胡黄连苷Ⅱ调节cAMP/PKA信号通路对缺氧缺血性脑损伤新生大鼠血脑屏障损伤的影响  

Effects of picroside II on blood-brain barrier injury in neonatal rats with hypoxic ischemic brain damage by modulating the cAMP/PKA signaling pathway

作　　者：关君艳 王静 GUAN Junyan;WANG Jing(Department of Pediatrics,Xiangyang Central Hospital,Affiliated Hospital of Hubei University of Arts and Science,Xiangyang 441000,Hubei,China)

机构地区：[1]湖北文理学院附属襄阳市中心医院儿科,襄阳441000

出　　处：《医学研究与战创伤救治》2024年第10期1027-1033,共7页Journal of Medical Research & Combat Trauma Care

摘　　要：目的探讨胡黄连苷Ⅱ(P-Ⅱ)通过调节cAMP/PKA信号通路对缺氧缺血性脑损伤(HIBD)新生大鼠血脑屏障(BBB)损伤的影响。方法建立HIBD模型,将新生大鼠按随机数字表法分为假手术组(Sham组)、模型组(HIBD组)、胡黄连苷Ⅱ低剂量组(P-Ⅱ-L)、胡黄连苷Ⅱ中剂量组(P-Ⅱ-M)、胡黄连苷Ⅱ高剂量组(P-Ⅱ-H)、胡黄连苷Ⅱ高剂量+PKA抑制剂组(P-ⅡH+H89),每组25只,检测脑含水量;ELLISA法检测MDA、ROS含量、NADPH活性和cAMP浓度;HE染色观察大脑组织形态;EB染色检测BBB通透性;DyLight 488荧光标记检测IgG泄露情况;Western blot检测Claudin-5、Occludin、VE-Cadherin、β-Catenin及p-PKA、PKA、p-CREB、CREB蛋白表达水平。结果HIBD组新生大鼠脑含水量、MDA、ROS含量、NADPH活性、IgG表达增加,Claudin-5、Occludin、VE-Cadherin、β-Catenin、cAMP浓度及p-PKA/PKA、p-CREB/CREB表达水平降低(P<0.05);与HIBD组比较,P-Ⅱ-L、P-Ⅱ-M、P-Ⅱ-H组新生大鼠脑含水量、MDA、ROS含量、NADPH活性、IgG呈剂量依赖性降低,Claudin-5、Occludin、VE-Cadherin、β-Catenin、cAMP浓度及p-PKA/PKA、p-CREB/CREB表达水平呈剂量依赖性增加(P<0.05);与P-Ⅱ-H组比较,P-Ⅱ-H+H89组脑含水量、MDA、ROS含量、NADPH活性、IgG表达增加,Claudin-5、Occludin、VE-Cadherin、β-Catenin、cAMP浓度及p-PKA/PKA、p-CREB/CREB表达水平降低(P<0.05)。结论P-Ⅱ通过激活cAMP/PKA信号通路保护HIBD新生大鼠免受BBB损伤。Objective To investigate the effects of picroside II(P-II)on blood-brain barrier(BBB)injury in neonatal rats with hypoxic-ischemic brain injury(HIBD)by modulating cAMP/PKA signaling pathway.Methods HIBD model was established.Neonatal rats were divided into sham surgery group(Sham group),model group(HIBD group),low dose picrosideⅡgroup(P-II-L),medium dose picrosideⅡgroup(P-II-M),high dose picrosideⅡgroup(P-II-H),high dose picrosideⅡ+PKA inhibitor group(P-IIH+H89)according to random number table method.Brain water content of 25 subjects in each group was detected.The content of MDA,ROS,NADPH activity and cAMP concentration were detected by ELLISA method.HE staining was used to observe brain morphology.The BBB permeability was detected by EB staining.DyLight 488 fluorescent marker was used to detect IgG leakage.Western blot was applied to detect the expression levels of Claudin-5,Occludin,VE-Cadherin,β-Catenin,and p-PKA,PKA,p-CREB,and CREB proteins.Results Brain water content,MDA,ROS content,NADPH activity and IgG expression were increased in HIBD group.The concentration of Claudin-5,Occludin,VE-Cadherin,β-Catenin,cAMP and the expression levels of P-PKA/PKA and P-CREB/CREB were decreased(P<0.05);Compared with HIBD group,brain water content,MDA,ROS content,NADPH activity and IgG of neonatal rats in P-II-L,PII-M and P-II-H groups were decreased in a dose-dependent manner.The concentrations of Claudin-5,Occludin,VE-Cadherin,β-Catenin,cAMP and the expression levels of P-PKA/PKA and P-CREB/CREB were increased in a dose-dependent manner(P<0.05).Compared with P-II-H group,brain water content,MDA,ROS content,NADPH activity and IgG expression were increased in P-II-H+H89 group.The concentration of Claudin-5,Occludin,VE-Cadherin,β-Catenin,cAMP and the expression levels of P-PKA/PKA and PCREB/CREB were decreased(P<0.05).Conclusion P-II protects HIBD neonatal rats from BBB injury by activating the cAMP/PKA signaling pathway.

关 键 词：胡黄连苷Ⅱ 环磷酸腺苷/蛋白激酶A 缺氧缺血性脑损伤 血脑屏障损伤 

分 类 号：R722[医药卫生—儿科]