p38丝裂原活化蛋白激酶参与对失血性和内毒素性休克耐受性的诱导。

Involvement of p38 mitogen-activated protein kinase in the induction of tolerance to hemorrhagic and endotoxic shock.

作者信息

Mendez C, Jaffray C, Wong V, Salhab K F, Kramer A A, Carey L C, Norman J G

机构信息

Department of Surgery, University of South Florida, Tampa, Florida 33612, USA.

出版信息

J Surg Res. 2000 Jun 15;91(2):165-70. doi: 10.1006/jsre.2000.5936.

DOI:10.1006/jsre.2000.5936

PMID:10839967

Abstract

BACKGROUND

Exposure to sublethal hemorrhage (SLH) makes rats tolerant to subsequent hemorrhagic or septic shock and is associated with altered NF-kappaB activity. The purpose of this study was to explore whether changes in p38 mitogen-activated protein (MAP) kinase activity also occur in the induction of tolerance by SLH.

METHODS

Rats were made tolerant by SLH or sham operation. Twenty-four hours later rats were exposed to lipopolysaccharide (LPS) or had peritoneal macrophages (Mphi) isolated. CNI-1493, a p38 MAP kinase inhibitor, or saline was given prior to SLH. Lungs were harvested 1 h after SLH or LPS and total protein was extracted. Peritoneal Mphi were stimulated with LPS (10 microg/ml) and total protein was isolated 1 h later. Active, dually phosphorylated p38 MAP kinase was determined by Western blot. Tumor necrosis factor (TNF) was measured in Mphi supernatants by enzyme-linked immunosorbent assay (ELISA) 18 h after LPS.

RESULTS

SLH activated p38 MAP kinase in the lung and this was inhibited by CNI-1493. Twenty-four hours later, lung p38 MAP kinase activity increased to the same degree in tolerant and sham rats following LPS, but much more prominently in the CNI-1493 treated rats. There was no p38 activity in peritoneal Mphi at baseline, and similar to lung p38, LPS led to increased p38 activity which was most significant in Mphi from rats that received CNI-1493 prior to SLH. TNF production by tolerant Mphi in response to LPS was significantly (P < 0.05, t test) decreased and p38 inhibition with CNI-1493 at the time of SLH reversed the inhibitory effects of tolerance on TNF production.

CONCLUSIONS

TNF production by tolerant Mphi following a second insult (LPS) is attenuated despite preservation of normal p38 MAP kinase activity. However, activation of this intracellular second messenger is a necessary step in the "cellular reprogramming" that occurs during the induction of tolerance by SLH.

摘要

背景

暴露于亚致死性出血（SLH）可使大鼠对随后的出血性或脓毒性休克产生耐受性，且与核因子κB（NF-κB）活性改变有关。本研究的目的是探讨在SLH诱导耐受性过程中，p38丝裂原活化蛋白（MAP）激酶活性是否也发生变化。

方法

通过SLH或假手术使大鼠产生耐受性。24小时后，将大鼠暴露于脂多糖（LPS）或分离腹腔巨噬细胞（Mphi）。在SLH之前给予p38 MAP激酶抑制剂CNI-1493或生理盐水。在SLH或LPS后1小时采集肺组织并提取总蛋白。用LPS（10微克/毫升）刺激腹腔Mphi，1小时后分离总蛋白。通过蛋白质免疫印迹法测定活性双磷酸化p38 MAP激酶。在LPS刺激18小时后，通过酶联免疫吸附测定法（ELISA）测量Mphi上清液中的肿瘤坏死因子（TNF）。

结果

SLH激活了肺组织中的p38 MAP激酶，而CNI-1493可抑制这种激活。24小时后，LPS刺激后，耐受性大鼠和假手术大鼠肺组织中的p38 MAP激酶活性升高程度相同，但在接受CNI-1493治疗的大鼠中升高更为显著。基线时腹腔Mphi中无p38活性，与肺组织中的p38相似，LPS导致p38活性增加，这在SLH前接受CNI-1493治疗的大鼠的Mphi中最为显著。耐受性Mphi对LPS刺激产生的TNF显著减少（P<0.05，t检验），SLH时用CNI-1493抑制p38可逆转耐受性对TNF产生的抑制作用。