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12 h建立KOA炎症细胞模型,检测10、250μg/m L"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0.51~0.61 g,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/m L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。
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 10,25,50,100,250,500,1000μg/m L"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 10,250μg/m L"Sanse powder"volatile oil on the protein and m RNA expression of NLRP3,caspase-1 and apoptosis-associated speck-like protein(ASC)were detected.The levels of IL-1βand 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 90.0736%of the total peak area.The components with high relative content were gingerol flavonoids(17.5739%),δ-junionene(15.4345%),gingerol(11.5095%),etc.When the concentration of volatile oil was 10-250μg/m L,there was no significant effect on survival rate of FLS(P>0.05).Compared with blank group,the relative protein and m RNA expression of NLRP3,caspase-1 and ASC and the levels of IL-1βand 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 250μg/m L groups(P<0.05).CONCLUSIONS:"Sanse powder"volatile oil can inhibit NLRP3 inflammasome activation in FLS and reduce the downstream inflammatory cascade response,thus exerting its efficacy in ameliorating synovial inflammation of KOA.
作者
廖太阳
张力
张皞晟
李晓辰
吴鹏
王培民
LIAO Taiyang;ZHANG Li;ZHANG Haosheng;LI Xiaochen;WU Peng;WANG Peimin(Dept.of Orthopedics,the Affilliated Hospital of Nanjing University of Chinese Medicine/Jiangsu Province Hospital of Chinese Medicine,Nanjing 210029,China)
出处
《中国药房》
CAS
北大核心
2021年第19期2335-2341,共7页
China Pharmacy
基金
国家自然科学基金资助项目(No.81774334)
江苏省中医药领军人才培养项目(No.SLJ0207)。
关键词
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
Knee osteoarthritis
Synovial inflammation
Nod-like receptor family 3 inflammasome
Sanse powder
Volatile oil
Fibroblast-like synovial cells
