人尿激肽原酶通过调控NLRP3炎性小体对小鼠脑缺血/再灌注损伤的影响  

Effect of Human Urinary Kallindinogenase on cerebral ischemia/reperfusion injury through downregulating NLRP3 inflammasome in mice

作　　者：韩冬[1] 温璐璐[1] 王珏[1] 高岩[1] 冯娟[1] HAN Dong;WEN Lulu;WANG Jue(The First Department of Neurology,Shengjing Hospital of China Medical University,Shenyang 110004,China)

机构地区：[1]中国医科大学附属盛京医院第一神经内科,辽宁沈阳110004

出　　处：《中风与神经疾病杂志》2020年第7期629-632,共4页Journal of Apoplexy and Nervous Diseases

基　　金：辽宁省自然科学基金计划重点项目(No.20170541053)。

摘　　要：目的观察人尿激肽原酶对小鼠脑缺血/再灌注损伤的治疗作用,探讨人尿激肽原酶对脑缺血/再灌注损伤的保护机制。方法线栓法制备小鼠大脑中动脉闭塞(MCAO)模型,1 h后复灌,制备脑缺血/再灌注模型。人尿激肽原酶高剂量组(35×10^-3 PNAU/kg)、低剂量组(17.5×10^-3 PNAU/kg)分别于复灌后0.5 h静脉给药。TTC法测定梗死面积,Bederson评分法评定小鼠行为学,Elisa法测定血清、脑组织匀浆IL-1β、IL-18含量,Real time PCR、Western blot法测定脑组织NLRP3、ASC、caspase-1 mRNA及蛋白表达。结果人尿激肽原酶治疗给药可显著减轻小鼠脑缺血/再灌注损伤后的梗死面积,改善神经行为学评分。与假手术组比较,模型组小鼠NLRP3、ASC及caspase-1 mRNA及蛋白表达明显上调,血清及脑组织匀浆中IL-1β、IL-18表达增加;人尿激肽原酶治疗给药可显著抑制IL-1β、IL-18表达,下调NLRP3、ASC及caspase-1 mRNA及蛋白表达。结论人尿激肽原酶可能通过抑制小鼠脑缺血/再灌注后NLRP3炎性小体表达,进而抑制炎症反应,减轻脑缺血损伤。Objective To study the protective effect and possible mechanism of Human Urinary Kallindinogenase(UK)after ischemia-reperfusion injury in mice.Methods Middle cerebral artery occlusion(MCAO)was establish for 1 h and then reperfusion to mimic ischemic stroke in mice.UK treatment with high(35×10^-3PNAU/kg)and low(17.5×10^-3 PNAU/kg)does were intravenous injection after reperfusion for 30 min,respectively.2,3,5-triphenyltetrazolium chloride(TTC)staining was used to assay cerebral infarction size,neurological assessment was performed according to Bederson method.Elisa methods was applied to determine serum and brain tissue IL-1β、IL-18 content,and Real time PCR and Western blot were exerted to evaluate NLRP3,ASC,and caspase-1 mRNA and protein expression.Results The results showed that UK treatment attenuated infarction size in ischemic stroke mice,and improve neurological impairment.Compared with sham mice,NLRP3,ASC,and caspase-1 mRNA and protein expression were elevated,IL-1βand IL-18 concentration were increased obviously in MCAO/R mice.Human Urinary Kallindinogenase treatment inhibited IL-1βand IL-18 releasing,downregulating NLRP3,ASC,and caspase-1 mRNA and protein expression.Conclusion UK attenuates inflammatory response and infarction size partly through inhibiting NLRP3 inflammasome expression and cytokine releasing.

关 键 词：人尿激肽原酶 脑缺血/再灌注 NLRP3炎症小体 

分 类 号：R743.3[医药卫生—神经病学与精神病学]