尼古丁对白细胞介素-1β诱导的软骨细胞炎性反应及核因子-κB信号通路的影响  

Effect of nicotine on the inflammatory response of chondrocytes induced by interleukin-1β and nu- clear factor kappa B the signal pathway

作　　者：窦浚峰[1] 海国栋[1] 封青川[2] 张英娴[3] 白晓枫[4] Dou Xunfeng;Hai Guodong;Feng Qingchuan;Zhang Yingxian;Bai Xiaofeng(Department of Surgical Bone Tumour and Osteopathy, Zhengzhou Orthopaedics Hospital, Zhengzhou 450052, China;Department of Cell Biology and Medical Genetics, Basic Medical College, Zhengzhou University, Zhengzhou 450052, China;Department of Endocrinology and Metabolism, Zhengzhou Children＇ s Hospital, Zhengzhou 450053, China;Department of Abdominal Surgery, Cancer Hospital Chinese Academy of Medical Sciences, Belling 100021, China)

机构地区：[1]郑州市骨科医院骨肿瘤骨病外科,450052 [2]郑州大学基础医学院细胞生物学与医学遗传学系,450052 [3]郑州市儿童医院内分泌代谢科,450053 [4]中国医学科学院肿瘤医院腹部外科,北京100021

出　　处：《中华实验外科杂志》2018年第5期885-887,共3页Chinese Journal of Experimental Surgery

摘　　要：目的 观察尼古丁对白细胞介素-1β（IL-1β）诱导的软骨细胞炎性反应及核因子-κB（NF-κB）信号通路的影响.方法 通过Ⅱ型胶原酶消化分离大鼠原代软骨细胞,并利用噻唑蓝（MTF）法检测10-8、10-7、10-6、10-5 mol/L尼古丁对软骨细胞活力的影响.随后软骨细胞分为5组,分别为正常组（不含任何药物刺激）,模型组（10 μmol/L IL-1β）,尼古丁低、中、高剂量组（10-8、10-7、10-6 mol/L＋ 10μmol/L IL-1β）.MTT法检测各组细胞活力,酶联免疫吸附试验（ELISA）检测肿瘤坏死因子-α（TNF-α）,白细胞介素-γ（IFN-γ）及IL-6含量,Western blot检测各组细胞中诱导型一氧化氮合酶（iNOS）、p-NF-κB p65及p-IκBα表达量.结果 与正常组比较,10-8、10-7、10-6 mol/L尼古丁（0.52 ±0.05、0.64±0.06、0.78±0.07）显著提高软骨细胞活力（P=0.032、0.004、0.000）.与正常组比较,模型组中软骨细胞活力降低,TNF-α、IFN-γ及IL-6含量[（8.73±0.84）、（17.24±1.76）、（2.78 ±0.21） ng/L]提高（P =0.002、0.001、0.001）,iNOS、p-NF-κB p65及p-IκBα表达量上调.与正常组比较,尼古丁低、中、高剂量组中细胞活力提高,TNF-α、IFN-γ及IL-6含量降低[低剂量组（6.61±0.14）、（15.36±1.33）、（2.29±0.19） ng/L,P=0.004、0.000、0.000;中剂量组（5.52±0.50）、（13.23±1.27）、（1.82±0.17） ng/L,P=0.007、0.000、0.000;高剂量组（3.93±0.40）、（10.45±1.04）、（1.54±0.15） ng/L,P=0.005、0.000、0.000],p-IκBα表达量下调,尼古丁中、高剂量组中iNOS及p-NF-κB p65表达量显著下调（中剂量组：0.38±0.04、0.40±0.04,P=0.008、0.000;高剂量组：0.31±0.30、0.30±0.05,P=0.000、0.000）.结论 一定剂量尼古丁能通过抑制NF-κB信号通路来抑制IL-1β诱导的软骨细胞炎性反应.Objective To explore effect of nicotine on the inflammatory response of chondrocytes induced by interleukin-1 β （IL-1 β） and nuclear factor kappa B （NF-κB） signal pathway.Methods Rat primary chondrocytes were isolated by collagenase digestion.Effect of 10-8,10-7,10-6,10-5 mol/L nicotine on chondrocytes viability was detected by methyl thiazol tetrazolium （MTF）.Chondrocytes were divided into 5 groups,normal group （without any drug stimulation）,model group （10 μmol/L IL-1β）,nicotine low,middle and high dose group （l0-8,10-7,10-6 mol/L ＋ 10 μmol/L IL-1β）.The viability of chondrocytes was detected by MTF assay.The content of tumor necrosis factor alpha （TNF-α）,interleukin gamma （IFN-γ） and IL-6 was detected by enzyme linked immunosorbent assay （ELISA）.The expression of nitric oxide was detected （iNOS）,p-NF-κB p65 and p-IκBα was detected by Western blotting.Results Compared with normal group,10-8,10-7,10-6 mol/L nicotine [（0.52 ±0.05）,（0.64 ± 0.06）,（0.78 ± 0.07）] increased chondrocytes＇ viability （P =0.032,0.004,0.000）.Compared with normal group,cell viability was decreased （P =0.000）,the level of TNF-α,IFN-γ and IL-6wasincreased [（8.73 ±0.84）,（17.24 ±1.76）,（2.78 ±0.21） ng/L] （P=0.002,0.001,0.001）,the expression of iNOS,p-NF-κB p65 and p-IκBα was up-regulated in model group（P =0.003,0.000,0.000）.Compared with model group,cell viability was decreased,（P =0.009,0.000,0.000） the level of TNF-α,IFN-γ and IL-6 was reduced [（6.61 ±0.14）,（15.36±1.33）,（2.29±0.19） ng/L,P =0.004,0.000,0.000;（5.52 ±0.50）,（13.23 ±1.27）,（1.82 ±0.17） ng/L,P =0.007,0.000,0.000;（3.93 ±0.40）,（10.45 ±1.04）,（1.54 ±0.15） ng/L,P =0.005,0.000,0.000],the expression of p-IκBα （P =0.009,0.001,0.000） was down-regulated in nicotine low,middle and high dose group,the expression of iNOS and p-NF-κB p65 was down-regulated in middle and high dose group （0.38 ±0.04,0.40 ±0.04,P=0.008,0.000;0.31 ±0.3

关 键 词：尼古丁 白细胞介素-1Β 软骨细胞 炎症 核因子-ΚB信号通路 

分 类 号：R681.3[医药卫生—骨科学]