摘要
目的了解外源性一氧化碳释放分子2（CORM-2）对脓毒症时组织因子（TF）表达的抑制作用。方法培养人脐静脉内皮细胞（HUVEC）并分为正常对照组、LPS组（用10μg／mL LPS孵育，浓度下同）、LPS＋二甲亚砜组及LPS＋10μmol／L CORM-2组、LPS＋50μmol／LCORM-2组、LPS＋100μmol／LCORM-2组，培养4h后检测HUVEC的TF活性、TF蛋白表达和核因子κB的活性。将45只雄性C57 BL／6小鼠随机分成健康对照组5只、盲肠结扎和穿孔术（CLP）组20只和CLP＋CORM-2组20只。CLP＋CORM-2组除伤后注射8．0mg／kg CORM-2外，其他处理与CLP组相同。于术后2、6、12、24h（每时相点5只小鼠）检测血浆TF、TF途径抑制物（TFPI）水平，同时检测健康对照组相应指标。结果与正常对照组比较，LPS组HUVEC的TF活性明显升高（P〈0．05），TF蛋白表达增加，核因子κB活性增强；LPS联合3种不同浓度CORM-2处理组的TF活性呈浓度依赖性下降，核因子κB活性、TF蛋白表达减弱。CLP组小鼠血浆TF水平从术后6h[（80．0±11．9）pg／mL]开始上升，明显高于健康对照组[（58．4±6．9）pg／mL，P〈0．05]；术后24h开始下降，但仍高于健康对照组。CLP组血浆TFPI水平未见明显变化。与健康对照组[（12．4±2．8）ng／mL]比较，CLP＋CORM-2组小鼠术后6、12h血浆TFPI水平[分别为（23．7±3．5）、（24．4±5．0）ng／mL]均明显升高（P〈0．05）。结论外源性CORM-2能明显抑制TF活性，减少TF蛋白表达，抑制核因子κB活性；同时明显减少脓毒症时血浆TF水平，提高TFPI水平，有效防止凝血系统活化，维持促凝、抗凝系统的平衡。
Objective To explore the inhibitive effects of exogenous carbon monoxide-releasing molecules 2（CORM-2） on expression of tissue factor （TF） in sepsis. Methods Human umbilical vein endothelial cells （HUVEC） were cultured with trypsin digestion method, which were divided into NC group （with normal treatment）, LPS group（ with cuhure of 10 μg /mL LPS）, LD group （with 10μg /mL LPS and DMSO in co-culture） , LC1 group（with 10 μg/mL LPS and 10 μmol/L CORM-2 in co-culture） , LC2 group（with 10 μg/mL LPS and 50 μmol/L CORM-2 in co-culture） , LC3 group（with 10 μg/mL LPS and 100 μmol/L CORM-2 in co-culture）. After culture for 4 hours, TF activity, TF protein expression, nuclear factor-κB（NF-κB） activity were examined. Forty-five C57 BL/6 male mice were randomly divided into NC （ without treatment, n = 5 ）, CLP （ n = 5 ） and CLP ＋ CORM-2 （ with treatment of 8 mg/kg CROM-2 after CLP, n =5）groups. The serum samples in CLP, CLP ＋ CORM-2 groups were collected at 2, 6, 12 and 24 post operation hour（ POH, 5 mice at each time point） for determination of TF and TFPI levels,which were also examined in NC group. Results Compared with those of NC group, TF activity increased （ P 〈 0. 01 ） , TF protein expression and NF-κB activity also increased in LPS group. Compared with those of LPS group, above indices were decreased in LC1, LC2, LC3 groups. The serum level of TF in CLP group at 6 POH was higher than that of NC group （80.0 ± 11.9 pg/mL vs 58.4 ±6.9 pg/mL, P 〈0.05）, peaked at 12 POH, and still higher than that of NC group at 24 POH, while the serum level of TFPI showed no obvious difference in NC and CLP groups. Compared with that of NC group, TFPI levels obviously increased in CLP ＋ CORM -2 group at 6, 12 POH（23.7±3.5 ng/mL,24.4 ±5.0 ng/mL vs 12.4±2.8 ng/mL, P 〈0.05）. Conclusions Exogenous CORM can obviously inhibit TF and NF-κB activity,decrease TF protein expres- sion. Meanwhile, it can also decrease serum level of TF, and increase serum level of TFPI, preventing acti-vation of procoagulant system, balancing procoagulant and anticoagulant system in sepsis.
出处
《中华烧伤杂志》
CAS
CSCD
北大核心
2009年第2期111-114,共4页
Chinese Journal of Burns
基金
国家自然科学基金（30772256）
江苏省自然科学基金（BK2008237）
