Shriners Hospitals for Children, Galveston, Texas 77550, USA.
Shock. 2010 Mar;33(3):299-305. doi: 10.1097/SHK.0b013e3181b2f439.
Insulin resistance with its associated hyperglycemias represents one significant contributor to mortality in burned patients. A variety of cellular stress-signaling pathways are activated as a consequence of burn. A key player in the cellular stress response is the endoplasmic reticulum (ER). Here, we investigated a possible role for ER-stress pathways in the progression of insulin function dysregulation postburn. Rats received a 60% total body surface area thermal injury, and a laparotomy was performed at 24, 72, and 192 h postburn. Liver was harvested before and 1 min after insulin injection (1 IU/kg) into the portal vein, and expression patterns of various proteins known to be involved in insulin and ER-stress signaling were determined by Western blotting. mRNA expression of glucose-6-phosphatase and glucokinase were determined by reverse-transcriptase-polymerase chain reaction and fasting serum glucose and insulin levels by standard enzymatic and enzyme-linked immunosorbent assay techniques, respectively. Insulin resistance indicated by increased glucose and insulin levels occurred starting 24 h postburn. Burn injury resulted in activation of ER stress pathways, reflected by significantly increased accumulation of phospho-PKR-like ER-kinase and phosphorylated inositol requiring enzyme 1, leading to an elevation of phospho-c-Jun N-terminal kinase and serine phosphorylation of insulin receptor substrate (IRS) 1 postburn. Insulin administration caused a significant increase in tyrosine phosphorylation of IRS-1, leading to activation of the phosphatidylinositol 3 kinase/Akt pathway in normal liver. Postburn tyrosine phosphorylation of IRS-1 was significantly impaired, associated with an inactivation of signaling molecules acting downstream of IRS-1, leading to significantly elevated transcription of glucose-6-phosphatase and significantly decreased mRNA expression of glucokinase. Activation of ER-stress signaling cascades may explain metabolic abnormalities involving insulin action after burn.
胰岛素抵抗及其相关的高血糖是烧伤患者死亡的一个重要原因。烧伤会激活多种细胞应激信号通路。内质网(ER)是细胞应激反应的关键参与者。在这里,我们研究了 ER 应激途径在烧伤后胰岛素功能失调进展中的可能作用。大鼠接受 60%的全身体表面积热损伤,在烧伤后 24、72 和 192 小时进行剖腹手术。在门静脉内注射胰岛素(1IU/kg)前和 1 分钟后采集肝脏,并通过 Western blot 测定已知参与胰岛素和 ER 应激信号的各种蛋白质的表达模式。通过逆转录-聚合酶链反应测定葡萄糖-6-磷酸酶和葡糖激酶的 mRNA 表达,通过标准酶和酶联免疫吸附测定技术分别测定空腹血清葡萄糖和胰岛素水平。烧伤后 24 小时开始出现胰岛素抵抗,表现为血糖和胰岛素水平升高。烧伤导致 ER 应激途径的激活,表现为磷酸化 PKR 样 ER 激酶和磷酸化肌醇需要酶 1 的显著增加,导致磷酸化 c-Jun N-末端激酶和胰岛素受体底物(IRS)1 的丝氨酸磷酸化增加。胰岛素给药导致 IRS-1 的酪氨酸磷酸化显著增加,导致正常肝脏中磷酸肌醇 3 激酶/Akt 途径的激活。烧伤后 IRS-1 的酪氨酸磷酸化显著受损,与 IRS-1 下游信号分子的失活相关,导致葡萄糖-6-磷酸酶的转录显著升高,葡糖激酶的 mRNA 表达显著降低。ER 应激信号级联的激活可以解释烧伤后涉及胰岛素作用的代谢异常。