摘要
目的探讨JAK/STAT信号通路在胰弹性蛋白酶(elastase)诱导Kupffer细胞分泌促炎症因子中的作用机制。方法采用酶消化法和密度梯度离心法将提取的肝脏Kupffer细胞分为4组:A组(正常对照组,在培养基上清液中加入30μl/ml生理盐水);B组[用50ng/ml脂多糖(LPS)进行处理];C组(用50ng/ml LPS和1U/ml elastase进行处理);D组[用AG490(30nmol/ml)预先刺激0.5h后,再用LPS(50ng/ml)和elastase(1U/ml)处理]。采用酶免疫吸附(ELISA)法检测Kupffer细胞上清液中IL-6和TNF-α含量;Western blotting法检测细胞总蛋白中JAK2的表达情况。结果与A组比较,B组在给予LPS刺激后,JAK2的表达及上清液中IL-6和TNF-α的含量均明显增加(P<0.01);C组在同时给予LPS和elastase刺激后,JAK2的表达显著升高,上清液中IL-6和TNF-α含量与B组比较亦显著增加(P<0.01);而D组在预先给予AG490处理后,JAK2蛋白的表达明显下降,上清液中IL-6和TNF-α含量也有不同程度降低,与C组比较差异有统计学意义(P<0.01),但与B组比较,各值仅有轻微变化,差异无统计学意义(P>0.05)。结论抑制JAK/STAT通路的活化可下调elastase诱导Kupffer细胞分泌促炎症因子,有助于减轻急性胰腺炎时的炎症反应和肝损伤。
Objective To investigate the mechanism of JAK/STAT signal pathway in the secretion of proinflamnmtory cytokines by pancreatic elastase-induced rat Kupffer cells of rats with experimental acute pancreatids. Methods Hepatic Kupffer cells were isolated through enzymatic digestion and density gradient centrifugation, and then divided into four groups： group A （control group, with 30μl/ml normal saline in the supernatant）, group B （with 50ng/ml LPS）, group C （with 50ng/ml LPS and 1U/ml elastase） and group D （pretreated with 30 nmol/ml AG490 for 0. 5h, and then with 50ng/ml I.PS and 1U/ml elastase）. The concentrations of IL-6 and TNFa in Kupffer cell supernatant were determined by EI.ISA, and the expression of JAK2 protein was determined by Western blotting. Results The con centrations of IL-6, TNF-α in the supernatant and JAK2 protein in group B increased significantly compared with those in group A （ P〈0. 01）. The same result appeared when compared group C with those of group B （P〈0. 01）. The concentrations of IL-6, TNF-α and JAK2 protein in group D decreased significantly compared with those in group C （ P〈0. 01）, while only slightly increased compared with those in group B （ P〉0. 05）. Conclusion Inhibition of JAK/STAT signal pathway activation may reduce the pancreatic elastase-induced expression of proinflammatory cytokines in rat Kupffer cells, which may help alleviate the liver injury induced by acute pancreatitis.
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2008年第10期1167-1169,共3页
Medical Journal of Chinese People's Liberation Army
基金
南京军区南京总医院科研基金资助项目(2006020)
