西红花酸对白细胞介素1β诱导人膝关节原代软骨细胞炎症因子表达的影响  

Effect of crocetin on expression of inflammatory factors in human chondrocytes of knee joints induced by interleukin-1β

作　　者：齐秀春[1] 陈昕[1] 曹玉净[1] 李扬[1] 宋鹏[1] 王前进[1] 艾进伟[1] 李浩亮[1] QI Xiu-chun;CHEN Xin;CAO Yu-jing;LI Yang;SONG Peng;WANG Qian-jin;AI Jin-wei;LI Hao-liang(Department of Trauma,Henan Province Hospital of TCM,Zhengzhou 450002,China)

机构地区：[1]河南省中医院创伤骨科,河南郑州450002

出　　处：《中国药理学与毒理学杂志》2020年第6期436-441,共6页Chinese Journal of Pharmacology and Toxicology

摘　　要：目的研究西红花酸对白细胞介素1β(IL-1β)诱导的人膝关节原代软骨细胞分解代谢及炎症应答的影响。方法西红花酸0.5,1,5,10和20μmol·L^-1处理人膝关节原代软骨细胞12 h,MTT检测细胞存活率。西红花酸0.5,1,5,10和20μmol·L^-1处理人膝关节原代软骨细胞12 h后,再加IL-1β10μg·L^-1处理24 h;实时荧光定量PCR(qRT-PCR)检测基质金属蛋白酶3(MMP-3)、MMP-13及Ⅱ型胶原(CollⅡ)mRNA水平;ELISA检测MMP-3、MMP-13、前列腺素E2(PGE2)、IL-6和肿瘤坏死因子α(TNF-α)水平;试剂盒检测一氧化氮(NO)含量;Western印迹法检测磷脂酰肌醇3激酶(PI3K)P85、磷酸化PI3K P85(p-PI3K P85)、蛋白激酶B(Akt)、p-Akt、P65 NF-κB和p-P65 NF-κB蛋白表达水平。结果MTT结果显示,西红花酸对人膝关节原代软骨细胞存活率无影响。qRT-PCR和ELISA检测结果显示,与IL-1β处理组相比,西红花酸处理可抑制IL-1β诱导的MMP-3和MMP-13 mRNA和蛋白表达(P<0.05),上调CollⅡmRNA表达水平(P<0.05)。西红花酸还可抑制IL-1β诱导的NO和PGE2释放(P<0.05),降低IL-1β处理组炎症因子IL-6和TNF-α的产生(P<0.05)。此外,IL-1β可显著上调人膝关节原代软骨细胞中p-Akt,p-P65 NF-κB,PI3K P85和p-PI3K P85蛋白表达,而西红花酸处理后可显著抑制IL-1β诱导的p-Akt,p-P65 NF-κB和p-PI3K P85蛋白表达(P<0.05)。结论西红花酸可抑制IL-1β诱导的人膝关节原代软骨细胞过度分解代谢及炎症应答,PI3K/Akt/NF-κB信号通路参与该过程,提示西红花酸可能具有潜在的抗骨关节炎的功效。OBJECTIVE To explore the role of crocetin in aberrant catabolism dysfunction and inflammation in interleukin-1β(IL-1β)-exposed chondrocytes.METHODS Human chondrocytes of knee joints were pretreated with crocetin 0.5,1,5,10 and 20μmol·L^-1 for 12 h.Cell viability was detected by MTT assay.Human chondrocytes were pretreated with crocetin 0.5,1,5,10 and 20μmol·L^-1 for 12 h and then treated with IL-1β(10μg·L^-1)for 24 h.The mRNA levels of matrix metalloproteinase-3(MMP-3),MMP-13 and collagenⅡwere determined by qRT-PCR.ELISA analysis was conducted to detect the contents of MMP-3,MMP-13,prostaglandin E2(PGE-2),IL-6 and tumor necrosis factor-α(TNF-α).Commercial kits were used to measure the levels of nitric oxide(NO)production.The protein expressions of phosphatidylinositol 3-kinase(PI3K)P85,phosphorylated PI3K P85(p-PI3K P85),Akt,p-Akt,P65 NF-κB and p-P65 NF-κB were evaluated by Western blotting.RESULTS No cytotoxicity was confirmed in crocetin-treated human chondrocytes of knee joints as detected by MTT.Administration with crocetin suppressed IL-1β-induced transcripts and productions of MMP-3 and MMP-13 relative to IL-1β-treated groups as detected by qRT-PCR and ELISA(P<0.05).Crocetin also upregulated the mRNA levels of collagenⅡ(P<0.05).Simultaneously,crocetin treatment restrained the release of NO and PGE2 in human chondrocytes of knee joints induced by IL-1β(P<0.05).Furthermore,IL-1β-evoked productions of inflammatory cytokine IL-6 and TNF-αin human chondrocytes of knee joints were abrogated by crocetin treatment(P<0.05).Additionally,exposure to IL-1βincreased the protein expressions of PI3K P85,p-PI3K P85,p-Akt and p-P65 NF-κB(P<0.05).The upregulated protein expressions of p-PI3K P85,p-Akt and p-P65 NF-κB were overturned when cells were administreed with crocetin(P<0.05).CONCLUSION Crocetin may inhibit IL-1β-evoked aberrant catabolism and inflammatory response in human chondrocytes of knee joints.PI3K/Akt/NF-κB signaling pathway is involved in this process.These results indicate the pot

关 键 词：西红花酸 骨关节炎 人膝关节原代软骨细胞 炎症因子 

分 类 号：R285[医药卫生—中药学] R967[医药卫生—中医学]