細胞凋亡的影響作者：祁翔，呂毅，沈乃營，劉昌，劉學民，王博祁翔【摘要】目的研究中藥大黃素對模擬冷缺血再灌注后肝細胞內鈣離子濃度及細胞凋亡的影響。方法體外培養(yǎng)肝細胞株HL7702，隨機分為對照組和大黃素處理組。對照組未予大黃素處理，大黃素處理組按100、10、1μmol/L分為高、中、低3個濃度組，建立模擬冷缺血再灌注模型，流式細胞技術檢測各組細胞內鈣離子濃度及細胞凋亡水平，分別檢測各組細胞培養(yǎng)上清液乳酸脫氫酶水平。結果冷缺血8h再灌注6h后，高、中、低濃度大黃素處理組鈣離子熒光強度分別為(24.12±0.51)、(26.35±1

2、.34)、(39.12±1.94)，均顯著低于對照組的105.29±1.01(P<0.01)。高、中、低濃度大黃素處理組細胞凋亡率分別為(5.46±0.41)%、(10.64±0.64)%、(11.90±0.50)%，均顯著低于對照組的(25.40±1.41)%(P<0.01)。高、中濃度大黃素處理組上清液LDH含量分別為(179.67±18.57)u/L、(198.83±14.22)u/L，顯著低于對照組的(351.33±34.16)u/L(P<0.01)。結論大黃素可降低模擬冷缺血再灌注后的肝細胞內鈣離子濃度，抑制細胞凋亡，減輕肝細胞損傷。【關鍵詞】大黃

3、素；肝細胞；冷缺血再灌注；細胞內鈣離子濃度；凋亡11　　ABSTRACT:ObjectiveToinvestigatetheeffectofEmodinonintracellularcalciumconcentration([Ca2+]i)andapoptosisofhepaticcellsaftersimulatedcoldischemiareperfusion.MethodsGlucoseoxygendeprivation,lowtemperature,subsequentreoxygenationandrewarmingwereusedtoinduceischemi

4、areperfusioninjurymodelinculturedhepaticcellswhichweredividedinto4groups:controlgroupandEmodintreatedgroup(100,10and1μmol/Lgroups).Theintracellularcalciumconcentration([Ca2+]i)andapoptosisrateweredeterminedbyflowcytometry(FCM)respectively;thecontentoflactatedehydrogenase(LDH)insupernatant

5、wastested.ResultsIntracellularcalciumfluorescenceintensityinEmodintreatedgroupsofhigh,mediumandlowdensitywas24.12±0.51,26.35±1.34and39.12±1.94,respectively,whichweresignificantlylowerthan105.29±1.01incontrolgroup(P<0.01).ApoptosisrateinEmodintreatedgroupsofhigh,mediumandlowdensitywas0.

6、0546±0.0041,0.1064±0.0064and0.1190±0.0050,respectively,whichweresignificantlylowerthan0.2540±0.0141incontrolgroup(P<0.01).LDHwas(198.83±14.22)u/Land(179.67±18.57)u/LinEmodintreatedgroupsofmediumandhighdensityrespectively,whichwere11significantlylowerthan(351.33±34.16)u/Lincontrolgroup(P

7、<0.01).ConclusionEmodincouldreduce[Ca2+]iandinhibitapoptosisofhepaticcellsaftersimulatedcoldischemiareperfusion,thusprotectinghepaticcellseffectively.　　KEYWORDS:Emodin;hepaticcell;ischemiareperfusioninjury;intracellularfreecalciumconcentration;apopt