芦荟素抑制p65的磷酸化对大鼠脑缺血再灌注诱导的组织损伤和炎症的调节作用  

Regulatory effects of Aloin inhibiting p65 phosphorylation on tissue damage and inflammation induced by rats with cerebral ischemia-reperfusion injury

作　　者：赵杨[1] 孙波 杜书章[2] ZHAO Yang;SUN Bo;DU Shu-Zhang(Department of Pharmacy,The Fifth Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,China)

机构地区：[1]郑州大学第五附属医院药学部,郑州450000 [2]郑州大学第一附属医院药学部,郑州450000

出　　处：《中国免疫学杂志》2019年第17期2084-2088,2093,共6页Chinese Journal of Immunology

基　　金：河南省医学科技攻关基金资助项目(201702119)

摘　　要：目的:探讨芦荟素(Aloin)抑制p65的磷酸化对大鼠脑缺血再灌注(CIRI)诱导的组织损伤和炎症的调节作用。方法:将64只SD大鼠随机分为对照组、Aloin组、MCAO组和Aloin-MCAO组各16只,MCAO组和Aloin-MCAO组参照改良Longa法制备动物脑中动脉缺血再灌注(MCAO)模型。Aloin组和Aloin-MCAO组腹腔注射Aloin50mg/kg,连续7d。治疗后,检测10d内大鼠存活率,对各组大鼠进行神经功能缺损评分,HE染色和TUNEL法检测各组大鼠脑组织损伤,采用免疫组化染色,检测脑组织细胞黏附分子-1(ICAM-1)的表达量;ELISA法测定脑组织中白介素(IL)-6、IL-10水平,Westernblot检测NF-κBp65的磷酸化、下游靶基因肿瘤坏死因子(TNF-)α以及凋亡标记分子Caspase-3和Caspase-9的表达。结果:与对照组相比,MCAO组的存活率显著下降(P<0.05),神经功能缺损评分、细胞凋亡率、ICAM-1、IL-6、p-NF-κBp65/NF-κBp65、TNF-α、Caspase-3和Caspase-9的表达均显著升高(P<0.05);与MCAO组相比,Aloin-MCAO组的存活率、IL-10显著升高(P<0.05),神经功能缺损评分、细胞凋亡率、ICAM-1、IL-6、p-NF-κBp65/NF-κBp65、TNF-α、Caspase-3和Caspase-9的表达均显著下降(P<0.05)。结论:Aloin可能通过抑制NF-κBp65信号通路抑制TNF-α的表达,降低机体炎症因子水平和细胞凋亡率,改善CIRI大鼠神经功能。Objective: To explore the regulatory effects of p65 phosphorylation inhibition by Aloin on tissue damage and inflammation induced by rats with cerebral ischemia-reperfusion injury (CIRI). Methods: 64 SD rats were randomly divided into control group,Aloin group,MCAO group and Aloin-MCAO group,with 16 rats in each group.MCAO group and Aloin-MCAO group prepared middle cerebral artery occlusion (MCAO) models according to modified Longa method.Aloin group and Aloin-MCAO group were intraperitoneally injected with 50 mg/kg Aloin for 7 consecutive days.After treatment,the survival rate was measured within 10 d,and the neurological deficit score was given in each group.HE staining and TUNEL method were used to detect brain tissue damage in each group.Immunohistochemical staining was used to detect the expression level of intercellular adhesion molecule-1 (ICAM-1).ELISA method was used to detect the levels of interleukin (IL)-6 and IL-10 in brain tissue,and Western blot was used to detect the NF-κB p65 phosphorylation,and expression of downstream target gene tumor necrosis factor (TNF-)α and apoptotic marker molecules Caspase-3 and Caspase-9. Results: Compared with control group,the survival rate in MCAO group was significantly decreased ( P <0.05),and neurological deficit score,apoptosis rate and expression levels of ICAM-1,IL-6,p-NF-κBp65/NF-κBp65,TNF-α,Caspase-3 and Caspase-9 were significantly increased ( P <0.05).Compared with MCAO group,the survival rate and IL-10 in Aloin-MCAO group were significantly increased ( P <0.05),and the neurological deficit score,apoptosis rate and expression levels of ICAM-1,IL-6,p-NF-κBp65/NF-κBp65,TNF-α,Caspase-3 and Caspase-9 were significantly decreased ( P <0.05). Conclusion: Aloin may inhibit the expression of TNF-α by inhibiting NF-κB p65 signaling pathway,reduce the inflammatory factors levels and apoptosis rate,and improve the neurological function of CIRI rats.

关 键 词：芦荟素 脑缺血再灌注 NF-ΚBP65 TNF-α 

分 类 号：R543[医药卫生—心血管疾病]