联合应用甘氨酸和甲强龙对失血性休克大鼠肝脏枯否细胞的影响

The effect of combination of glycine and methylprednisolone on Kupffer cells of liver after hemorrhagic shock in rats

目的探讨失血性休克大鼠肝脏枯否细胞的功能变化以及联合应用甘氨酸和甲强龙对枯否细胞的影响。方法50只大鼠随机分成假休克组(仅进行手术操作但不放血诱导休克)、休克组、甘氨酸组、甲强龙组和联合治疗组(甘氨酸+甲强龙),每组10只。大鼠经动脉放血,造成失血性休克,随后用自体血和生理盐水回输进行复苏。复苏后2 h,行肝脏枯否细胞的分离和培养,细胞培养24 h后分别用1、10、100和1000 ng/m l的脂多糖(LPS)刺激,测定细胞内Ca2+和肿瘤坏死因子α(TNF-α)水平。结果枯否细胞内Ca2+在20 m in左右开始大幅度增高,大约27 m in达到高峰,同时细胞内Ca2+浓度增加呈现LPS剂量依赖性。联合治疗组细胞内Ca2+浓度和TNF-α含量明显低于休克组以及低于甘氨酸、甲强龙治疗组,差异具有统计学意义(P<0.005)。结论联合应用甘氨酸和甲强龙比单一制剂更有效抑制失血性休克后枯否细胞内Ca2+升高、抑制枯否细胞的激活,抑制TNF-α的过度产生和机体炎症反应。

Objective To investigate the effect of combination of glycine and methylprednisolone （MP） on Kupffer cells in hver of rats suffered from hemorrhagic shock. Methods Fifty Wistar rats were bled to establish the shock model and subsequently resuscitated with shed blood and normal saline. Just prior to resuscitation, the rats were randomly assigned to 5 groups： sham group, shock group, shock ＋ glycine group, shock ＋ MP group and shock ＋ glycine ＋ MP group. The intracellular calcium concentration and the level of tumor necrosis factor alpha （TNF alpha） in the culture medium of Kupffer cells were determined after stimulation with different concerntrations （1, 10, 100 and 1000 ng/ml） of lipopolysaccharide （LPS）. Results Concentration of intracellular calcium and production of TNF-α by isolated Kupffer cells stimulated by LPS were elevated significantly in the rats with hemorrhagic shock, which were totally prevented by glycine ＋ MP compared with other groups （ P 〈 0. 005 ）. Conclusions The combination of glycine and MP prevents the increase of intracellular calcium of Kupffer cell, suppress Kupffer cell activation, decrease the production of TNF-α of Kupffer cell and block systemic inflammatory responses more effectively than single administer of glycine or MP.