文章摘要
张力,张立,邢润麟,黄正泉,李晓辰,徐波,肖延成,茆军,王培民.基于缺氧组织中NLRP3炎症小体的活化研究膝痹宁减轻KOA滑膜炎症的效应机制[J].南京中医药大学学报,2020,36(1):68-72.
基于缺氧组织中NLRP3炎症小体的活化研究膝痹宁减轻KOA滑膜炎症的效应机制
Mitigation Effect of Xibining on Synovitis of Knee Osteoarthritis Based on the Activation of NLRP3 Inflammasomes in Hypoxic Tissues
  
DOI:
中文关键词: 关键词:膝骨关节炎  滑膜炎症  膝痹宁  缺氧转录因子-1α  NLRP3炎症小体
英文关键词: knee osteoarthritis  synovitis  Xibining  HIF-1α  NLRP3 inflammasomes
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作者单位
张力,张立,邢润麟,黄正泉,李晓辰,徐波,肖延成,茆军,王培民 南京中医药大学附属医院/江苏省中医院江苏 南京 210029 
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中文摘要:
      目的 基于缺氧组织中NLRP3炎症小体的活化研究膝痹宁减轻膝骨关节炎滑膜炎症的效应机制。方法 大鼠分为空白组、KOA组、膝痹宁组。膝痹宁组药液灌胃,空白组、KOA组消毒生理盐水灌胃作为对照;第56天处死大鼠提取滑膜组织;HE染色评估滑膜炎症;缺氧探针免疫荧光染色观察大鼠膝关节滑膜组织缺氧程度;qPCR和Western blot检测缺氧转录因子1α(HIF-1α)、Caspase-1 p10、NLRP3及GSDMD的mRNA和蛋白表达;ELISA法测定NLRP3炎症小体活化下游IL-1β、IL-18的水平。结果 HE染色发现,KOA组滑膜组织较空白组表现出更多的炎性细胞浸润,衬里层细胞排列紊乱,膝痹宁组较KOA组炎性细胞浸润减少。大鼠滑膜组织的探针免疫荧光染色显示,KOA组与较空白组缺氧程度加重,膝痹宁组则较KOA组减轻。同时,KOA组滑膜组织中HIF-1α的mRNA和蛋白表达水平均较高于空白组(P<0.01),膝痹宁组则较KOA组降低(P<0.01)。此外,KOA组Caspase-1 p10、NLRP3、GSDMD的mRNA及蛋白表达水平均高于空白组(P<0.05),而膝痹宁组较KOA组有所降低(P<0.05)。KOA组滑膜组织中IL-1β、IL-18的水平较空白组升高(P<0.01),膝痹宁组较KOA组降低(P<0.01)。结论 膝痹宁能有效改善KOA滑膜组织缺氧状况,降低HIF-1α的表达,减少NLRP3炎症小体的活化,减轻滑膜炎症。
英文摘要:
      OBJECTIVE To explore the mitigation effect of Xibining on synovitis of knee osteoarthritis baesd on the activation of NLRP3 inflammasomes in hypoxic tissues. METHODS The rats were divided into three groups: normal group, KOA group and Xibining group. Xibining group was gavaged with drug liquid while the normal group and KOA group were treated with the same amount of sterilized saline as control. On the 56th day, the rats were killed to obtain synovial tissue. HE staining was performed to evaluate synovial inflammation and Hypoxyprobe hypoxia probe immunofluorescence staining was used to observe the degree of hypoxia in the synovial tissue of rats. qPCR and Western blot were performed to detect both gene and protein expression of HIF-1α, NLRP3, Caspase-1 p10 and GSDMD, respectively. The contents of IL-1β and IL-18, which were downstream from the activation of NLRP3 inflammasomes, were determined by ELISA. RESULTS Immunofluorescence staining with hypoxyprobe probe showed that the degree of hypoxia in synovial tissue of KOA rats was more severe than that of normal rats, and Xibining group was less than KOA. In HE staining, the synovial tissue of the KOA group showed more inflammatory cell infiltration and the lining layer cells were more disarranged than that of the normal group. The infiltration of inflammatory cells in the Xibining group was less than that in the KOA group. Meanwhile, both mRNA and protein level of HIF-1α in KOA group were higher than those in normal group (P<0.01), and Xibining group were lower than those in KOA group (P<0.01). In addition, both mRNA and protein expression of Caspase-1 p10, NLRP3 and GSDMD in KOA group were higher than those in normal group (P<0.05), while those in Xibining group were lower than those in KOA group (P<0.05). The contents of IL-1β and IL-18 in synovial tissue of KOA group was higher than that of normal group (P<0.01), and that of Xibining group was lower than that of KOA group (P<0.01). CONCLUSION Xibining can effectively improve the condition of hypoxia in synovial tissue of KOA, down-regulate the expression of HIF-1α, decrease the activation of NLRP3 inflammasomes, and alleviate synovitis.
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