小豆蔻明调节YAP/TAZ信号通路对脑梗死大鼠模型血脑屏障的影响
Effects of cardamomine on blood-brain barrier in rat model of cerebral infarction by regulating YAP/TAZ signaling pathway
张秋萍 1杨月君 2张彩丽3
作者信息
- 1. 071051 河北省保定市第二医院门诊部
- 2. 071051 河北省保定市第二医院神经内科
- 3. 071051 河北省保定市第二医院妇产科
- 折叠
摘要
目的 探讨小豆蔻明(Cardamonin,CAR)对脑梗死大鼠模型血脑屏障(Blood-brain barrier,BBB)及 Hippo-p-Yes 相关蛋白(Yes associated protein,YAP)/转录共激活因子(Tafazzin,TAZ)信号通路的影响.方法 建立脑梗死大鼠模型,所有大鼠分为对照组[Control(CT)组]、[模型组(Model(M)组]、CAR低剂量组(CAR L 组,5 mg·kg-1·d-1 CAR)、CAR 高剂量组(CAR H 组,20 mg·kg-1·d-1 CAR)和 CAR 高剂量+PY60组(CAR H+PY60组,20 mg·kg-1·d-1 CAR+10 mg/mL PY60);给药期间各组大鼠做神经功能评分;给药结束后苏木精-伊红(Hematoxylin-eosin,HE)染色观察脑组织病理变化,氯化三苯基四氮唑(Triphe-nyl-tetrazolium chloride,TCC)染色测定脑梗死体积,免疫组化检测脑组织闭合蛋白(Occludin)、紧密连接蛋白(Claudin-5)的表达水平,计算脑含水量和伊文思蓝(Evans blue,EB)渗透量,Western blot技术验证脑组织磷酸化(Phosphorylation,p)-哺乳动物 STE20 样蛋白激酶 1(Mammalian sterile20-like kinase1,MST1)、MST1、p-大肿瘤抑制因子 1(Large tumor suppressor factor 1,LATS1)、LATS1,p-YAP,YAP 蛋白表达水平.结果 与CT组比较,M组大鼠脑组织细胞间隙变大且有大量炎症细胞浸润及少量空泡变性,神经功能评分、脑梗死体积、脑含水量、EB渗透量及p-YAP/YAP蛋白表达水平升高,Occludin,Claudin-5,p-MST1/MST1,p-LATS1/LATS1蛋白表达水平降低(P＜0.05);与M组比较,CAR L,H组大鼠炎症细胞浸润、细胞空泡变性、细胞间隙减少,脑组织结构更完整,神经功能评分、脑梗死体积、脑含水量、EB渗透量及p-YAP/YAP蛋白表达水平降低,Occludin,Claudin-5,p-MST1/MST1,p-LATS1/LATS1 蛋白表达水平升高(P＜0.05);与CAR H组比较,CAR H+PY60组大鼠炎症细胞浸润和细胞空泡变性增加,神经功能评分、脑梗死体积、脑含水量、EB 渗透量及 p-YAP/YAP 蛋白表达水平升高,Occludin,Claudin-5,p-MST1/MST1,p-LATS1/LATS1蛋白表达水平降低(P＜0.05).结论 CAR可能通过抑制YAP/TAZ信号通路来改善脑梗死大鼠模型BBB功能.
Abstract
Objective To investigate the effects of cardamonin(CAR)on the blood-brain barrier(BBB)and the Hippo/p-Yes-associated protein(YAP)/transcription coactivator(TAZ)signaling pathway in a rat model of cerebral infarction.Methods A rat model of cerebral infarction was established,and all rats were di-vided into a control group(CT group),a model group(M group),a low-dose CAR group(CAR-L group,5 mg/kg/d CAR),a high-dose CAR group(CAR-H group,20 mg/kg/d CAR),and a high-dose CAR+PY60 group(CAR-H+PY60 group,20 mg/kg/d CAR+10 mg/m PY60);during the administration period,neuro-logical function scores were evaluated for each group.After administration,HE staining was applied to ob-serve pathological changes in brain tissue,triphenyltetrazolium chloride(TCC)staining was used to determine the volume of cerebral infarction,and immunohistochemistry was used to detect the expression levels of Occlu-din and Claudin-5 in brain tissue.The brain water content and Evans blue(EB)penetration were calculated.Western blotting was used to verify the expression levels of the brain tissue phosphorylation(p)proteins,mammalian STE20-like protein kinase 1(MST1),MST1,p-large tumor suppressor factor 1(LATS1),LATS1,p-YAP,and YAP.Results Compared with those in the CT group,the intercellular spaces in the brain tissue of rats in the M group were greater,with a large amount of inflammatory cell infiltration and a small amount of vacuolar degeneration.The neurological function score,cerebral infarction volume,brain wa-ter content,EB permeability,and p-YAP/YAP protein expression level increased,and the protein expression levels of Occludin,Claudin-5,p-MST1/MST1,and p-LATS1/LATS1 decreased(P＜0.05).Compared with those in the M group,the infiltration of inflammatory cells,cell vacuolar degeneration,and intercellular spaces in the CARL and H groups were lower,the brain tissue structure was more complete,the neurological func-tion score,cerebral infarction volume,brain water content,EB permeability,and p-YAP/YAP protein expres-sion levels were lower,and the protein expression levels of Occludin,Claudin-5,p-MST1/MST1,and p-LATS1/LATS1 were greater(P＜0.05).Compared with those in the CAR H group,the infiltration of in-flammatory cells and cellular vacuolar degeneration in the CAR H+PY60 group were greater,the neurological function score,cerebral infarction volume,brain water content,EB permeability,and p-YAP/YAP protein expression were greater,and the expression of Occludin,Claudin-5,p-MST1/MST1,and p-LATS1/LATS1 proteins was lower(P＜0.05).Conclusion CAR may improve BBB function in a rat model of cerebral infarc-tion by inhibiting the YAP/TAZ signaling pathway.
关键词
小豆蔻明/脑梗死/血脑屏障/Hippo/p-Yes/转录共激活因子信号通路Key words
Cardamonin/Cerebral infarction/Blood-brain barrier/Hippo/p-Yes/transcription coacti-vator signaling pathway引用本文复制引用
出版年
2024
NETL
NSTL
万方数据