目的

初步探讨巨噬细胞移动抑制因子（MIF）在大鼠急性坏死性胰腺炎（ANP）发病中的作用。

方法

健康雄性SD大鼠60只，随机分成三组，分别为对照组（腹腔注射生理盐水）、ANP组（腹腔注射左旋精氨酸）和干预组（腹腔注射左旋精氨酸＋单克隆抗MIF抗体），每组20只。采用左旋精氨酸改良方法建立ANP模型，分别于3、6、12、24 h四个不同时点处死5只大鼠，剖腹后从肠系膜上静脉取血，ELISA法测定血清MIF、TNF-α、IL-1、IL-6和IL-8水平；碘比法测定血淀粉酶；切取胰腺组织依据Kusske标准行胰腺病理学评分；Western blot法测定胰腺组织NF-κB p65蛋白的表达。

结果

ANP组血淀粉酶及胰腺组织病理学评分各时点与对照组相比均显著升高，干预组与ANP组相比均显著降低（P＜0.01）；胰腺组织NF-κB p65蛋白的表达于造模后3 h开始呈持续性上调，与对照组相比其表达显著增多，在干预组其表达水平与ANP组相比明显下调（P＜0.01）；血清MIF、TNF-α、IL-1、IL-6各时点在ANP组与正常对照组相比其水平均有明显升高，在干预组其水平与ANP组相比显著降低（P＜0.05）；血清IL-8在6、12、24 h不同时点水平变化同上述因子，但在3 h时点其水平无明显升高；MIF、胰腺组织病理学评分及胰腺组织NF-κB p65蛋白的表达，两两之间均成正相关（P＜0.05）。

结论

腹腔注射左旋精氨酸建立ANP的模型是稳定的；MIF可能通过调控核因子NF-κB的途径影响血清TNF-α、IL-1、IL-6及IL-8水平而在大鼠急性胰腺炎发病过程中起一定的作用。

Objective

To explore the role of macrophage migration inhibitory factor (MIF)in rat with acute necrotizing pancreatitis (ANP).

Methods

Sixty healthy male SD rats were randomly divided into normal control group (injected intraperitoneally with saline),ANP group (injected intraperitoneally with L-arginine)and intervention group (injected intraperitoneally with L-arginine and anti-MIF),20 mice in each group. Rats were killed 3, 6, 12, 24 h later respectively. Blood and pancreas tissues were obtained. The levels of serum TNF-α,MIF, IL-1, IL-6, IL-8 were determined with ELISA. Blood amylase was detected through indo-amylon chromatometry.Resected pancreas tissues were histopathological graded according to the standard of Kusske.The expression of NF-κB p65 protein in pancreatic tissues was performed by the way of Western blot.

Results

Compared to the normal control group,blood amylase and histopathological scores were significantly increased in ANP group,P＜0.01.The concentration of NF-κB in pancreatic tissues was significantly up-regulated in ANP group.However,it was significantly down-regulated in anti-MIF intervention group,P＜0.01.The levels of serum MIF,TNF-α,IL-1,IL-6 were significantly increased 3,6,12,24 h later.But the levels were significantly depressed in intervention group,P＜0.05.The range of the serum IL-8 at 6,12,24 h was similar with the cytokine described above.But there was no significantly different at 3 h with paired comparison. Positive correlation was found between histopathological scores,serum levels of MIF and the concentration of NF-κB in pancreatic tissues.

Conclusions

The rat models injected intraperitoneally with L-arginine are stable.Perhaps MIF has influence on the levelofserum TNF-α,IL-1,IL-6 and IL-8 by controlling NF-κB pathway.