KS-370G 對於ICR小鼠肝臟缺血再灌流的保護作用

KS-370G Alleviates Hepatic Ischemia/Reperfusion Injury in ICR mice

陳君怡(Chun-Yi Chen)

指導教授 ： 蘇銘嘉

國立臺灣大學/醫學院/藥理學研究所/碩士(2012年)

https://doi.org/10.6342/NTU.2012.10839

摘要

背景:在肝臟手術或是進行肝臟移植時，肝臟缺血再灌流是一個不可避免的傷害，並且因而造成肝臟本身功能不正常抑或是失去功能。到目前為止，並沒有一個主要的療法。目前，據我們所知，給予Caffeic acid phenyl ester (CAPE)在肝臟缺血再灌流的傷害中，可以發現改善肝臟組織的傷害以及肝指數，KS-370G本身是一個caffeamide的衍生物，結構類似Caffeic acid phenyl ester (CAPE)，但是在肝臟的作用，目前仍一無所知，故本篇研究目的在於評估KS-370G 是否能夠改善肝臟缺血再灌流所造成的肝功能損傷。
方法:我們在ICR小鼠在肝臟缺血前15分鐘，並且以尾靜脈注射方式注入1 mg/kg的劑量，之後再進行70%肝臟缺血約1個小時，接著實行3小時的再灌流。
結果: 經過肝臟再灌流3小時後，蒐集肝臟組織以及血液去評估KS-370G的保護作用。我們發現KS-370G 可以改善肝臟缺血再灌流引起肝功能不良的情形。並且肝臟再灌流3小時後，KS-370G 藉由活化PI3K-Akt路徑去誘導Bcl-2和Bcl-xl的活化並且同時抑制Bax的蛋白表現，若在給予KS-370G時，同時投予LY294002 (PI3K的抑制劑)時，會抑制KS-370G 對於肝臟損傷的保護程度以及增加細胞凋亡(apoptosis)的作用。此外，KS-370G 可以減少TNFα以及P38蛋白磷酸化並且同時也可以增加STAT和AMPK的活化。
結論: KS-370G 在肝臟缺血再灌流是具有保護作用，且歸功於活化細胞存活路徑PI3K-Akt路徑進而去抑制細胞凋亡作用，並且本身也有藉由抑制TNF-α和P38蛋白磷酸化的情形達到抗發炎的作用，以及(或)活化AMPK的作用。

關鍵字

PI3K-Akt ； caffeamide ； 肝臟缺血再灌流的損傷 ； 細胞凋亡

並列摘要

Purpose:
Hepatic ischemia-reperfusion injury is an unavoidable consequence which leads to primary liver dysfunction and organ failure during liver surgery and transplantation. There are at present no targets treatments against liver I/R damage. Recently，caffeic acid phenyl ester (CAPE) evidently attenuates
hepatic ischemia-reperfusion injury via decreasing histological damage score and the serum transaminase levels. The structure of KS-370G, a caffeamide derivative, is similar to CAPE. However, the effect of KS-370G on liver has not been studied. We aimed to study whether KS-370G could alleviate hepatic ischemia-reperfusion damage in livers from normal mice.
Materials and Methods:
We performed a 70% liver-ischemia (60 min) reperfusion model in ICR male mice. KS-370G was given at15 min (1 mg/kg, i.v.) before ischemia and the following 3 h reperfusion.
Results:
Liver tissues and blood were sampled at 3 h after reperfusion for detection of the protection efficacy of KS-370G. KS-370G strikingly ameliorated the deterioration of liver function in the mice after ischemia-reperfusion injury. PI3K-Akt pathway was activated at 3h after reperfusion. Besides, KS-370G conspicuously down-regulated Bax and accompanied with up-regulated Bcl-2 and Bcl-xl. Treatment with specific PI3k inhibitor LY294002 deteriorated I/R injury and elevated apoptotic effect despite KS-370G treatment. Moreover, KS-370G suppressed the increase in TNFα and phosphorylated p38 protein expression and increased STAT3 and AMPK phosphorylation.
Conclusion
In conclusion, KS-370G alleviates ischemia-reperfusion injury in liver by activation of cell survival PI3K-Akt signaling pathway to down-regulate apoptosis effect and anti-inflammatory effect by inhibition P38 activation and TNF-alpha production and/or activation of AMPK.

並列關鍵字

PI3K-Akt ； caffeamide ； hepatic ischemia-reperfusion injury ； apoptosis

參考文獻

【1】 Fondevila C, Busuttil RW, Kupiec-Weglinski JW. Hepatic ischemia/reperfusion injury – a fresh look. Exp Mol Pathol 2003;74:86–93.

【2】 Teoh NC, Farrell GC .Hepatic ischemia reperfusion injury: Pathogenic mechanisms and basis for hepatoprotection. J Gastroenterol Hepatol 2003;18:891–902

【3】 Jaeschke H, Bautista AP, Spolarics Z, Spitzer JJ. Superoxide generation by Kupffer cells and priming of neutrophils during reperfusion after hepatic ischemia. Free Radic Res Commun 1991;15:277–84.

【4】 Sindram D, Porte RJ, Hoffman MR, Bentley RC, Clavien PA. Platelets induce sinusoidal endothelial cell apoptosis upon reperfusion of the cold ischemic rat liver. Gastroenterology 2000; 118:183–91.

【5】 Samarasinghe DA, Tapner M, Farrell GC. Role of oxidative stress in hypoxia– reoxygenation injury to cultured rat hepatic sinusoidal endothelial cells. Hepatology 2000;31:160–5.