摘要
目的:探究丹皮酚(Pae)通过上调微小RNA-339-5p(miR-339-5p)靶向高迁移率族蛋白B1(HMGB1)对神经病理性疼痛大鼠炎症反应的影响。方法:通过L5神经结扎横切构建神经病理性疼痛大鼠模型,将72只SD大鼠随机分为6组,假手术组(Sham)、模型组(Model)、Pae组、Pae+antagomir-NC组、Pae+miR-339-5p-antagomir组、Pae+miR-339-5p-antagomir+HMGB1抑制剂-甘草酸(GA)组,每组12只,给药7 d。手术前1 d及手术后3 d、11 d,测定大鼠行为学机械痛阈值(MWT)、热缩足反射潜伏期(TWL),手术后第12天,酶联免疫吸附法测定脊髓组织中肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)水平,实时荧光定量聚合酶链式反应(RT-PCR)法测定脊髓组织中miR-339-5p、HMGB1、IL-1β、TNF-αmRNA的表达,蛋白免疫印迹法(Western Blot)检测脊髓组织中HMGB1、IL-1β、TNF-α蛋白表达,双荧光素酶报告实验分析miR-339-5p与HMGB1的靶向关系。结果:术前1 d,各组大鼠MWT、TWL比较差异无统计学意义(P>0.05)。与Sham组比较,Model组大鼠术后MWT、TWL及miR-339-5p水平降低,IL-1β、TNF-α水平,HMGB1、IL-1β、TNF-αmRNA及蛋白水平升高(P<0.05);与Model组比较,Pae组大鼠术后MWT、TWL、miR-339-5p水平升高,IL-1β、TNF-α水平及HMGB1、IL-1β、TNF-αmRNA及蛋白水平降低(P<0.05);与Pae组比较,Pae+antagomir-NC组大鼠各指标差异无统计学意义(P>0.05),Pae+miR-339-5p-antagomir组大鼠术后MWT、TWL及miR-339-5p水平降低,IL-1β、TNF-α水平,HMGB1、IL-1β、TNF-αmRNA及蛋白水平升高(P<0.05);与Pae+miR-339-5p-antagomir组比较,Pae+miR-339-5p-antagomir+GA组大鼠术后MWT、TWL升高,IL-1β、TNF-α水平,HMGB1、IL-1β、TNF-αmRNA及蛋白水平降低(P<0.05);mir-339-5p与HMGB1存在靶向关系。结论:Pae通过上调miR-339-5p,靶向抑制HMGB1的表达,减轻神经病理性疼痛大鼠的炎症反应。
Objective:To explore the effect of paeonol(Pae)on the inflammatory response in rats with neuropathic pain by up-regulating microRNA-339-5p(miR-339-5p)and targeting high mobility group box B1(HMGB1).Methods:The rat model of neuropathic pain was established by ligation and transection of the L5 nerve.A total of 72 Sprague-Dawley(SD)rats were randomly divided into 6 groups,including the sham group,model group,Pae group,Pae+antagomir-NC group,Pae+miR-339-NC group,Pae+miR-339-5p-antagomir group,and Pae+miR-339-5p-antagomir+HMGB1 inhibitor-glycyrrhizic acid(GA)group,with 12 rats in each group and administered for 7 days.On 1 day before the operation,3 days,and 11 days after the operation,the mechanical withdrawal latency(MWL)and thermal withdrawal latency(TWL)of rats were measured.At the 12th day after the operation,enzyme-linked immunosorbent assay(ELISA)was used to detect tumor necrosis in spinal cord tissue,factor-α(TNF-α),and interleukin-1β(IL-1β)levels;real-time fluorescent quantitative PCR(RT-PCR)method was used to detect miR-339-5p,HMGB1,IL-1β,TNF-αin spinal cord tissue mRNA expression;Western Blot was used to detect HMGB1,IL-1β,TNF-αprotein expression in spinal cord tissue;dual luciferase reporter assay was used to analyze the targeting relationship between miR-339-5p and HMGB1.Results:At 1 day before the operation,there was no significant difference in MWT and TWL among the rats in each group(P>0.05).Compared with the sham group,the levels of MWT,TWL,and miR-339-5p decreased;the levels of IL-1βand TNF-α,HMGB1,IL-1β,TNF-αmRNA,and protein increased in the model group after the operation(P<0.05).Compared with the Model group,the levels of MWT,TWL,and miR-339-5p increased;the levels of IL-1β,TNF-α,HMGB1,IL-1β,TNF-αmRNA,and protein decreased in the Pae group after the operation(P<0.05).Compared with the Pae group,there was no significant difference in the indicators of the rats in the Pae+antagomir-NC group(P>0.05);the postoperative levels of MWT,TWL,and miR-339-5p decreased;the levels of IL-1β,TNF
作者
王振
王玉萍
张爱忠
WANG Zhen;WANG Yuping;ZHANG Aizhong(PKUCare Luzhong Hospital,Zibo 255400,Shandong,China)
出处
《中西医结合心脑血管病杂志》
2023年第14期2600-2606,共7页
Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease
