文献类型：期刊文献

中文题名：藤黄健骨胶囊对膝骨关节炎鼠软骨组织Notch1、Jagged1、炎性反应因子及基质金属蛋白酶表达的影响

英文题名：Effects of tenghuangjiangu capsule on the expression of Notch1,Jagged1,inflammatory factors,and matrix metalloproteinases in rats cartilage tissue of knee osteoarthritis model

作者：蔡成成[1];颜春鲁[2];安方玉[3];孙柏[2];王霞霞[2];邓婕[4];孟祥睿[4];张金龙[4];石瑶[5];袁灵青[5];席永斌[1]

第一作者：蔡成成

机构：[1]兰州市第二人民医院骨科,兰州730046;[2]甘肃中医药大学中西医结合学院.中医外科教研室,兰州730000;[3]甘肃中医药大学教学实验实训中心生物化学实验室,兰州730000;[4]甘肃中医药大学第一临床医学院内科教研室,兰州730000;[5]甘肃中医药大学基础医学院微生物与免疫学教研室,兰州730000

第一机构：兰州市第二人民医院骨科,兰州730046

年份：2023

卷号：16

期号：1

起止页码：41

中文期刊名：中华骨质疏松和骨矿盐疾病杂志

外文期刊名：Chinese Journal Of Osteoporosis And Bone Mineral Research

收录：CSTPCD;;北大核心:【北大核心2020】；CSCD:【CSCD2023_2024】；

基金：甘肃省中医药管理局科研项目(GZKP-2021-34);甘肃省教育科技创新项目(2022QB-091);兰州市卫生健康科技发展项目(2021004);甘肃中医药大学科学研究与创新基金项目(2021KCYB-12)。

语种：中文

中文关键词：藤黄健骨胶囊;膝骨关节炎;Notch/Jagged1信号通路;炎性反应因子;基质金属蛋白酶

外文关键词：Tenghuangjiangu capsule;knee osteoarthritis;Notch/Jagged1 signal pathway;inflammatory cytokines;matrix metalloproteinases

摘要：目的探讨Notch/Jagged1信号通路对膝骨关节炎(knee osteoarthritis,KOA)模型鼠软骨组织炎性反应因子及基质金属蛋白酶(matrix metalloproteinase,MMPs)表达的影响,并揭示藤黄健骨胶囊可能的干预机制。方法60只SD雌性大鼠随机分为假手术组、模型组、仙灵骨葆胶囊组[0.27 g/(kg·d)]及藤黄健骨胶囊干预组[高剂量0.36 g/(kg·d)、中剂量0.18 g/(kg·d)、低剂量0.09 g/(kg·d)],每组10只。除假手术组外,其余5组采用改良Hulth法构建KOA大鼠模型,模型制备成功后,分别给予仙灵骨葆胶囊及相应剂量的藤黄健骨胶囊灌胃给药,假手术组和模型组给予0.9%(质量分数)氯化钠注射液。给药2个月后股动脉采血处死大鼠,取患侧膝关节软骨采用苏木精-伊红(HE)染色观察关节软骨组织的形态变化;ELISA法检测关节软骨组织中白细胞介素-1β(interleukin-1β,IL-1β)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、Notch1和Jagged1含量变化;免疫组化法观察关节软骨组织中Col-Ⅱ、MMP-9、MMP-13、Notch1和Jagged1蛋白表达变化。结果与假手术组比较,模型组关节面脱落、严重破坏,软骨细胞分布紊乱;模型组软骨细胞Mankin(5.56±0.85)评分、IL-1β(11.13±2.19)、TNF-α(98.29±9.13)、Notch1(5.03±0.37)和Jagged1(222.54±36.87)含量均显著升高,MMP-9(1.54±0.72)、MMP-13(0.41±0.04)、Notch1(0.39±0.05)和Jagged1(0.38±0.05)等蛋白的免疫反应性均显著增强,Col-Ⅱ(0.15±0.04)蛋白的免疫反应性则显著降低(P<0.05)。与模型组比较,藤黄健骨胶囊0.36 g/(kg·d)剂量组软骨细胞形态和分布趋于正常;其软骨组织Mankin(2.86±0.48)评分显著降低,Col-Ⅱ(0.39±0.03)蛋白的免疫反应性则显著增强(P<0.05);藤黄健骨胶囊0.36、0.18 g/(kg·d)剂量组Jagged1(170.54±41.09、183.46±33.12)含量显著降低,MMP-9(0.68±0.25、1.14±0.49)和Jagged1(0.21±0.05、0.26±0.07)等蛋白的免疫反应性也均显著降低(P<0.05);藤黄健骨胶囊0.36、0.18、0.09 g/(kg·d)剂量组IL-1β(5.71±1.03、8.71±1.53、8.81±2.31)、TNF-α(63.42±10.01、67.46±11.68、71.88±15.63)和Notch1(2.95±0.39、3.56±0.37、4.22±0.33)含量均显著降低,同时MMP-13(0.21±0.02、0.27±0.04、0.31±0.09)和Notch1(0.19±0.06、0.24±0.05、0.28±0.04)等蛋白的免疫反应性也均显著降低。结论藤黄健骨胶囊可能通过促进KOA模型鼠关节软骨组织Notch/Jagged1信号通路下游蛋白Col-Ⅱ表达,抑制其Notch/Jagged1信号通路关键蛋白Notch、Jagged1、炎性反应因子及MMPs表达,延缓关节软骨退变,从而改善KOA的临床症状。
Objective To explore the roles of Notch/Jagged1 signaling pathway in inflammatory factors and matrix metalloproteinases(MMPs)of cartilage tissue in rat model of knee osteoarthrosis,and to evaluate the possible mechanism of tenghuangjiangu capsule in treating knee osteoarthritis(KOA).Methods Sixty SD female rats were randomly divided into sham operation group,model group,xianlinggubao capsule group[(0.27 g/(kg·d)],and tenghuangjiangu capsule intervention groups[high-dose 0.36 g/(kg·d),medium-dose 0.18 g/(kg·d),and low-dose 0.09 g/(kg·d)],and there were 10 rats in each group.Except for the sham operation group,KOA model rats were induced by modified Hulth method in the other five groups.After successful modeling,xianlinggubao capsule and the corresponding dose of tenghuangjiangu capsule was given oraly,the sham operation group and model group were given normal saline.On the 2nd month after administration,rats were sacrificed by femoral artery blood collection.HE staining was adopted to observe the morphologic changes of cartilage articularis of affected side.ELISA measured the contents of interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),Notch1,and Jagged1.The expression of Col-Ⅱ,MMP-9,MMP-13,Notch1,and Jagged1 observed by immunohistochemistry staining.Results Compared with the sham operation group,the morphologic changes of cartilage articularis in the model group showed abscission,seriously damaged,disorder distribution.The Mankin(5.56±0.85)score and the concentrations of IL-1β(11.13±2.19),TNF-α(98.29±9.13),Notch1(5.03±0.37),and Jagged1(222.54±36.87)were significantly increased.The level of MMP-9(1.54±0.72),MMP-13(0.41±0.04),Notch1(0.39±0.05),and Jagged1(0.38±0.05)significantly increased;however,the level of ColⅡ(0.15±0.04)significantly decreased(P<0.05).Compared with the model group,cartilage structure and distribution was tended to be normal in the 0.36 g/(kg·d)group of tenghuang jiangu capsule.The Mankin(2.86±0.48)score in the 0.36 g/kg group of tenghuangjiangu capsule were significantly lower than that in the model group,yet the level of ColⅡ(0.39±0.03)significantly increased(P<0.05).The concentration of Jagged1(170.54±41.09,183.46±33.12)and the level of MMP-9(0.68±0.25,1.14±0.49),Jagged1(0.21±0.05,0.26±0.07)in the 0.36,0.18 g/(kg·d)groups of tenghuangjiangu capsule were significantly decreased than those of in the model group(P<0.05).The concentration of IL-1β(5.71±1.03,8.71±1.53,8.81±2.31),TNF-α(63.42±10.01,67.46±11.68,71.88±15.63),and Notch1(2.95±0.39,3.56±0.37,4.22±0.33)were evidently decreased in the 0.36,0.18,0.09 g/(kg·d)groups of tenghuangjiangu capsule,and meanwhile the level of MMP-13(0.21±0.02,0.27±0.04,0.31±0.09),and Notch1(0.19±0.06,0.24±0.05,0.28±0.04)were also decreased than those of in the model group(P<0.05).Conclusions Intervention of tenghuangjiangu capsule maybe promote the expression of Col-Ⅱin dowstream protein of the Notch/Jagged1 signaling pathway,inhibiting the expression of Notch,Jagged1,inflammatory factors,and MMPs of the Notch/Jagged1 signaling pathway in the articular cartilage of KOA model rats,delaying articular cartilage degeneration,and thus improve the symptoms of KOA.