甜叶悬钩子苷对脊髓损伤小鼠运动功能障碍和神经炎症的改善作用及其机制  

Improvement effect of rubusoside on motor dysfunction and neuroinflammation in mice with spinal cord injury and its mechanism

作　　者：杨爽 许娜[3] 张剑旭 孙成彪 王燕 董明鑫 刘文森 YANG Shuang;XU Na;ZHANG Jianxu;SUN Chengbiao;WANG Yan;DONG Mingxin;LIU Wensen(Military Veterinary Research Institute,Academy of Military Medical Sciences,Academy of Military Sciences,Changchun 130117,China;Department of Physiology and Pathophysiology,School of Medical Sciences,Yanbian University,Yanji 133002,China;Jilin Medical College,Jilin 132000,China)

机构地区：[1]军事科学院军事医学研究院军事兽医研究所,吉林长春130117 [2]延边大学医学院生理学与病理生理学教研室,吉林延吉133002 [3]吉林医药学院,吉林吉林132000

出　　处：《吉林大学学报（医学版）》2024年第2期326-335,共10页Journal of Jilin University：Medicine Edition

基　　金：吉林省教育厅科学研究项目(JJKH20210503KJ)。

摘　　要：目的:探讨甜叶悬钩子苷(RUB)对小鼠脊髓损伤(SCI)和神经炎症的影响,并阐明其作用机制。方法:将48只雌性昆明小鼠随机分为假手术组、SCI组、SCI+低剂量RUB组和SCI+高剂量RUB组,每组12只;采用脊髓损伤行为学(BBB)评分法评估SCI小鼠后肢运动功能,脊髓组织含水量法检测SCI小鼠脊髓水肿情况,实时荧光定量PCR(RT-qPCR)法检测各组小鼠促炎细胞因子环氧化酶2(COX-2)、白细胞介素1β(IL-1β)和肿瘤坏死因子α(TNF-α)mRNA表达水平,ELISA法检测各组小鼠血清中炎症因子水平,HE染色观察各组小鼠脊髓组织病理形态学,免疫荧光法检测各组小鼠脊髓组织中小胶质细胞活化情况,Western blotting法检测SCI小鼠脊髓组织中相关蛋白表达水平。结果:BBB评分,与假手术组比较,SCI组小鼠评分低至0分;与SCI组比较,SCI+低剂量RUB组和SCI+高剂量RUB组小鼠BBB评分逐步升高。脊髓组织含水量法检测,与假手术组比较,SCI组小鼠脊髓组织含水量明显升高(P<0.01);与SCI组比较,SCI+低剂量RUB组和SCI+高剂量RUB组小鼠脊髓组织含水量明显降低(P<0.01)。RT-qPCR法检测,与假手术组比较,SCI组小鼠脊髓组织中COX-2、IL-1β和TNF-αmRNA表达水平明显升高(P<0.001);与SCI组比较,SCI+低剂量RUB组和SCI+高剂量RUB组小鼠脊髓组织中COX-2、IL-1β和TNF-αmRNA表达水平明显降低(P<0.001)。ELISA法检测,与假手术组比较,SCI组小鼠血清中IL-1β(P<0.01)和TNF-α(P<0.001)水平升高;与SCI组比较,SCI+低剂量RUB组和SCI+高剂量RUB组小鼠血清中IL-1β和TNF-α水平降低(P<0.001);Western blotting法检测,与假手术组比较,SCI组小鼠脊髓组织中核因子κB(NF-κB)抑制因子α(IκB-α)、磷酸化IκB-α(p-IκB-α)、磷酸化p65(p-p65)、p-65、磷酸化p38(p-p38)、磷酸化细胞外调节蛋白激酶(p-ERK)和磷酸化c-Jun氨基末端激酶(p-JNK)蛋白表达水平明显升高(P<0.05或P<0.001);与SCI组比较,SCI+低剂量RUB组和SCI+高剂量RUB组小鼠脊�Objective:To discuss the effect of rubusoside(RUB)on the spinal cord injury(SCI)and neuroinflammation in the mice,and to clarify its mechanism.Methods:A total of 48 female Kunming mice were randomly divided into sham operation group,SCI group,SCI+low dose of RUB group,and SCI+high dose of RUB group,and there were 12 mice in each group.Spinal cord injury behavior(BBB)scoring method was used to assess the motor function of the hind limbs of the SCI mice;water content method was used to detect the spinal cord edema of the SCI mice;real-time fluorscence quantitative PCR(RT-qPCR)method was used to detect the expression levels of pro-inflammatory cytokines cyclooxygenase-2(COX-2),interleukin-1β(IL-1β),and tumor necrosis factor-α(TNF-α)mRNA of the mice in various groups;ELISA method was used to detect the levels of inflammatory factors in serum of the mice in various groups;HE staining was used to observe the pathomorphology of spinal cord tissue of the mice in various groups;immunofluorescence was used to detect the activation of microglia in spinal cord tissue of the mice in various groups;Western blotting method was used to detect the expression levels of related proteins in spinal cord tissue of the mice in various groups.Results:The BBB score analysis results showed that compared with sham operation group,the score of the mice in SCI group was decreased to 0;compared with SCI group,the BBB scores of the mice in SCI+low dose of RUB group and SCI+high dose of RUB group were gradually increased.The spinal cord tissue water content detection results showed that compared with sham operation group,the water content in spinal cord tissue of the mice in SCI group was significantly increased(P<0.01);compared with SCI group,the water contents in spinal cord tissue of the mice in SCI+low dose of RUB group and SCI+high dose of RUB group were significantly decreased(P<0.01).The RT-qPCR results showed that compared with sham operation group,the expression levels of COX-2,IL-1β,and TNF-αmRNA in spinal cord tissue of the mic

关 键 词：甜叶悬钩子苷 脊髓损伤 神经炎症 核因子ΚB 丝裂原活化蛋白激酶 

分 类 号：R363.1[医药卫生—病理学]