灵仙新苷对脑缺血再灌注损伤大鼠脑组织氧化应激和炎症反应的调控及神经细胞凋亡的影响  

Effects of Clematichinenoside AR on oxidative stress and inflammatory response regulation and nerve cell apoptosis in brain tissue of rats with cerebral ischemia reperfusion injury

作　　者：熊建 屈战利 任瑜 季一飞 XIONG Jian;QU Zhanli;REN Yu;JI Yifei(Department of Neurology,Nanchong Central Hospital(The Second Clinical Medical College of North Sichuan Medical College),Nanchong,Sichuan 637000,China)

机构地区：[1]南充市中心医院·川北医学院第二临床医学院神经内科,四川南充637000

出　　处：《湖南中医药大学学报》2024年第4期551-556,共6页Journal of Hunan University of Chinese Medicine

基　　金：四川省自然科学基金项目(2022NSFSC0756)。

摘　　要：目的研究灵仙新苷对脑缺血再灌注损伤大鼠脑组织氧化应激反应、神经细胞凋亡以及炎症因子的作用。方法45只大鼠分成对照组、模型组(脑缺血再灌注损伤大鼠)、灵仙新苷低剂量组(8 mg/kg)、灵仙新苷中剂量组(16 mg/kg)、灵仙新苷高剂量组(32 mg/kg),每组9只。治疗5 d后,对大鼠进行神经功能评分,TTC染色法检测脑梗死体积,TUNEL法检测细胞凋亡,Western blot检测脑组织中切割型胱天蛋白酶-3(cleaved-cysteine aspartic acid specific protease-3,C-Caspase-3)、切割型胱天蛋白酶-9(cleaved-cysteine aspartic acid specific protease-9,C-Caspase-9)、核因子κB p65(nuclear factor-κB p65,NF-κB p65)蛋白水平,二硝基苯肼法检测乳酸脱氢酶(lactate dehydrogenase,LDH)水平,硫代巴比妥酸法检测丙二醛(malondialdehyde,MDA)水平,黄嘌呤氧化法检测超氧化物歧化酶(superoxide dismutase,SOD)水平,比色法检测谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)水平,ELISA法检测白细胞介素-1β(interleukin-1β,IL-1β)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)含量。结果与对照组比较,模型组大鼠神经功能评分升高(P<0.05),脑梗死体积增加(P<0.05),细胞凋亡指数和C-Caspase-3、C-Caspase-9蛋白水平升高(P<0.05),LDH、MDA水平升高(P<0.05),SOD、GSH-Px水平降低(P<0.05),IL-1β、TNF-α含量升高(P<0.05),NF-κB p65蛋白表达水平升高(P<0.05)。与模型组比较,灵仙新苷低、中、高剂量组大鼠神经功能评分依次降低(P<0.05),脑梗死体积依次减少(P<0.05),细胞凋亡指数和C-Caspase-3、C-Caspase-9蛋白水平依次降低(P<0.05),LDH、MDA水平依次降低(P<0.05),SOD、GSH-Px水平依次升高(P<0.05),IL-1β、TNF-α含量依次降低(P<0.05),NF-κB p65蛋白表达水平依次降低(P<0.05)。结论灵仙新苷能降低脑缺血再灌注损伤大鼠脑组织氧化应激反应水平,减少细胞凋亡因子C-Caspase-3、C-Caspase-9的表达,抑制炎症介质释放,其机制可能与下调NF-κObjective To study the effects of Clematichinenoside AR on oxidative stress response,nerve cell apoptosis,and inflammatory factors in the brain tissue of rats with cerebral ischemia reperfusion injury.Methods Forty-five rats were divided into control group,model group(rats with cerebral ischemia reperfusion injury),and Clematichinenoside AR low-(8 mg/kg),medium-(16 mg/kg),and high-(32 mg/kg)dose groups,with nine rats in each group.After 5 d of treatment,the rats were scored for neurological function.TTC staining was used to check cerebral infarction volume,TUNEL method to measure the degree of apoptosis,Western blot to determine the protein levels of cleaved-cysteine aspartic acid specific protease-3(C-Caspase-3),cleaved-cysteine aspartic acid specific protease-9(C-Caspase-9),and nuclear factor-κB p65(NF-κB p65)in the brain tissue,dinitrophenylhydrazine method to examine the lactate dehydrogenase(LDH)level,thiobarbituric acid method to measure the malondialdehyde(MDA)level,xanthine oxidation method to check the superoxide dismutase(SOD)level,colorimetry to examine the glutathione peroxidase(GSH-Px)level,and ELISA to determine the content of interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α).Results Compared with the control group,the neurological function score in the model group increased(P<0.05),the cerebral infarction volume increased(P<0.05),the apoptosis index and the protein levels of C-Caspase-3 and C-Caspase-9 increased(P<0.05),the levels of LDH and MDA increased(P<0.05),SOD and GSH-Px levels decreased(P<0.05),the content of IL-1βand TNF-αincreased(P<0.05),and the protein expression level of NF-κB p65 increased(P<0.05).Compared with the model group,the neurological function score in Clematichinenoside AR low-,medium-,and high-dose groups decreased successively(P<0.05),the cerebral infarction volume decreased successively(P<0.05),the apoptosis index and the protein levels of C-Caspase-3 and C-Caspase-9 decreased successively(P<0.05),the levels of LDH and MDA decreased successively(P<0.05),the

关 键 词：灵仙新苷 脑缺血再灌注损伤 氧化应激 细胞凋亡 炎症因子 

分 类 号：R285.5[医药卫生—中药学]