摘要
目的探讨嘌呤能受体2X4(P2X4)调控含NLR家族pyrin结构域蛋白1(NLRP1)/半胱氨酸天冬氨酸酶1(Caspase-1)通路在脑出血炎性损伤中的作用机制,为寻找脑出血治疗新措施提供实验依据。方法选取100只8~10周C57BL/6雄性小鼠建立脑出血模型,采用随机数字表法分为对照组、模型组、P2X4激动剂组、P2X4拮抗剂组、NLRP1激动剂组,每组各20只。对照组、模型组经小鼠经腹腔给予15μl生理盐水,P2X4激动剂组给予15μl胞苷5′-三磷酸(5′-CTP);P2X4拮抗剂组小鼠经腹腔注射15μl P2X4特异性拮抗剂5-BDBD;NLRP1激动剂组给予15μl胞壁酰二肽(MDP)。在实验第1、3、5、7天进行改良神经功能缺损评分(mNSS),采用ELISA法测量炎症因子[白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)]水平,采用蛋白免疫印迹法(Western blot)测定P2X4、NLRP1、Caspase-1蛋白表达,采用实时荧光定量聚合酶链式反应(qPCR)法测定P2X4、NLRP1、Caspase-1 mRNA表达,并进行组间比较。结果建模后第1、3、5、7天,模型组的mNSS评分高于对照组,P2X4激动剂组、NLRP1激动剂组的mNSS评分高于模型组,而P2X4拮抗剂组的mNSS评分低于模型组,差异有统计学意义(P<0.05)。建模后第1、3、5、7天,模型组的IL-1β、TNF-α水平高于对照组,P2X4激动剂组、NLRP1激动剂组的IL-1β、TNF-α高于模型组,而P2X4拮抗剂组的IL-1β、TNF-α低于模型组,差异有统计学意义(P<0.05)。模型组的P2X4、NLRP1、Caspase-1 mRNA和蛋白表达高于对照组,P2X4激动剂组、NLRP1激动剂组的相应指标均高于模型组,而P2X4拮抗剂组的相应指标低于模型组,差异有统计学意义(P<0.05)。结论P2X4通过调控NLRP1/Caspase-1通路参与脑出血炎性损伤的发生发展,抑制该通路对改善炎症反应有一定积极作用,值得深入研究。
Objective To explore the mechanism of purinergic receptor 2X4(P2X4)regulating NLR family pyrin domain-containing protein 1(NLRP1)/cystein-containing aspartat-specific protease-1(Caspase-1)pathway in the inflammatory injury of cerebral hemorrhage,and to provide experimental basis for finding new treatment measures for cerebral hemorrhage.Methods A total of 100 C57BL/6 male mice of 8-10 weeks were selected to establish cerebral hemorrhage model,and were divided into control group,model group,P2X4 agonist group,P2X4 antagonist group and NLRP1 agonist group according to random number table method,with 20 mice in each group.The control group and model group were given 15μl normal saline through the peritoneum,and the P2X4 agonist group was given 15μl cytidine 5'-triphosphate(5'-CTP).P2X4 antagonist group was intraperitoneally injected with 15μl of P2X4 specific antagonist 5-BDBD.The NLRP1 agonist group was given 15μl muramyl dipeptide(MDP).On 1,3,5 and 7 d,the modified neurological severityscores(mNSS)was performed,and the levels of inflammatory factors(interleukin-1β[IL-1β]and tumor necrosis factor-α[TNF-α])were measured by ELISA.The protein expressions of P2X4,NLRP1 and Caspase-1 were determined by Western blot,and the mRNA expressions of P2X4,NLRP1 and Caspase-1 were determined by real-time fluorescence quantitative polymerase chain reaction(qPCR),and compared among groups.Results On 1,3,5 and 7 d after modeling,the mNSS score of the model group was higher than that of the control group,the mNSS scores of the P2X4 agonist group and the NLRP1 agonist group were higher than that of the model group,while the mNSS score of the P2X4 antagonist group was lower than that of the model group,the differences were statistically significant(P<0.05).On 1,3,5 and 7 d after modeling,the levels of IL-1βand TNF-αin the model group were higher than those in the control group,the levels of IL-1βand TNF-αin the P2X4 agonist group and the NLRP1 agonist group were higher than those in the model group,while the levels of IL-1
作者
吴远水
黄小丽
徐建平
黄丹坪
熊成英
WU Yuanshui;HUANG Xiaoli;XU Jianping;HUANG Danping;XIONG Chengying(Department of Neurosurgery,ShangRao People's Hospital,Jiangxi Province,Shangrao 334000,China;School of Nursing,Jiangxi Medical College,Jiangxi Province,Shangrao 334000,China)
出处
《中国当代医药》
CAS
2024年第23期4-8,共5页
China Modern Medicine
基金
江西省卫生健康委科技计划项目(202410955)。
