摘要
目的观察烧伤后脓毒症患者高迁移率族蛋白-1（HMGB-1）的变化，探讨卡巴胆碱对脂多糖刺激后单核细胞释放HMGB-1的影响及其作用机制。方法取烧伤后脓毒症患者及正常献血员外周血，ELISA法检测HMGB-1含量。观察烧伤后脓毒症对HMGB-1的影响。取正常献血员外周血，分离获得单核细胞。实验分为6组：空白对照组的单核细胞仅加1640培养液；脂多糖组单核细胞仅加脂多糖刺激；烟碱预处理组和卡巴胆碱预处理组先用卡巴胆碱或烟碱预处理单核细胞5min，再加入脂多糖刺激；阿托品＋卡巴胆碱预处理组和α-银环蛇毒素＋卡巴胆碱预处理组先在单核细胞悬液中加入阿托品或α-银环蛇毒素，5min后给予卡巴胆碱，再5min后给予脂多糖刺激。孵育48h后取细胞培养上清液，ELISA法检测HMGB-1浓度。结果烧伤后脓毒症患者HMGB-1的含量[（12．94±6．54）μg/L]明显增加，与正常献血员[（2．01±0．03）μg／L]比较差异有统计学意义（P〈0．01）。脂多糖单独刺激单核细胞时，HMGB-1含量[（9．39±1．37）μg/L]较对照组[（1．48±0．69）μg／L]明显升高（P〈0．01）。用卡巴胆碱或烟碱预处理细胞后，HMGB-1含量明显下降[（3．52±1．64）μg/L与（4．01±1．56）μg／L]，与脂多糖单独刺激组比较差异有统计学意义（P〈0．01）。阿托品预处理细胞后，卡巴胆碱对脂多糖刺激下单核细胞释放HMGB-1的抑制作用无明显变化[（3．87±2．01）μg／L]，而α-银环蛇毒素预处理细胞后，卡巴胆碱对HMGB-1的抑制作用明显减弱[（8．97±1．97）μg／L]。结论烧伤后脓毒症患者HMGB-1释放明显增加，卡巴胆碱能够减少单核细胞释放HMGB-1，其作用机制可能是通过N样胆碱能受体α-7亚基实现的。
Objective To observe the changes of HMGB-1 of burned patient with sepsis and to investigate the effect of carhachol on production of HMGB-1 from human monocytes stimulated by lipopolysaccharide（LPS） and its mechanism. Methods The peripheral blood samples of burned patients with sepsis and healthy donors were taken for isolation of monocyte. The subjects were divided into six groups ： the menocytes group were added only with 1640 culture medium, LPS group were added only with LPS;nicotine group and carbachol group were with carhachol first or nicotine for 5 rain and then with LPS. The levels of HMGB-1 were tested by ELISA. α-bungarotoxin ＋ carbachol group were added with atropine or α-bungarotoxin for 5 min and then were given carbachol and 5 rain later were stimulated with LPS. 48 h-incubation later supernate were collected for the detection of HMGB-1 by ELISA. Results HMGB-1 levels of burned patients with sepsis were （12.94 ±6.54）pfL,which were much higher than that of healthy donors（ [ （2.01 ±0.03） μ/L） ,P 〈0.01 ]. When monocytes were stimulated by LPS alone,concentrations of HMGB-1 were （9.39 ± 1.37） μ/L, which were obviously higher than that of controls [ （ 1.48 ± 0.69） p/L）, P 〈 0.01 ]. After pretreated by carbachol or nicotine, concentrations of HMGB-1 were （3.52 ±1.64） μ/L and （4.01±1.56）μ/L respectively, which were obviously decreased compared with that of LPS stimulation alone group （P 〈 0.01 ）. When pretreatedby atropine before addition of carbachol, there was no significant changes in concentrations of HMGB-1 [ （3.87± 2.01 ） μ/L ]. When pretreated with α-Bungarotoxin before carbachol adminitration, inhibitive effect of carbachol on production of HMGB-I were blocked [ （8.97 ± 1.97） μ/L ]. Conclusion The release of HMGB-1 in burned patients with sepsis is increased. Carbachol could obviously reduce the production of HMGB-1, which may affect through activating α7 subunit of cholinergic N receptor.
出处
《中国综合临床》
2009年第5期544-546,共3页
Clinical Medicine of China
