中国危重病急救医学
中國危重病急救醫學
중국위중병급구의학
CHINESE CRITICAL CARE MEDICINE
2012年
8期
487-489
,共3页
郑贵军%武子霞%朱海云%殷冬梅%孙茜%李银平
鄭貴軍%武子霞%硃海雲%慇鼕梅%孫茜%李銀平
정귀군%무자하%주해운%은동매%손천%리은평
脂多糖%主动脉内皮细胞%活化蛋白C%血管性血友病因子%血管性血友病因子裂解酶%大鼠
脂多糖%主動脈內皮細胞%活化蛋白C%血管性血友病因子%血管性血友病因子裂解酶%大鼠
지다당%주동맥내피세포%활화단백C%혈관성혈우병인자%혈관성혈우병인자렬해매%대서
Lipopolysaccharide%Aortic endothelial cell%Activated protein C%Von Willebrand factor%Von Willebrand factor cleaving protease%Rat
目的 探讨活化蛋白C(APC)对脂多糖(LPS)诱导大鼠主动脉内皮细胞(RAECs)血管性血友病因子抗原( vWFAg)及其裂解酶(ADAMTS-13)蛋白表达的影响.方法 采用组织贴块法培养Wista护大鼠的RAECs,1周后传代至第4～5代用于实验.将细胞分为对照组、LPS刺激组(1 mg/L)以及APC干预组(在LPS刺激后分别加入终浓度为0.1、1、10 mg/L的APC)；分别于12、24、48、72 h采用酶联免疫吸附试验(ELISA)测定RAECs培养上清液中vWFAg和ADAMTS-13蛋白的表达水平.结果 对照组仅有少量的vWFAg和ADAMTS-13蛋白表达.随LPS刺激时间延长,vWFAg表达12h即明显增多,48 h达峰值[(285.45±30.13)％],ADAMTS-13蛋白表达(μg/L)呈降低趋势,72 h降至最低(13.32±2.37),与对照组[vWFAg:(94.53±7.83)％,ADAMTS-13:115.76±2.36]比较差异有统计学意义(均P＜0.01).APC可逆转LPS刺激后对vWFAg的促进作用和对ADAMTS- 13的抑制作用,且剂量越大,逆转效果越明显.10 mg/LAPC干预后可使LPS刺激48 h时达峰值的vWFAg明显降低[(198.43±17.92)％比(285.45±30.13)％],使LPS刺激72 h时降至最低的ADAMTS-13(μg/L)明显升高(125.25±2.70比13.32±2.37),差异有统计学意义(均P＜0.01).结论 LPS刺激RAECs后vWFAg明显升高,ADAMTS-13蛋白表达明显降低,具有时间依赖性；用不同浓度APC干预后,随浓度增加和作用时间延长,vWFAg明显降低,ADAMTS-13蛋白表达明显升高,具有剂量依赖性和时间依赖性.
目的 探討活化蛋白C(APC)對脂多糖(LPS)誘導大鼠主動脈內皮細胞(RAECs)血管性血友病因子抗原( vWFAg)及其裂解酶(ADAMTS-13)蛋白錶達的影響.方法 採用組織貼塊法培養Wista護大鼠的RAECs,1週後傳代至第4～5代用于實驗.將細胞分為對照組、LPS刺激組(1 mg/L)以及APC榦預組(在LPS刺激後分彆加入終濃度為0.1、1、10 mg/L的APC)；分彆于12、24、48、72 h採用酶聯免疫吸附試驗(ELISA)測定RAECs培養上清液中vWFAg和ADAMTS-13蛋白的錶達水平.結果 對照組僅有少量的vWFAg和ADAMTS-13蛋白錶達.隨LPS刺激時間延長,vWFAg錶達12h即明顯增多,48 h達峰值[(285.45±30.13)％],ADAMTS-13蛋白錶達(μg/L)呈降低趨勢,72 h降至最低(13.32±2.37),與對照組[vWFAg:(94.53±7.83)％,ADAMTS-13:115.76±2.36]比較差異有統計學意義(均P＜0.01).APC可逆轉LPS刺激後對vWFAg的促進作用和對ADAMTS- 13的抑製作用,且劑量越大,逆轉效果越明顯.10 mg/LAPC榦預後可使LPS刺激48 h時達峰值的vWFAg明顯降低[(198.43±17.92)％比(285.45±30.13)％],使LPS刺激72 h時降至最低的ADAMTS-13(μg/L)明顯升高(125.25±2.70比13.32±2.37),差異有統計學意義(均P＜0.01).結論 LPS刺激RAECs後vWFAg明顯升高,ADAMTS-13蛋白錶達明顯降低,具有時間依賴性；用不同濃度APC榦預後,隨濃度增加和作用時間延長,vWFAg明顯降低,ADAMTS-13蛋白錶達明顯升高,具有劑量依賴性和時間依賴性.
목적 탐토활화단백C(APC)대지다당(LPS)유도대서주동맥내피세포(RAECs)혈관성혈우병인자항원( vWFAg)급기렬해매(ADAMTS-13)단백표체적영향.방법 채용조직첩괴법배양Wista호대서적RAECs,1주후전대지제4～5대용우실험.장세포분위대조조、LPS자격조(1 mg/L)이급APC간예조(재LPS자격후분별가입종농도위0.1、1、10 mg/L적APC)；분별우12、24、48、72 h채용매련면역흡부시험(ELISA)측정RAECs배양상청액중vWFAg화ADAMTS-13단백적표체수평.결과 대조조부유소량적vWFAg화ADAMTS-13단백표체.수LPS자격시간연장,vWFAg표체12h즉명현증다,48 h체봉치[(285.45±30.13)％],ADAMTS-13단백표체(μg/L)정강저추세,72 h강지최저(13.32±2.37),여대조조[vWFAg:(94.53±7.83)％,ADAMTS-13:115.76±2.36]비교차이유통계학의의(균P＜0.01).APC가역전LPS자격후대vWFAg적촉진작용화대ADAMTS- 13적억제작용,차제량월대,역전효과월명현.10 mg/LAPC간예후가사LPS자격48 h시체봉치적vWFAg명현강저[(198.43±17.92)％비(285.45±30.13)％],사LPS자격72 h시강지최저적ADAMTS-13(μg/L)명현승고(125.25±2.70비13.32±2.37),차이유통계학의의(균P＜0.01).결론 LPS자격RAECs후vWFAg명현승고,ADAMTS-13단백표체명현강저,구유시간의뢰성；용불동농도APC간예후,수농도증가화작용시간연장,vWFAg명현강저,ADAMTS-13단백표체명현승고,구유제량의뢰성화시간의뢰성.
Objective To investigate the activated protein C (APC) on the von Willebrand factor antigen (vWFAg) and von Willebrand factor cleaving protease ( ADAMTS-13 ) protein expression in rat aortic endothelial cells (RAECs) induced by lipopolysaccharide (LPS).Methods RAECs from Wistar rats were cultured with the tissue explants adherence method.RAECs were cultured for one week,After one week culture,RAECs in 4-5 generations were divided into control group,LPS stimulation groups ( 1mg/L) and APC intervention groups (0.1,1 and 10 mg/L APC was added after LPS stimulation).The supernatants were obtained at 12,24,48,and 72 hours after LPS stimulated to determine the vWFAg and protein of ADAMTS-13 expression by enzyme-linked immunoadsorbent assay (ELISA).Results In the control group,RAECs expressed little vWFAg and protein of ADAMTS-13.With stimulation of LPS,the vWFAg was significantly increased at 12 hours,and reached the peak at 48 hours [(285.45 ± 30.13 )％],and the level of ADAMTS-13 (μg,/L) was gradually decreased,and reached the nadir at 72 hours ( 13.32 ± 2.37 ),there was significant difference compared with control group [vWFAg: (94.53 ± 7.83 )％,ADAMTS-13:115.76 ±2.36,both P＜0.01 ).The effects on vWFAg promoting and ADAMTS-13 inhibition after LPS stimulation coull be dose-dependently reversed by APC.10 mg/L of APC could decrease the peak of vWFAg at 48 hours of LPS stimulation [ ( 198.43 ± 17.92 )％ vs.( 285.45 ± 30.13 )％ ],and increase the minimize of ADAMTS-13 ( μg/L ) at 72 hours of LPS stimulation (125.25 ±2.70 vs.13.32 ± 2.37),with significant difference (both P＜0.01 ).Conclusions After stimulation with LPS,the level of vWFAg was time-dependent increased,as the protein of ADAMTS-13 was decreased.APC could attenuate the effect of LPS on vWFAg and protein of ADAMTS-13 with dose-dependent and time-dependent patterns.
