摘要
目的探讨创伤弧菌脓毒症大鼠脑组织转位蛋白（TSPO）基因和蛋白表达及促／抗炎因子的蛋白表达，并观察抗菌药物头孢哌酮钠联合左氧氟沙星的干预效应。方法SD大鼠100只，随机（随机数字法）分为健康对照组（A组，n=10）、创伤弧菌脓毒症组（B组，n=40）和抗菌药物干预组（C组，n=40），抗菌药物对照组（D组，n=10）。大鼠左后肢皮下注射创伤弧菌（6×10^8cfu／ml，0．1ml／100g）制作脓毒症大鼠模型，腹腔注射头孢派酮钠180ms／kg和左氧氟沙星18ms／kg进行干预，以RT—PCR、Western blotting、免疫组化、ELISA等方法观察大鼠染菌后6、12、24、48h脑组织TSPO表达、促／抗炎因子水平及电镜的特点。结果与A组比较，B组大鼠脑组织TSPO表达水平6h即明显增高（P〈0．05），24h达峰值。C组大鼠12、24、48h脑组织TSPO表达水平均明显低于相同时间点B组大鼠（P〈0．05）。B组大鼠脑组织各时间点TNF-α、IL-6、IL-10水平均比A组明显升高（P〈0．05），其中TNF-α在6h达峰值，IL-6 12h达峰值，IL-10则48h达峰值。与B组相同时间点比较，C组大鼠6、12、24h时点脑组织TNF-α水平明显下降（P〈0．05），12、24、48h时间点脑组织IL-6水平亦明显下调（P〈0．05），而IL-10水平在各时间点均较B组明显升高（P〈0．05）。免疫组化法观察B组大鼠脑组织TSPO蛋白棕黄色颗粒密集较多，C组棕黄色颗粒较B组减少；电镜下B组大鼠脑组织神经元细胞核溶解消失，细胞及细胞器肿胀明显，C组大鼠脑组织神经元细胞核部分溶解，形态尚存，细胞及细胞器水肿减轻。结论创伤弧菌脓毒症大鼠脑组织TSPO的表达随着脑组织炎症、病理进展逐渐增高，能敏感反映脑损伤的变化。使用头孢哌酮钠和左氧氟沙星能有效减轻脑组织炎症损伤，降低TSPO的水平。
Objective To investigate the potential role and changes of translocator protein 18 kDa （TSPO） in brain tissue of vibrio vulnificus （VV） septic rats and effects of Cefoperazone Sodium combined with Levofloxaein. Methods One hundred SD rats were divided randomly （random number） into four groups ： normal control group （ A, n = 10 ）, VV sepsis model group （ B, n = 40 ）, antibiotics treatment group （ C, n = 40）, and antibiotics control group （ D, n = 10 ）. Septic model was replicated by injection with VV （6 × 108 cfu/ml, 0. 1 ml/100 g） in the left posterior leg of rats. The antibacterials were given intraperitoneally with Cefoperazone sodium （180 mg/ kg） combined with Levofloxacin （18 mg/kg）. The level of TSPO and pro-and anti-Inflammatory cytokines in rattish brain at 6, 12, 24, 48 h after infection were observed by RT-PCR, Western blotting, immunohistochemisty and ELISA. Then pathological change in brain tissue was detected by electron microscope. Results Compared with group A, the expression of TSPO in brain tissue was significantly increased in group B （P 〈 0. 05 ） , and reached the peak at 24 h. The TSPO expression in group C was significantly decreased compared with that in group B at 12, 24, 48 h （P 〈 0. 05）. The levels of TNF-α, IL-6, IL-10 in brain tissue of group B were significantly higher than those in group A （P 〈 0. 05 ）. Compared with group B, the TNF-α expression in group C was siginificantly decreased at 6, 12, 24 h （P 〈 0. 05 ） , while the IL-6 expression was siginificantly decreased at 12, 24, 48 h （P 〈 0. 05） and IL-10 level was significantly increased at 6, 12, 24, 48 h （P 〈 0. 05）. Immunohistochemical tests showed brownish yellow particles （TSPO protein） in the brain tissue of group B were more dense than those in group C. Electron microscope showed the nerve cells and organelles were swelling intensively and the neuronal nucleus were dissolved and disappear in group B. But the neuronal nucleus were dissolved partly and the swelling were alleviative in group C. Conclusions TSPO expression is markedly increased in accord with brain tissue injury and inflammation, which makes it suited for assessing brain injury as a sensitive biomarker in VV sepsis. Effective antibiotics can inhibit the inflammatory and injury and reduce the level of TSPO in septic brain.
出处
《中华急诊医学杂志》
CAS
CSCD
北大核心
2013年第3期260-265,共6页
Chinese Journal of Emergency Medicine
基金
浙江省医学创新学科建设计划（11-CX26）
浙江省十二五高校重点学科建设计划
温州市科技计划项目（Y20090111）
关键词
创伤弧菌
脓毒症脑病
转位蛋白
抗菌药物
大鼠
Vibro vulnificus
Sepsis encephalopathy
Translocator protein （TSPO）
Antibacterials
Rat
