基于NLRP3炎症小体活化探讨“三色散”挥发油减轻膝骨关节炎滑膜炎症的机制
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篇名: 基于NLRP3炎症小体活化探讨“三色散”挥发油减轻膝骨关节炎滑膜炎症的机制
TITLE: Discussion on the Mechanism of Synovitis of KOA Relieved by “Sanse Powder ”Volatile Oil Based on the Activation of NLRP 3 Inflammasome
摘要: 目的:基于NOD样受体家族3(NLRP3)炎症小体的活化探讨“三色散”挥发油减轻膝骨关节炎(KOA)滑膜炎症的机制。方法:采取水蒸气蒸馏法提取“三色散”挥发油,采用气质联用(GC-MS)技术分析其组成。提取雄性SD大鼠膝关节成纤维样滑膜细胞(FLS),采用CCK-8法检测10、25、50、100、250、500、1000μg/mL“三色散”挥发油对FLS存活率的影响;以脂多糖诱导12h建立KOA炎症细胞模型,检测10、250μg/mL“三色散”挥发油对炎症模型细胞中NLRP3、胱天蛋白酶1(caspase-1)、凋亡相关斑点样蛋白(ASC)的蛋白及其mRNA相对表达量的影响,检测炎症模型细胞上清液中白细胞介素1β(IL-1β)、IL-18水平。结果:提取得到黄色澄清且有独特芳香气味的“三色散”挥发油0.51~0.61g,提取率为0.33%~0.41%(n=9)。从该挥发油中共分离得到化学成分41种,鉴定了其中的30种,其峰面积之和占总峰面积的90.0736%;相对含量较高的成分依次为芳姜黄酮(17.5739%)、δ-杜松烯(15.4345%)、姜黄酮(11.5095%)等。当“三色散”挥发油质量浓度为10~250μg/mL时,其对FLS存活率均无显著影响(P>0.05)。与空白组比较,模型组细胞中NLRP3、caspase-1、ASC的蛋白及其mRNA相对表达量以及细胞上清液中IL-1β、IL-18水平均显著升高(P<0.05);与模型组比较“,三色散”挥发油10、250μg/mL组细胞及上清液中上述指标的相对表达量或水平均显著降低(P<0.05)。结论:“三色散”挥发油可能通过抑制FLS中NLRP3炎症小体的活化而减少下游炎症级联反应,从而发挥其改善KOA滑膜炎症的作用。
ABSTRACT: OBJECTIVE:To discuss the mechanism of synovitis of knee osteoarthritis (KOA)relieved by “Sanse powder ” volatile oil based on the activation of Nod-like receptor family 3(NLRP3)inflammasome. METHODS :“Sanse powder ”volatile oil was extracted by distillation- condensation process and the components of it were analyzed by GC-MS. Fibroblast-like synovial cells(FLS)were extracted from the knee joint of male SD rats. The effects of 10,25,50,100,250,500,1 000 μg/mL“Sanse powder”volatile oil on the viability of FLS were examined by CCK-8 assay. KOA inflammatory cell model was induced by lipopolysaccharide for 12 h. The effects of 10,250 μg/mL“Sanse powder ”volatile oil on the protein and mRNA expression of NLRP3,caspase-1 and apoptosis-associated speck-like protein (ASC)were detected. The levels of IL- 1β and IL-18 in supernatants of FLS were determined . RESULTS :Totally 0.51-0.61 g volatile oil with yellow clear and unique aromatic odor were extracted from“Sanse powder ”,and the extraction rate was 0.33%-0.41%(n=9). A total of 41 chemical components were isolated ,and 30 of them were identified and their peak area accounted for 90.073 6% of the total peak area. The components with high relative content were gingerol flavonoids (17.573 9%),δ-junionene(15.434 5%),gingerol(11.509 5%),etc. When the concentration of volatile oil was 10-250 μg/mL,there was no significant effect on survival rate of FLS (P>0.05). Compared with blank group ,the relative protein and mRNA expression of NLRP 3,caspase-1 and ASC and the levels of IL- 1β and IL-18 in supernatant were significantly increased in model group (P<0.05). Compared with model group ,relative expression or levels of above indexes were all decreased significantly in FLS and supernatant of “Sanse powder ”volatile oil 10 and 250 μg/mL groups(P<0.05). CONCLUSIONS:“Sanse powder ”volatile oil can inhibit NLRP 3 inflammasome activation in FLS and reduce the downstream inflammatory cascade response ,thus exerting its efficacy in ameliorating synovial inflammation of KOA.
期刊: 2021年第32卷第19期
作者: 廖太阳,张力,张皞晟,李晓辰,吴鹏,王培民
AUTHORS: LIAO Taiyang ,ZHANG Li,ZHANG Haosheng ,LI Xiaochen ,WU Peng,WANG Peimin
关键字: 膝骨关节炎;滑膜炎症;NOD样受体家族3炎症小体;三色散;挥发油;成纤维样滑膜细胞
KEYWORDS: Knee osteoarthritis ;Synovial inflammation ;Nod-like receptor family 3 inflammasome;Sanse powder ;Volatile
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