磷脂酰肌醇特异性磷脂酶C对内毒素血症大鼠肝组织CD14表达的抑制作用  

Inhibitory role of PI-PLC on CD14 expression in rat liver tissue during endotoxemia

作　　者：孙航[1] 吴传新[2] 龚建平[2] 刘海忠[2] 李旭宏[2] 游海波[2] 陈先锋[2] 

机构地区：[1]重庆医科大学病毒性肝炎研究所,四川重庆400010 [2]重庆医科大学第二附属医院肝胆外科,四川重庆400010

出　　处：《基础医学与临床》2005年第4期337-341,共5页Basic and Clinical Medicine

基　　金：国家自然科学基金(3017091930471696)

摘　　要：目的观察磷脂酰肌醇特异性磷脂酶C(PI-PLC)对大鼠内毒素血症时肝组织中Kuffer细胞(KCs)的活性、CD14mRNA的表达和血浆细胞因子释放的作用。方法将Wistar大鼠90只,随机分为LPS组(静注LPS5mg/kg)、PI-PLC组(注LPS前30min注PI-PLC100U/kg)和生理盐水NS组。分别于注射前及注射后1、3、6和12h取材,每组每时相点6只,测定血浆内毒素(鲎试剂基质偶氮显色法)、LBP及TNF-α(均用ELISA法)和IL-6(放免法)的含量,用RT-PCR检测肝组织中CD14mRNA的表达,并观察其形态学变化。结果LPS组LBP、TNF-αI、L-6的含量和CD14mRNA的表达明显增加,并伴有KCs激活,数量增多,体积增大,吞噬功能增强,肝细胞出现变性和坏死等;而PI-PLC处理组所致的上述变化明显减轻。结论PI-PLC对内毒素所致肝组织内KCs的激活有一定抑制作用。Objective To study the inhibitory role of phosphatidylinositol- specific phospholipase C(PI-PLC) on the activation of KCs and CD14 expression in rat liver tissue and plasma cytokine during endotoxemia. Methods Ninety Wistar rats were randomly divided into three groups, LPS group whose rats were injected LPS (5 mg/kg),PI-PLC group whose rats were injected PI-PLC (100 U/kg) (30 min) before injected LPS and NS group whose rats were injected NS (0.2 mL). The rats were sacrificed before and 1, 3, 6 and (12 h) after injection respectively. There were six rats at each time point. Liver tissue was collected to determine CD14 mRNA expression by RT-PCR. The levels of plasma endotoxin, lopopolysaccharide binding protein(LBP), TNF-α and IL-6 were also determined. The pathological changes of liver tissue were observed with optical and electron microscopy. Results When the level of plasma LPS elevated, expression of CD14 mRNA in the liver tissue were stronger and the level of plasma LBP, TNF-α and IL-6 were higher in the PI-PLC group and LPS group than in the NS group(P<0.01), but the expression of CD14 mRNA in the liver tissue and the level of plasma LBP and TNF-α and IL-6 were lower in the PI-PLC group than in the LPS group(P<0.01). KCs were activated in the PI-PLC group and LPS group. These cells proliferated and enlarged. Meanwhile, their surface projection was increased in number and their cytoplasm was full of phagocytic vacuoles or electron dense phagosomes, which indicates active phagocytosis. Focal cytoplasmic degeneration and necrosis could be seen in hepatocytes. But those changes were lighter in the PI-PLC group than in the LPS group. WT10.HZConclusions PI-PLC could inhibit the activation of KCs and the expression of CD14 mRNA in the liver tissue during endotoxemia. And it also could inhibit KCs to produce and release various cytokines during endotoxemia.

关 键 词：内毒素血症 磷脂酰肌醇 抑制作用 肝组织 特异性 磷脂酶C Wistar大鼠 RT-PCR检测 CD14mRNA PI-PLC TNF-α 细胞因子释放 ELISA法 偶氮显色法 血浆内毒素 形态学变化 LPS 生理盐水 IL-6 吞噬功能 LBP KCs 注射后 鲎试剂 

分 类 号：R363[医药卫生—病理学]