双氢青蒿素调节cAMP/PKA/CREB信号通路对缺血性脑卒中大鼠神经炎症和血脑屏障的影响  

Impacts of dihydroartemisinin on neuroinflammation and blood⁃brain barrier in ischemic stroke rats by regulating the cAMP/PKA/CREB signaling pathway

作　　者：黄一苇 方建 唐森 HUANG Yiwei;FANG Jian;TANG Sen(Department of Neurology,Zhumadian Central Hospital,Zhumadian,Henan 463000,China)

机构地区：[1]河南省驻马店市中心医院神经内科,河南驻马店463000

出　　处：《热带医学杂志》2025年第1期39-43,F0004,共6页Journal of Tropical Medicine

基　　金：河南省医学科技攻关计划项目(LHGJ20210555)。

摘　　要：目的探讨双氢青蒿素对缺血性脑卒中(IS)大鼠神经炎症和血脑屏障(BBB)的影响以及其机制。方法通过大脑中动脉阻塞术制备IS大鼠模型,将造模成功大鼠分为模型组、双氢青蒿素组、双氢青蒿素+H-89(蛋白激酶A抑制剂)组,每组20只,另取20只大鼠设置为假手术组;药物干预3 d后,使用神经功能损伤评分(mNSS)评价各组大鼠神经功能损伤程度;氯化三苯基四氮唑(TTC)染色检测脑梗死体积;干湿重法检测脑含水量;分光光度计法检测各组大鼠脑组织伊文氏蓝含量;ELISA检测大鼠脑组织白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、IL-6、环磷酸腺苷(cAMP)水平;Western blot检测大鼠脑组织中蛋白激酶A催化亚基(PKAc)、cAMP反应元件结合蛋白(CREB)、磷酸化CREB(p-CREB)表达。结果与假手术组比较,模型组大鼠mNSS评分、脑梗死体积、脑含水量、脑组织伊文氏蓝含量,脑组织IL-1β、TNF-α、IL-6水平显著升高,脑组织cAMP含量、PKAc、p-CREB/CREB表达显著降低,差异均有统计学意义(q=51.730、70.318、6.420、45.407、16.306、24.651、13.570、18.197、21.236、21.609,P均<0.05)。与模型组比较,双氢青蒿素组大鼠mNSS评分、脑梗死体积、脑含水量、脑组织伊文氏蓝含量、脑组织IL-1β、TNF-α、IL-6水平显著降低,脑组织cAMP含量、PKAc、p-CREB/CREB表达显著升高,差异均有统计学意义(q=21.545、30.107、5.690、35.506、15.158、18.359、11.216、12.955、14.816、14.749,P均<0.05)。与双氢青蒿素组比较,双氢青蒿素+H-89组大鼠mNSS评分、脑梗死体积、脑含水量、脑组织伊文氏蓝含量、脑组织IL-1β、TNF-α、IL-6水平显著升高,脑组织cAMP含量、PKAc、p-CREB/CREB表达显著降低,差异均有统计学意义(q=13.642、24.642、4.489、29.952、12.933、11.728、8.830、8.271、12.841、13.377,P均<0.05)。结论双氢青蒿素通过激活cAMP/PKA/CREB信号通路减轻IS大鼠神经炎症以及恢复BBB。Objective To explore the impacts of dihydroartemisinin on neuroinflammation and blood-brain barrier(BBB)in ischemic stroke(IS)rats and its mechanisms.Methods An IS rat model was prepared by middle cerebral artery occlusion surgery.The successfully modeled rats were grouped into a model group,a dihydroartemisinin group,and a dihydroartemisinin+H-89(protein kinase A inhibitor)group,with 20 rats in each group.Another 20 rats were selected as a sham operation group;after 3 days of drug intervention,the modified neurological severity score(mNSS)was applied to evaluate the degree of neurological damage of rats in each group;triphenyltetrazolium chloride(TTC)staining was applied to detect the volume of cerebral infarction;the dry wet weight method was applied to detect brain water content;spectrophotometer method was applied to detect the content of Evans blue in the brain tissue of rats in each group;ELISA was applied to detect the levels of interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),IL-6,and cyclic adenosine monophosphate(cAMP)in rat brain tissue;Western blot was applied to detect the expression of protein kinase A catalytic subunit(PKAc),cyclic-AMP response binding protein(CREB),and p-CREB in rat brain tissue.Results Compared with the sham surgery group,the mNSS score,cerebral infarction volume,brain water content,Evans blue content in brain tissue,and contents of IL-1β,TNF-α,and IL-6 in brain tissue of rats in the model group were obviously increased;the content of cAMP,the expression of PKAc and p-CREB/CREB in brain tissue were obviously reduced(q=51.730,70.318,6.420,45.407,16.306,24.651,13.570,18.197,21.236,21.609;all P<0.05).Compared with the model group,the mNSS score,cerebral infarction volume,brain water content,Evans blue content in brain tissue,and contents of IL-1β,TNF-α,and IL-6 in brain tissue of rats in the dihydroartemisinin group were obviously reduced;the content of cAMP,the expression of PKAc and p-CREB/CREB in brain tissue were obviously increased(q=21.545,30.107,5.690,35.506,15.15

关 键 词：双氢青蒿素 cAMP/PKA/CREB 缺血性脑卒中 神经炎症 血脑屏障 

分 类 号：R743.3[医药卫生—神经病学与精神病学]