八宝丹对脑缺血再灌注损伤大鼠自噬以及NLRP3炎症小体活化的影响  

Effects of Babao Dan on autophagy and activation of NLRP3 inflammasomes in rats with cerebral ischemia-reperfusion injury

作　　者：赵慎非 余文婷 黄贞伟 黄鸣清[2] 林思荣 ZHAO Shenfei;YU Wenting;HUANG Zhenwei;HUANG Mingqing;LIN Sirong(Fujian Drug Inspection Center,Fuzhou 350001,China;College of Pharmacy,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China;Fujian Institute for Food and Drug Quality Control,Fuzhou 350001,China)

机构地区：[1]福建省药品审核查验中心,福建福州350001 [2]福建中医药大学药学院,福建福州350122 [3]福建省食品药品质量检验研究院,福建福州350001

出　　处：《药学研究》2024年第8期729-733,共5页Journal of Pharmaceutical Research

基　　金：国家自然科学基金项目(No.81973437、82274080)。

摘　　要：目的研究八宝丹对大脑中动脉栓塞(middle cerebral artery occlusion,MCAO)大鼠脑缺血再灌注损伤的作用及可能机制。方法将健康雄性SD大鼠45只随机分为假手术组(Sham)、MCAO模型组以及八宝丹组,连续给药4 d后建立MCAO模型,24 h后采用Zea-Longa法进行神经功能评分;使用2,3,5-三苯基氯化四氮唑(TTC)染色法检测脑梗死体积;实时荧光定量聚合酶链式反应(qRT-PCR)法检测脑组织白介素-6(IL-6)、白介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、环氧化酶-2(COX-2)、一氧化氮合酶(iNOS)、单核细胞趋化蛋白-1(MCP-1)mRNA水平;蛋白质免疫印迹法(Western blotting)检测脑组织含NOD样受体热蛋白结构域相关蛋白3(NLRP3)、白介素-18(IL-18)、微管相关蛋白轻链3(LC3)、程序性死亡受体-1(Beclin1)、哺乳动物雷帕霉素靶蛋白(mTOR)蛋白表达水平。结果与模型组相比,八宝丹可减小MCAO大鼠脑梗死体积并降低神经功能评分(P<0.05),下调脑内IL-6、IL-1β、TNF-α、iNOS、COX-2、MCP-1等炎症介质的mRNA水平(P<0.05或P<0.01),降低脑组织中NLRP3、IL-18蛋白表达水平以及p-mTOR/mTOR比值,并上调Beclin1蛋白表达水平以及LC3-Ⅱ/LC3-Ⅰ比值(P<0.05、P<0.01、P<0.001)。结论八宝丹可能通过促进自噬和抑制NLRP3炎症小体活化而减少炎症介质释放,进而改善MCAO大鼠脑缺血再灌注损伤。Objective To study the effect of Babao Dan(BBD)on cerebral ischemia-reperfusion injury in middle cerebral artery occlusion(MCAO)rat and its possible mechanism.Methods 45 healthy male SD rats were randomly divided into Sham group,MCAO group and BBD group.MCAO model was established after four days of continuous administration.Zea-Longa method was used to score neurological deficits 24 hours later;2,3,5-Triphenyltetrazolium chloride(TTC)staining method to detect cerebral infarct volume in rats;quantitative real-time fluorescent polymerase chain reaction(qRT-PCR)was used to detect the mRNA level of interleukin-6(IL-6),interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),cyclooxygenase-2(COX-2),nitric oxide synthase(iNOS)and monocyte chemoattractant protein-1(MCP-1);The protein levels of NLR family Pyrin domain protein 3(NLRP3),interleukin-18(IL-18),microtubule-associated protein light chain 3(LC3),programmed death receptor-1(Beclin1)and mammalian target of rapamycin(mTOR)were detected by Western blotting.Results Compared with the MCAO group,BBD could significantly reduce the cerebral infarction volume and neurological function score of MCAO rats(P<0.05),and down-regulate mRNA levels of inflammatory mediators such as IL-6,IL-1β,TNF-α,iNOS,COX-2 and MCP-1(P<0.05,P<0.01).BBD can also down-regulate the protein expression of NLRP3,IL-18 and ratio of p-mTOR/mTOR,and up-regulate protein expression of Beclin1 and ratio of LC3-Ⅱ/LC3-I(P<0.05,P<0.01 and P<0.001).Conclusion BBD may reduce the release of inflammatory mediators by enhancing autophagy and inhibiting the activation of NLRP3 inflammasomes,thereby improving cerebral ischemia-reperfusion injury in MCAO rats.

关 键 词：八宝丹 脑缺血再灌注损伤 自噬 NOD样受体热蛋白结构域相关蛋白3炎症小体 

分 类 号：R285.5[医药卫生—中药学]