参附改善大鼠肝移植缺血再灌注后Kupffer细胞激活的研究.docVIP

参附改善大鼠肝移植缺血再灌注后Kupffer细胞激活的研究 柴跃龙,龚建平,涂 兵 （400010重庆，重庆医科大学附属第二医院肝胆外科） [摘要] 目的 观察参附（Shenfu,SF）注射液预处理对大鼠肝移植缺血再灌注后Kupffer细胞中糖皮质激素受体 glucocorticoid receptor,B（NF-κB）表达的影响，探讨参附预处理保护缺血再灌注损害（ischemia reperfusion injury，IRI）的机制。 方法 将雄性SD大鼠按完全随机分组法分为正常对照组、缺血再灌注组、SF组，其中缺血再灌注组、SF组建立肝移植缺血再灌注模型，于缺血再灌注后6h分离培养受体移植肝脏的Kupffer细胞。通过RT-PCR、Western blot和激光共聚焦显微镜技术、免疫荧光法、ELISA等实验技术检测Kupffer细胞GR的mRNA和蛋白表达，NF-κB的蛋白表达，培养上清中TNF-α的含量。结果 缺血再灌注组Kupffer细胞GR mRNA和蛋白表达均明显低于正常对照组（P 0.01），缺血再灌注组NF-κB、培养上清TNF-α表达量 830.19±71.34 明显高于正常对照组NF-κB、培养上清TNF-α表达量（65.41±37.63，P 0.01）。与缺血再灌注组相比，SF组GR水平明显上升（P 0.01），NF-κB活性、TNF-α表达量 543.59±41.27 明显下降（P 0.01）。结论 缺血再灌注损伤会导致Kupffer细胞GR的表达下调，NF-κB的激活；参附预处理可以改善肝移植缺血再灌注后Kupffer细胞的激活。 [关键词] Kupffer细胞；肝移植；缺血再灌注损伤；参附；糖皮质激素受体；核因子-κB [中图法分类号] [文献标志码] A Regulation of Shenfu on activation of Kupffer cells on ischemia-reperfusion injury of transplanted livers in vitro experiments of rates Chai Yuelong, Gong Jianping, Tu Bing Department of Hepatobiliary Surgery, Second Affiliated Hospital, Chongqing Medical University,Chongqing,400010, China [Abstract] Objective To observe the protective effects of Shenfu injection SF pretreatment on the expressions of glucocorticoid receptor GR and nuclear factor kappa B NF-κB in Kupfer cells KCs after liver transplantation in order to elucidate the protective mechanism of SF pretreatment against ischemia-reperfusion injury IRI . Methods SD rats were randomly divided into there groups: the control group n 6 did not perform any surgery, donor animals of the the SF group n 6 were treated with SF（2ml/100g, iv）1h before surgery, and the ischemia-reperfusion group IR n 6 were treated with physiological saline. KCs were isolated from normal rat liver and liver grafts at 6h after transplantation. The expression of GR, NF-κB in KCs and TNF-α secretion were measured by RT-PCR, Western blot ,laser confocal scanning microscope and ELISA. Results The mRNA and protein expression of GR in the IR group were significantly lower P 0.