中国危重病急救医学
中國危重病急救醫學
중국위중병급구의학
CHINESE CRITICAL CARE MEDICINE
2011年
2期
72-76
,共5页
内皮祖细胞%脓毒症%炎症反应%组织损伤
內皮祖細胞%膿毒癥%炎癥反應%組織損傷
내피조세포%농독증%염증반응%조직손상
Endothelial progenitor cell%Sepsis%Inflammatory reaction%Tissue damage
目的 观察并分析脓毒症时循环内皮祖细胞(cEPCs)数量变化及其意义.方法 采用盲肠结扎穿孔术制备雄性SD大鼠脓毒症模型(80只),设正常对照组(16只)、假手术组(80只).于制模后即刻、6、12、18 h及1、2、3、7 d各组取9只大鼠,动态观察外周血单个核细胞(PBMCs)中cEPCs数(流式细胞学法)及血中肿瘤坏死因子-α(TNF-α)、白细胞介素-10(IL-10)、D-二聚体[酶联免疫吸附法(ELISA)]和抗凝血酶-Ⅲ(AT-Ⅲ,免疫浊度法)水平,肝、肾、肺组织湿/干重(W/D)比值.制模后1 d各组取8只大鼠,观察肝、肾、肺组织病理学变化及损伤评分.结果 制模后脓毒症大鼠cEPCs数量明显升高,于18 h达高峰[(7 161.9±689.8)个/106 PBMCs];血中TNF-α、IL-10、D-二聚体、AT-Ⅲ升高,分别于制模后12 h、12 h、3 d、18 h达峰值[(51.3±6.8) ng/L、(77.9±8.6) ng/L、(93.5±11.5) mg/L、(193.8±43.0) mg/L];肝、肾、肺W/D比值和组织损伤评分增加[18 h W/D比值:肝3.79±0.09,肾4.25±0.08,肺4.91±0.09;1 d组织损伤评分(分):肝1.86±0.26,肾5.14±0.34,肺6.57±0.37].模型组上述各指标均显著高于同期假手术组[18 h cEPCs数量(2 235.5±472.7)个/106 PBMCs,12 h TNF-α (14.3±5.8) ng/L,12 h IL-10 (35.0±5.8) ng/L,3 d D-二聚体(14.2±4.4) mg/L,18 h AT-Ⅲ (100.1±12.8) mg/L;18 h W/D比值:肝3.50±0.07,肾3.96±0.04,肺4.54±0.14;1 d组织损伤评分(分):肝0.29±0.18,肾0.57±0.20,肺1.14±0.51,P<0.05或P<0.01].相关分析显示,cEPCs数量与TNF-α(r=0.587)、IL-10(r=0.497)、D-二聚体(r=0.294)、AT-Ⅲ(r=0.690)及肝、肾、肺W/D比值(r1=0.532、r2=0.532、r3=0.679)均呈正相关(均P＜0.01).结论 脓毒症时cEPCs数量明显升高,其变化与炎症反应、凝血激活、毛细血管渗漏和组织损伤程度呈正相关.提示cEPCs数量增加是机体对脓毒症的反应,且数量变化可能代表炎症反应和内皮与组织损伤的程度.
目的 觀察併分析膿毒癥時循環內皮祖細胞(cEPCs)數量變化及其意義.方法 採用盲腸結扎穿孔術製備雄性SD大鼠膿毒癥模型(80隻),設正常對照組(16隻)、假手術組(80隻).于製模後即刻、6、12、18 h及1、2、3、7 d各組取9隻大鼠,動態觀察外週血單箇覈細胞(PBMCs)中cEPCs數(流式細胞學法)及血中腫瘤壞死因子-α(TNF-α)、白細胞介素-10(IL-10)、D-二聚體[酶聯免疫吸附法(ELISA)]和抗凝血酶-Ⅲ(AT-Ⅲ,免疫濁度法)水平,肝、腎、肺組織濕/榦重(W/D)比值.製模後1 d各組取8隻大鼠,觀察肝、腎、肺組織病理學變化及損傷評分.結果 製模後膿毒癥大鼠cEPCs數量明顯升高,于18 h達高峰[(7 161.9±689.8)箇/106 PBMCs];血中TNF-α、IL-10、D-二聚體、AT-Ⅲ升高,分彆于製模後12 h、12 h、3 d、18 h達峰值[(51.3±6.8) ng/L、(77.9±8.6) ng/L、(93.5±11.5) mg/L、(193.8±43.0) mg/L];肝、腎、肺W/D比值和組織損傷評分增加[18 h W/D比值:肝3.79±0.09,腎4.25±0.08,肺4.91±0.09;1 d組織損傷評分(分):肝1.86±0.26,腎5.14±0.34,肺6.57±0.37].模型組上述各指標均顯著高于同期假手術組[18 h cEPCs數量(2 235.5±472.7)箇/106 PBMCs,12 h TNF-α (14.3±5.8) ng/L,12 h IL-10 (35.0±5.8) ng/L,3 d D-二聚體(14.2±4.4) mg/L,18 h AT-Ⅲ (100.1±12.8) mg/L;18 h W/D比值:肝3.50±0.07,腎3.96±0.04,肺4.54±0.14;1 d組織損傷評分(分):肝0.29±0.18,腎0.57±0.20,肺1.14±0.51,P<0.05或P<0.01].相關分析顯示,cEPCs數量與TNF-α(r=0.587)、IL-10(r=0.497)、D-二聚體(r=0.294)、AT-Ⅲ(r=0.690)及肝、腎、肺W/D比值(r1=0.532、r2=0.532、r3=0.679)均呈正相關(均P＜0.01).結論 膿毒癥時cEPCs數量明顯升高,其變化與炎癥反應、凝血激活、毛細血管滲漏和組織損傷程度呈正相關.提示cEPCs數量增加是機體對膿毒癥的反應,且數量變化可能代錶炎癥反應和內皮與組織損傷的程度.
목적 관찰병분석농독증시순배내피조세포(cEPCs)수량변화급기의의.방법 채용맹장결찰천공술제비웅성SD대서농독증모형(80지),설정상대조조(16지)、가수술조(80지).우제모후즉각、6、12、18 h급1、2、3、7 d각조취9지대서,동태관찰외주혈단개핵세포(PBMCs)중cEPCs수(류식세포학법)급혈중종류배사인자-α(TNF-α)、백세포개소-10(IL-10)、D-이취체[매련면역흡부법(ELISA)]화항응혈매-Ⅲ(AT-Ⅲ,면역탁도법)수평,간、신、폐조직습/간중(W/D)비치.제모후1 d각조취8지대서,관찰간、신、폐조직병이학변화급손상평분.결과 제모후농독증대서cEPCs수량명현승고,우18 h체고봉[(7 161.9±689.8)개/106 PBMCs];혈중TNF-α、IL-10、D-이취체、AT-Ⅲ승고,분별우제모후12 h、12 h、3 d、18 h체봉치[(51.3±6.8) ng/L、(77.9±8.6) ng/L、(93.5±11.5) mg/L、(193.8±43.0) mg/L];간、신、폐W/D비치화조직손상평분증가[18 h W/D비치:간3.79±0.09,신4.25±0.08,폐4.91±0.09;1 d조직손상평분(분):간1.86±0.26,신5.14±0.34,폐6.57±0.37].모형조상술각지표균현저고우동기가수술조[18 h cEPCs수량(2 235.5±472.7)개/106 PBMCs,12 h TNF-α (14.3±5.8) ng/L,12 h IL-10 (35.0±5.8) ng/L,3 d D-이취체(14.2±4.4) mg/L,18 h AT-Ⅲ (100.1±12.8) mg/L;18 h W/D비치:간3.50±0.07,신3.96±0.04,폐4.54±0.14;1 d조직손상평분(분):간0.29±0.18,신0.57±0.20,폐1.14±0.51,P<0.05혹P<0.01].상관분석현시,cEPCs수량여TNF-α(r=0.587)、IL-10(r=0.497)、D-이취체(r=0.294)、AT-Ⅲ(r=0.690)급간、신、폐W/D비치(r1=0.532、r2=0.532、r3=0.679)균정정상관(균P＜0.01).결론 농독증시cEPCs수량명현승고,기변화여염증반응、응혈격활、모세혈관삼루화조직손상정도정정상관.제시cEPCs수량증가시궤체대농독증적반응,차수량변화가능대표염증반응화내피여조직손상적정도.
Objective To observe the change in number of circulating endothelial progenitor cells (cEPCs) and analyze its significance in septic rat. Methods Septic model of male Sprague-Dawley (SD) rats was reproduced by cecum ligation and puncture (n=80), and the normal control group (n=16) and sham operation group (n=80) were established. Nine rats in each group were used, and the cEPCs numbers in peripheral blood mononuclear cells (PBMCs, by flow cytometry), tumor necrosis factor-α (TNF-α), interleukin-10 (IL-10), D-dimer (by enzyme linked immunosorbent assay, ELISA), antithrombase-Ⅲ (AT-Ⅲ, by immunonephelometry), wet/dry (W/D) ratio of liver, kidney and lung were determined at 0, 6, 12, 18 hours and 1, 2, 3, 7 days after reproduction of model. Eight rats in each group were used, and the pathologic changes in liver, kidney and lung at 1 day were observed, and the injury scores were evaluated. Results The cEPCs number was markedly increased, reaching the peak [(7 161.9±689.8)/106 PBMCs] at 18 hours. Circulating TNF-α, IL-10, D-dimer, AT-Ⅲ were found to be increased, and the levels reached the peak at 12 hours, 12 hours, 3 days, 18 hours, respectively [(51.3±6.8) ng/L, (77.9±8.6) ng/L, (93.5±11.5) mg/L, (193.8±43.0) mg/L]. W/D ratio was elevated and signs of injury to the liver, kidney, lung became more obvious (18-hour W/D of liver: 3.79±0.09, kidney: 4.25±0.08, lung: 4.91±0.09; 1-day tissue evaluation of liver: 1.86±0.26, kidney: 5.14±0.34, lung: 6.57±0.37). The levels of all parameters in model group were significantly higher than those in sham operation group [18-hour cEPCs numbers: (2 235.5±472.7)/106 PBMCs, 12-hour TNF-α: (14.3±5.8) ng/L, 12-hour IL-10: (35.0±5.8) ng/L, 3-day D-dimer: (14.2±4.4) mg/L, 18-hour AT-Ⅲ: (100.1±12.8) mg/L; 18-hour liver W/D ratio: 3.50±0.07, kidney: 3.96±0.04, lung: 4.54±0.14; 1-day tissue evaluation of liver: 0.29±0.18, kidney: 0.57±0.20, lung: 1.14±0.51, P<0.05 or P<0.01]. There was positive correlation between cEPCs numbers and TNF-α (r=0.587), IL-10 (r=0.497), D-dimer (r=0.294), AT-Ⅲ (r=0.690), and W/D ratio of liver, kidney, lung (r1=0.532, r2=0.532, r3=0.679, all P<0.01). Conclusion The cEPCs number markedly increases in septic rats, and it shows positive correlation with the degree of inflammatory reaction, blood clotting activation, capillary leakage and tissue damage. The increase of number of cEPCs is the result of reaction to sepsis, and its change in number might be valuable in evaluating the pathogenesis of sepsis.
