Siglec-9对TNF-α诱导的关节软骨细胞TLR4/NF-κB通路及凋亡的影响  

Effects of Siglec-9 on TLR4/NF-κB pathway and apoptosis of articular chondrocytes induced by TNF-α

作　　者：王晓元[1] 沈海丽[1] 王鑫[1] 张莉[1] 朱蓉[1] 熊咏民[2] WANG Xiaoyuan;SHEN Haili;WANG Xin(Department of Rheumatology and Immunology,The Second Hospital of Lanzhou University,Gansu,Lanzhou 730030,China)

机构地区：[1]兰州大学第二医院风湿免疫科,兰州市730030 [2]西安交通大学卫生科学中心公共卫生学院

出　　处：《河北医药》2022年第7期1014-1017,1021,共5页Hebei Medical Journal

摘　　要：目的探讨唾液酸结合性免疫球蛋白样凝集素9(Siglec-9)对肿瘤坏死因子-α(TNF-α)诱导的关节软骨细胞Toll样受体4(TLR4)/核因子κB(NF-κB)通路及凋亡的影响。方法体外培养人关节软骨细胞,分为对照组、TNF-α组(20 ng/ml TNF-α)、低浓度Siglec-9组(20 ng/ml TNF-α+2 ng/ml Siglec-9)、中浓度Siglec-9组(20 ng/ml TNF-α+10 ng/ml Siglec-9)和高浓度Siglec-9组(20 ng/ml TNF-α+50 ng/ml Siglec-9),MTT法检测软骨细胞活性,流式细胞仪检测软骨细胞凋亡情况,酶联免疫(ELISA)法检测软骨细胞上清液中白介素(IL)-1β、IL-6和IL-8水平,蛋白印迹(WB)法检测软骨细胞中TLR4、NF-κB p65、p-NF-κB p65、核因子κB抑制蛋白α(IκB-α)、p-IκB-α蛋白表达情况。结果与对照组相比,TNF-α组软骨细胞存活率显著降低(P<0.05),凋亡率、细胞上清液中IL-1β、IL-6、IL-8水平及细胞中TLR4、p-NF-κB p65/NF-κB p65、p-IκB-α/IκB-α蛋白水平显著升高(P<0.05);与TNF-α组相比,低、中、高浓度Siglec-9组软骨细胞存活率显著升高(P<0.05),凋亡率、细胞上清液中IL-1β、IL-6、IL-8水平及细胞中TLR4、p-NF-κB p65/NF-κB p65、p-IκB-α/IκB-α蛋白水平显著降低(P<0.05);随着Siglec-9处理浓度的升高,软骨细胞存活率显著升高,凋亡率、细胞上清液中IL-1β、IL-6、IL-8水平及细胞中TLR4、p-NF-κB p65/NF-κB p65、p-IκB-α/IκB-α蛋白水平显著降低(P<0.05)。结论Siglec-9对TNF-α诱导的关节软骨细胞TLR4/NF-κB通路激活及凋亡具有抑制作用。Objective To investigate the effects of sialic acid-binding Ig-like lectin 9(Siglec-9)on toll-like receptor 4(TLR4)/nuclear factor kappa B(NF-κB)pathway and apoptosis of articular chondrocytes induced by TNF-α.Methods Human articular chondrocytes were cultured in vitro and divided into control group,TNF-αgroup(20ng/ml),low concentration Siglec-9 group(20ng/ml TNF-α+2ng/ml Siglec-9),medium concentration Siglec-9 group(20ng/ml TNF-α+10ng/ml Siglec-9)and high concentration Siglec-9 group(20ng/ml TNF-α+50ng/ml Siglec-9).The activity of chondrocytes was detected by MTT assay,the apoptosis of chondrocytes was detected by flow cytometry,the levels of interleukin(IL)-1β,IL-6 and IL-8 in the supernatant were detected by enzyme-linked immunosorbent assay(ELISA),and Western Blot(WB)was used to detect the expression levels of TLR4,NF-κB p65,p-NF-κB p65,nuclear factor kappa B inhibitor protein alpha(IκB-α)and p-IκB-αin chondrocytes.Results Compared with those in control group,the survival rate of chondrocytes in TNF-αgroup was significantly decreased,however,the apoptosis rate,the levels of IL-1β,IL-6 and IL-8 in supernatant,and protein levels of TLR4,p-NF-κB p65/NF-κB p65 and p-IκB-α/IκB-αwere significantly increased(P<0.05).Compared with that in TNF-αgroup,the survival rate of chondrocytes in low,medium and high concentration Siglec-9 groups was significantly increased,however,the apoptosis rate,the levels of IL-1β,IL-6 and IL-8 in supernatant,protein levels of TLR4,p-NF-κB p65/NF-κB p65 and p-IκB-α/IκB-αwere significantly decreased(P<0.05).With the increase of Siglec-9 concentration,the survival rate of chondrocytes was increased significantly,but,the apoptosis rate,the levels of IL-1β,IL-6 and IL-8,and protein levels of TLR4,p-NF-κB p65/NF-κB p65 and p-IκB-α/IκB-αwere significantly decreased(P<0.05).Conclusion Siglec-9 can inhibit the activation of TNF-α-induced TLR4/NF-κB pathway and apoptosis of articular chondrocytes.

关 键 词：唾液酸结合性免疫球蛋白样凝集素9 肿瘤坏死因子-α 关节软骨细胞 TOLL样受体4 核因子ΚB 凋亡 

分 类 号：R684.3[医药卫生—骨科学]