桃叶珊瑚苷通过调控Nrf2/NF-κB信号通路改善穿刺诱导的大鼠椎间盘退变  

Aucubin Ameliorates Puncture-Induced Intervertebral Disc Degeneration in Rats by Modulating the Nrf2/NF-κB Signaling Pathway

作　　者：王帅 贾欢欢 韦林 范小春 WANG Shuai;JIA Huan-Huan;WEI Lin;FAN Xiao-Chun(Dept.of Orthopedics,Nanchong Hospital of Traditional Chinese Medicine,Nanchong 637000 Sichuan,China)

机构地区：[1]南充市中医医院骨科,四川南充637000

出　　处：《广州中医药大学学报》2024年第12期3273-3282,共10页Journal of Guangzhou University of Traditional Chinese Medicine

基　　金：南充市科学技术局科研项目(编号:22YYJCYJ0064);南充市科学技术局科研项目(编号:23YYJCYJ0109);医学影像四川重点实验室科研资金项目(编号:MIKL202309)。

摘　　要：【目的】观察桃叶珊瑚苷对椎间盘退变大鼠的治疗作用及机制。【方法】将40只大鼠随机分为5组,即假手术组、模型组、桃叶珊瑚苷组、桃叶珊瑚苷+白藜芦醇[核转录因子kappaB(NF-κB)通路激活剂]组、桃叶珊瑚苷+ML385[Nrf2抑制剂]组,每组8只。除假手术组,其余各组大鼠采用纤维环穿刺法制备椎间盘退变模型。成功造模后,各组进行相应干预。干预结束后,采用免疫组织化学法检测椎间盘髓核组织中基质金属蛋白酶(MMP)-3和Ⅱ型胶原蛋白(COLⅡ)的表达,末端脱氧核苷酸转移酶dUTP缺口标记(TUNEL)测定椎间盘髓核细胞凋亡,比色法检测椎间盘髓核组织中丙二醛(MDA)、超氧化物歧化酶(SOD)、白细胞介素(IL)-1β、IL-6、肿瘤坏死因子α(TNF-α)的水平,Western Blot法测定椎间盘髓核组织中MMP-3、MMP-13、COLⅡ、Aggrecan、Bcl-2、caspase-3、p-p65、p65、磷酸化IκBα(p-IκBα)、IκBα、Nrf2、血红素加氧酶1(HO-1)、NAD(P)H:醌氧化还原酶1(NQO-1)蛋白表达水平。【结果】与假手术组比较,模型组大鼠椎间盘髓核组织中MMP-3阳性染色细胞比例升高、COLⅡ阳性染色细胞比例降低,髓核细胞凋亡比例升高,MDA水平升高、SOD水平降低,IL-1β、IL-6、TNF-α水平升高,MMP-3、MMP-13、caspase-3、p-p65/p65、p-IκBα/IκBα、细胞核p65蛋白表达水平均显著升高,而COLⅡ、Aggrecan、Bcl-2、细胞核Nrf2、HO-1、NQO-1蛋白表达水平均显著降低,差异均有统计学意义(P<0.05)。与模型组比较,桃叶珊瑚苷组椎间盘髓核组织MMP-3阳性染色细胞比例降低、COLⅡ阳性染色细胞比例升高,髓核细胞凋亡比例降低,MDA水平降低、SOD水平升高,IL-1β、IL-6、TNF-α水平降低,MMP-3、MMP-13、caspase-3、p-p65/p65、p-IκBα/IκBα、细胞核p65蛋白表达水平均显著降低,而COLⅡ、Aggrecan、Bcl-2、细胞核Nrf2、HO-1、NQO-1蛋白表达水平均显著升高(P<0.05)。白藜芦醇和ML385能够在一定程度上逆转桃�Objective To observe the therapeutic effect and mechanism of Aucubin on rats with intervertebral disc degeneration(IDD).Methods Forty rats were randomly divided into five groups,i.e.,sham-operated group,model group,Aucubin group,Aucubin+Resveratrol[nuclear factor-kappa B(NF-κB)pathway activator]group,and Aucubin+ML385(Nrf2 inhibitor)group,with eight rats in each group.Except for the sham-operated group,rats in all groups were prepared for the IDD model using the annulus fibrosus puncture method.After successful modeling,each group was given corresponding intervention.After the end of intervention,the expression of matrix metalloproteinase(MMP)-3 and collagen typeⅡ(COLⅡ)in the nucleus pulposus of intervertebral discs was detected by immunohistochemical analysis.Terminal deoxynucleotidyl transferase dUTP nick labeling(TUNEL)was used to determine the apoptosis of nucleus pulposus cells in the intervertebral discs.And malondialdehyde(MDA)and superoxide dismutase(SOD),interleukin(IL)-1β,IL-6,and tumor necrosis factorα(TNF-α)levels in the nucleus pulposus of intervertebral discs were detected by colorimetry.Western Blot was used to determine the protein expression levels of MMP-3,MMP-13,COLⅡ,Aggrecan,Bcl-2,caspase-3,p-p65,p65,phosphorylated IκBα(p-IκBα),IκBα,Nrf2,heme oxygenase 1(HO-1),and NAD(P)H:nadph quinine oxidoreductase-1(NQO-1).Results Compared with the sham-operated group,the proportion of MMP-3-positively stained cells was increased in rats in the model group,COLⅡ-positively stained cells was decreased,the apoptosis rate of nucleus pulposus cells was increased,MDA level was increased,SOD level was decreased,the levels of IL-1β,IL-6 and TNF-αwere elevated,and the protein expression levels of MMP-3,MMP-13,caspase-3,p-p65/p65,p-IκBα/IκBαand nuclear p65 were decreased,protein expression levels of COLⅡ,Aggrecan,Bcl-2,nuclear Nrf2,HO-1,NQO-1 were increased in the nucleus pulposus of intervertebral discs,the differences being significantly statistical(P<0.05).Compared with the model group,th

关 键 词：桃叶珊瑚苷 椎间盘退变 髓核 氧化应激 炎症 大鼠 

分 类 号：R285.5[医药卫生—中药学]