烧伤血清促进大鼠库普弗细胞分泌肿瘤坏死因子α

The postburn serum stimulates the secretion of TNFα by Kupffer cells

目的：探讨烫伤血清对库普弗细胞（ＫＣ）分泌肿瘤坏死因子α（ＴＮＦ－α）作用及机制。方法：胶原酶肝脏灌注消化、梯度离心分离培养ＫＣ，用烫伤血清刺激并加入甘氨酸和尼莫地平，用双抗体夹心酶联免疫法测定培养上清中ＴＮＦ－α的含量。结果：加入２０％烫伤血清后的ＫＣ培养上清内ＴＮＦα含量明显增加，高峰在２ｈ；加入甘氨酸（１ｍｍｏｌ／Ｌ）和尼莫地平（１μｍｏｌ／Ｌ）后，培养上清内ＴＮＦ－α含量明显下降，在２ｈ仅为烫伤血清组的３５．１％和２７．６％。结论：烫伤血清可以刺激体外培养的ＫＣ分泌ＴＮＦ－α，其作用是经细胞内钙依赖的途径实现的。

Objective: To investigate the effect of postburn serum on the secretion of TNFα by Kupffer cells(KC) and its mechanism. Methods: KC were isolated by collagenase Ⅰ using portal perfusion and gradient centrifugation; KC were cultured in DMEM with 20% postburn serum, some of which were added with glycine(1 mmol/L)or nimodipine(1 μmol/L). Then their supernants were collected to detect the production of TNFα with ELISA kits. Results: The level of TNFα in the supernants of cultured KC with 20% postburn serum increased significantly. Its peak was at 2 h after culture. After adding glycine or nimodipine, the level of TNFα declined significantly. At 2 h after treatment, the production of TNFα was only 35.1% and 27.6% of that in no treatment group, respectively. Conclusion: Postburn serum can stimulate the secretion of TNFα by KC, which is dependent on intracellular calcium. 