苦参总黄酮对大鼠局灶性脑缺血再灌注损伤的保护作用及相关机制  

Protective Effects and Related Mechanisms of Total Flavonoids of Sophora flavescens on Focal Cerebral Ischemia-reperfusion Injury in Rats

作　　者：许诺 魏玮 吴勤研 XU Nuo;WEI Wei;WU Qinyan(Qinhuai Medical District,Eastern Theater of PLA Theater Command,Nanjing 210002,China;General Hospital of Eastern Theater Command,Nanjing 210016,China)

机构地区：[1]东部战区总医院秦淮医疗区,江苏南京210002 [2]东部战区总医院,江苏南京210016

出　　处：《北华大学学报（自然科学版）》2023年第6期759-765,共7页Journal of Beihua University（Natural Science）

基　　金：江苏省科教强卫工程基金项目(YXZXB2016006);东部战区总医院院内基金项目(2015017)。

摘　　要：目的 探讨苦参总黄酮对大鼠局灶性脑缺血再灌注损伤的保护作用及相关机制.方法 应用大脑中动脉栓塞法构建大鼠局灶性脑缺血再灌注模型,将造模成功的40只大鼠随机分为模型组、50 mg/kg苦参总黄酮组、100 mg/kg苦参总黄酮组、200 mg/kg苦参总黄酮组;另取10只大鼠作为对照组(手术步骤同模型组,但不插入尼龙线阻断血流).对照组和模型组分别给予生理盐水灌胃,苦参总黄酮组造模后给予相对应剂量的苦参总黄酮灌胃,7 d后取材.分别于缺血再灌注1、3、7 d使用改良的Longa评分系统进行神经功能评分.应用酶联免疫吸附法测定血清白细胞介素-1(IL-1)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)浓度.应用RT-PCR和Western blot检测大鼠脑组织中无翅型MMTV整合位点家族成员3a(Wnt3a)、丝氨酸/苏氨酸激酶(GSK-3β)、β-连环蛋白肽(β-catenin)、G1/S-特异性周期蛋白-D1(CyclinD1)mRNA和蛋白表达.结果 缺血再灌注时间1、3、7 d时苦参总黄酮不同剂量组大鼠神经功能缺损评分均明显低于模型组,随着苦参总黄酮剂量的增加大鼠神经功能缺损评分明显降低(P<0.05);随着缺血再灌注时间的增加大鼠神经功能缺损评分逐渐升高,苦参总黄酮组大鼠神经功能缺损评分逐渐降低(P<0.05).模型组大鼠血清IL-1、IL-6、TNF-α含量明显高于对照组(P<0.01);苦参总黄酮各剂量组大鼠血清IL-1、IL-6、TNF-α含量均明显低于模型组,随着苦参总黄酮剂量的增加大鼠血清IL-1、IL-6、TNF-α含量逐渐降低(P<0.05).模型组大鼠Wnt3a、β-catenin、CyclinD1 mRNA和蛋白相对表达量明显高于对照组,GSK-3β mRNA和蛋白相对表达量明显低于对照组(P<0.01);不同剂量的苦参总黄酮组大鼠Wnt3a、β-catenin、CyclinD1 mRNA和蛋白相对表达量均明显低于模型组,GSK-3β mRNA和蛋白相对表达量明显高于模型组(P<0.05);随着苦参总黄酮剂量的增加,大鼠Wnt3a、β-catenin、CyclinD1Objective To explore the protective effect and related mechanisms of total flavonoids of Sophora flavescens on focal cerebral ischemia-reperfusion injury in rats.Method A rat model of focal cerebral ischemia-reperfusion was established by using the middle cerebral artery embolization method.The successfully established forty rats were randomly divided into model group,50 mg/kg total flavonoid group,100 mg/kg total flavonoid group and 200 mg/kg total flavonoid group;Another 10 rats were selected as the control group,with the same surgical procedures as the model group,but without inserting nylon thread to block blood flow.The control group and model group were given physiological saline by gavage,while the total flavonoid group of Sophora flavescens was given corresponding doses of total flavonoid of Sophora flavescens by gavage after modeling.Samples were taken 7 days later.The improved Longa scoring system was used to assess neurological function on day 1,day 3 and day 7 of ischemia-reperfusion.After taking blood from the abdominal aorta,the rats were euthanized.The serum interleukin-1(IL-1),Interleukin 6(IL-6) and tumor necrosis factor by enzyme-linked immunosorbent assay-α(TNF-α) concentration were determined.RT-PCR and Western blot were used to detect wingless MMTV integration site family member 3a(Wnt3a),serine/Threonine kinase(GSK-3β) in rat brain tissue,β-catenin peptide(β-catenin),G1/S-specific cyclinD1(CyclinD1) mRNA and protein expression.Results The time of ischemia-reperfusion for 1 day,3 days and 7 days,the neurological deficit scores of rats in the different doses of total flavonoid of Sophora flavescens group were significantly lower than those in the model group.With the increase of total flavonoid dose of Sophora flavescens,the neurological deficit scores of rats significantly decreased(P<0.05);As the ischemia-reperfusion time increases,the neurological deficit score of rats gradually increases,while the total flavonoid group in the Sophora flavescens group gradually decreases.The brain wate

关 键 词：缺血性脑卒中 再灌注损伤 苦参总黄酮 神经功能评分 脑梗死体积 炎症反应 作用机制 

分 类 号：R285.5[医药卫生—中药学]