黄芪总黄酮通过调节Notch-Wnt信号通路对强直性脊柱炎模型大鼠炎症反应和成纤维细胞成骨转化的影响  

Effects of total flavonoids of astragalus on inflammatory response and osteogenic transformation of fibroblasts in ankylosing spondylitis model rats by regulating Notch-Wnt signaling pathway

作　　者：焦士军 李岩 韦中阳[1] 刘瑞端[2] JIAO Shijun;LI Yan;WEI Zhongyang;LIU Ruiduan(Department of Orthopedic Surgery,Anyang People's Hospital,Anyang 455000,China;Department of Spine Surgery,the Affiliated Hospital of Guilin Medical University,Guilin 541001,China)

机构地区：[1]安阳市人民医院骨外一科,河南安阳455000 [2]桂林医学院附属医院脊柱外科,广西桂林541001

出　　处：《广东药科大学学报》2022年第3期43-48,共6页Journal of Guangdong Pharmaceutical University

基　　金：广西医疗卫生适宜技术开发与推广应用项目(S2019063)。

摘　　要：目的 探究黄芪总黄酮对强直性脊柱炎(AS)模型大鼠炎症反应和成纤维细胞成骨转化的影响及作用机制。方法 将72只大鼠随机分为6组,每组12只,分别为空白对照组、模型组、黄芪总黄酮低、中、高剂量组(25、50、100 mg/kg)和阳性药物组(柳氮磺胺吡啶825 mg/kg)。除空白对照组外,其余各组大鼠建立AS大鼠模型,药物干预28 d后,ELISA分别检测血清和脊柱组织中炎症因子(IL-6、IL-1β、IL-8),显色法检测成纤维细胞成骨型表达标志物[碱性磷酸酶(ALP)活性、骨钙素(BGP)、Ⅰ型前胶原氨基端前肽(PINP)含量]的变化,Western blot检测成骨分化相关细胞因子[骨形态发生蛋白2(BMP-2)、转化生长因子-β1(TGF-β1)]和Notch-Wnt信号通路相关蛋白表达。结果 与空白对照组比较,模型组大鼠血清、脊柱组织中IL-6、IL-1β和IL-8水平升高(P<0.05),血清ALP活性、BGP含量升高(P<0.05),PINP含量降低(P<0.05),脊柱组织中BMP-2、TGF-β1蛋白表达水平降低(P<0.05),Notch1、Hes1、Wnt1、β-catenin蛋白表达水平均升高(P<0.05);与模型组比较,黄芪总黄酮中、高剂量组和阳性药物组血清、脊柱组织中IL-6、IL-1β和IL-8水平降低(P<0.05),血清ALP活性、BGP含量降低(P<0.05),PINP含量升高(P<0.05),脊柱组织中BMP-2、TGF-β1蛋白表达水平升高(P<0.05),Notch1、Hes1、Wnt1、β-catenin蛋白表达水平均降低(P<0.05)。结论 黄芪总黄酮可减轻AS模型大鼠炎症反应,改善成纤维细胞成骨转化,可能与抑制Notch-Wnt信号通路相关。Objective To explore the effects and mechanism of total flavonoids of astragalus(TFA) on inflammatory response and osteogenic transformation of fibroblasts in ankylosing spondylitis(AS) model rats.Methods 72 rats were randomly divided into 6 groups with 12 rats in each group, namely the blank control group,the model group, low, middle and high doses of TFA groups(25, 50, 100 mg/kg) and positive drug group(825 mg/kg of sulfasalazine). Except the blank control group, AS model rats were established in other groups. After 28 days of drug intervention, the inflammatory factors(IL-6, IL-1β, IL-8) in serum and spinal tissue were detected by ELISA. The changes of osteoblast expression markers such as alkaline phosphatase(ALP) activity, bone gla protein(BGP) and procollagen type I N-terminal propeptide(PINP) were detected by chromogenic method. The expressions of osteogenic differentiation-related cytokines such as bone morphogenetic protein 2(BMP-2),transforming growth factor-β1(TGF-β1) and Notch-Wnt signaling pathways-related proteins were measured by western blot. Results Compared with the blank control group, the levels of IL-6, IL-1β and IL-8 in serum and spinal tissues of model group were increased, and serum ALP activity and BGP content were enhanced, but the content of PINP was reduced(P<0.05). At the same time, the protein levels of BMP-2 and TGF-β1 in the spinal tissues were decreased, while the expressions of Notch1, Hes1, Wnt1 and β-catenin were increased(P<0.05).Compared with the model group, the levels of IL-6, IL-1β and IL-8 in serum and spinal tissues were declined in middle and high doses of TFA groups and positive drug group(P<0.05). Serum ALP activity and BGP content were lowered, but the content of PINP was enhanced. The levels of BMP-2 and TGF-β1 in spinal tissues were elevated, but the levels of Notch1, Hes1, Wnt1 and β-catenin were declined(P<0.05). Conclusion Total flavonoids of astragalus can relieve the inflammatory response and improve the osteogenic transformation of fibroblasts in AS

关 键 词：黄芪总黄酮 强直性脊柱炎 炎症反应 成纤维细胞成骨转化 Notch-Wnt信号通路 

分 类 号：R285.5[医药卫生—中药学]