基于JAK2/STAT1信号通路探讨骨桥蛋白缓解高血糖大鼠脑出血后炎症反应机制  

To investigate the mechanism of osteopontin alleviating inflammatory response after cerebral hemorrhage in hyperglycemia rats based on JAK2/STAT1 signaling pathway

作　　者：郝雨知 蔡啸 金一峰 孟玉娟 李天娇 Hao Yuzhi;Cai Xiao;Jin Yifeng(Department of Pharmacy,Shenyang Hospital of Traditional Chinese Medicine,Shenyang Liaoning 110004;不详)

机构地区：[1]沈阳市中医院药剂科,110004 [2]辽宁中医药大学附属医院药剂科 [3]北部战区总医院药剂科

出　　处：《卒中与神经疾病》2021年第6期622-630,共9页Stroke and Nervous Diseases

基　　金：辽宁省自然科学基金指导计划项目(20170217)。

摘　　要：目的基于非受体酪氨酸激酶2/信号转导与转录激活因子1(Just another kinase 2/Signal transducer and activator of transcription 1,JAK2/STAT1)信号通路探讨骨桥蛋白(Osteopontin, OPN)缓解高血糖大鼠脑出血(Intracerebral hemorrhage, ICH)后炎症反应机制。方法采用胶原酶注射法建立脑出血大鼠模型,ICH后第3 h腹腔注射50%葡萄糖诱发急性脑血肿扩张;ICH后第1 h鼻腔给药重组骨桥蛋白(Recombinant osteopotin, rOPN);采用脑组织水含量(Brain water content, BWC)、神经功能缺损评分、蛋白免疫印迹试验和免疫组化等方法评价脑损伤的发生发展;通过整合蛋白β1小干扰RNA(Small interfering RNA,siRNA)和JAK2激动剂香豆霉素(Coumermycin A1,C-A1)对其分子机制进行研究。结果 ICH后第24 h OPN表达水平显著下降,然后在第48 h显著上调并持续至第72 h;整合蛋白β1也呈同样表达模式。此外,在ICH后第24 h磷酸化-非受体酪氨酸激酶2/非受体酪氨酸激酶2(Phosphorylation-Janus kinase 2/Janus kinase 2,p-JAK2/JAK2、磷酸化-信号转导与转录激活因子1/信号转导与转录激活因子1(Phosphorylation-signal transducer and activator of transcription 1/Signal transducer and activator of transcription 1,p-STAT1/STAT1)的比值均显著升高。低剂量的OPN对脑组织含水量(BWC)无明显影响,而中、高剂量的OPN均能显著降低ICH诱导的BWC升高;行为学评分显示可改善神经功能,但高剂量和中剂量的OPN在效果上没有明显差异;OPN可减弱BWC的增加,从而改善ICH诱导的神经功能障碍;中剂量rOPN(3μg)和高剂量rOPN(9μg)可使脑组织OPN表达水平升高,脑组织中OPN的增加伴随着整合蛋白β1的增多;ICH诱导p-JAK2/JAK2、p-STAT1/STAT1比值增高,而OPN的增加伴随着p-JAK2/JAK2、p-STAT1/STAT1比值的降低,ICH诱导肿瘤坏死因子a(Tumor necrosis factor, TNF-a)和白细胞介素-1β(Interleukin-1β,IL-1β)的过量产生,并伴有基质金属蛋白酶-9(Matrix metallopeptidase 9,MMP-9)的激�Objective To investigate the mechanism of osteopontin(OPN) alleviating inflammatory response after hyperglycemia ICH in rats based on the JAK2/STAT1 signaling pathway. Methods Intracerebral hemorrhage rat model was established by collagenase injection. Acute cerebral hematoma dilation was induced by intraperitoneal injection of 50% glucose 3 h after ICH. Recombinant osteopotin(rOPN) was administered intranasally 1 h after ICH. Brain water content(BWC), neurological deficit, Western blot and immunohistochemistry were used to evaluate the occurrence and development of brain injury. The molecular mechanisms and pathways of integrin β1 siRNA and JAK2 agonist C-A1 were studied. Results OPN expression was significantly decreased at 24 h after ICH, and then continued to increase at 48-72 h. Integrin β1 also showed the similar expression pattern. In addition, the ratios of p-JAK2/JAK2 and p-STAT1/STAT1 were significantly increased 24 h after ICH. Low doses of OPN had no effect on brain water content(BWC), while both medium and high doses of OPN could significantly inhibit ICH induced BMC increasion. Behavioral scores showed improvement of neurological function, but there was no significant difference between high and medium doses of OPN. OPN attenuated the increase of BWC and improved the neurologic dysfunction induced by ICH. The expression of OPN in brain tissue was increased by medium dose(3 μg) and high dose(9 μg) rOPN treatment, accompanied by the increase of integrin β1 expression. ICH induced the increase of p-JAK2/JAK2 and p-STAT1/STAT1 ratios, while the increase of OPN was accompanied by the decrease of p-JAK2/JAK2 and p-STAT1/STAT1 ratios. ICH induced excessive production of TNF-a and IL-1β, accompanied by activation of MMP-9, while rOPN-induced inhibition of the JAK2/STAT1 pathway resulted in decreased production of TNF-a and IL-1β, leading to a dose-dependent reduction of ICH-induced MMP-9 overexpression. In addition, immunohistochemistry staining of neutrophil specific protein MPO and Western blotting

关 键 词：非受体酪氨酸激酶2 信号转导与转录激活因子1 骨桥蛋白 脑出血 炎症反应 

分 类 号：R743.34[医药卫生—神经病学与精神病学]