17β-雌二醇对白细胞介素1-β诱导的软骨细胞损伤的抑制作用  

Inhibitory effect of 17β-estradiol on chondrocyte injury induced by interleukin-1β

作　　者：王琮仁[1] 郭祥[1] 金旭红[1] 吴多庆[1] 孔长庚[1] Wang Congren;Guo Xiang;Jin Xuhong;Wu Duoqing;Kong Changgeng(Orthopaedic Medical Center,Haikou Hospital,Xiangya Medical College,Central South University,Haikou 572000,China)

机构地区：[1]中南大学湘雅医学院附属海口医院骨科医学中心,海口572000

出　　处：《中华实验外科杂志》2023年第7期1250-1252,共3页Chinese Journal of Experimental Surgery

摘　　要：目的探讨17β-雌二醇(E2)对白细胞介素1-β(IL-1β)诱导的软骨细胞损伤的抑制作用。方法酶消化法获得软骨细胞,并将细胞分为5组,正常组(含有10%胎牛血清的培养基);IL-1β组(10 ng/ml的IL-1β);1×10^(-9)mol/L组(10 ng/ml的IL-1β+1×10^(-9)mol/L E2);1×10^(-8)mol/L组(10ng/ml的IL-1β+1×10^(-8)mol/LE2);1×10^(-7)mol/L组(10ng/ml的IL-1β+1×10^(-7)mol/LE2)。噻唑蓝(MTT)法检测细胞活力,膜联蛋白V(AnnexinV)-碘化丙锭(PI)流式双染法检测细胞凋亡情况,酶联免疫吸附法(ELISA)法检测前列腺素E2(PGE2)、软骨寡聚基质蛋白(COMP)硫酸软骨素846(CS846)表达,蛋白质印迹法(Westernblot)检测细胞中环氧化酶-2(COX-2)、B淋巴细胞瘤-2基因(bcl-2)、bcl-2相关X蛋白(bax),基质金属蛋白酶-3(MMP-3),MMP-13表达及核因子κB(NF-κB)信号通路激活情况。组间比较采用单因素方差分析。结果1×10^(-9)、1×10^(-8)、1×10^(-7)mol/L组细胞活力、bcl-2表达量高于IL-1β组(0.53±0.05、0.61±0.06、0.69±0.06比0.78±0.07,t=6.254,P<0.01),(0.36±0.04、0.68±0.07、1.23±0.12比0.30±0.03,t=5.210,P<0.05),1×10^(-1)×10^(-8)、1×10^(-7)mol/L组细胞早期凋亡率、晚期凋亡率、PGE2含量、COMP含量、CS846含量、COX-2表达量、MMP-13表达量及p-NF-κBp65低于IL-1β组[(6.52±0.66)%、(13.26±1.43)%、(18.54±1.85)%比(23.59±2.54)%,t=6.324,P<0.01]、[(4.23±0.41)%、(6.87±0.69)%、(10.51±1.06)%比(15.84±1.58)%,t=6.746,P<0.01]、[(23.69±2.80)(36.23±3.62)、(48.21±4.82)ng/L比(59.82±5.98)ng/L,t=5.326,P<0.01]、[(0.20±0.02)、(0.35±0.03)、(0.48±0.05)μg/L比(0.62±0.06)μg/L,t=6.329,P<0.01]、[(0.35±0.03)(0.48±0.05)、(0.65±0.03)ng/L比(0.78±0.08)ng/L,t=6.534,P<0.01]、[(0.12±0.01)、(0.04±0.04)、(0.42±0.04)比(1.02±0.10),t=6.359,P<0.01]、[(0.25±0.02)(0.30±0.03)、(0.65±0.07)比(1.02±0.13),t=5.984,P<0.01]、[(0.32±0.03)、(0.42±0.04)、(0.97±0.09)比(1.29±0.13),t=5.364,P<0.01],1×10^(-8)、1×10^(-7)mol/L组bax及MMP-3表达量低于IL-1β组[(0.76±0.07)�Objective To explore the inhibitory effect of 17β-estradiol(E2)on chondrocyte injury induced by interleukin-1β(IL-1β).Methods Chondrocytes were obtained by enzyme digestion and randomly divided into 5 groups:normal group(medium containing 10%fetal bovine serum);IL-1βgroup(10 ng/ml IL-1β),1×10^(-9) mol/L E2 group(10 ng/ml IL-1β+1×10^(-9)mol/L E2),1×10^(-8)mol/L E2 group(10 ng/ml IL-1β+1×10^(-8) mol/L E2),1×10^(-7) mol/L E2 group(10 ng/ml IL-1β+1×10^(-7) mol/L E2).Cell viability was detect by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bro-mide(MTT)assay.Cell apoptosis was detected by Annexin V-propidium iodide(PI)flow cytometry.The expression of prostaglandin E2(PGE2),cartilage oligomeric matrix protein(COMP)and chondroitin sulfate(CS846)was detected by enzyme linked immunosorbent assay(ELISA).The expression of cycloox-ygenase-2(COX-2),B-cell lymphoma-2(bcl-2),bcl-2-associated X protein(bax),matrix metalloprotein-ase(MMP)-3,MMP-13 and the activation of nuclear factor kappa-B(NF-kB)p65 signal pathway were de-tected by Western blotting.Results was analyzed by single-factor analysis of variance.Results The cell viability and the expression of bcl-2[(0.53±0.05),(0.61±0.06),(0.69±0.06)vs.(0.78±0.07),t=6.254,P<0.05],[(0.36±0.04),(0.68±0.07),(1.23±0.12)vs.(0.30±0.03),t=5.210,P<0.05]in 1×10^(-9),1×10^(-8),1×10^(-7) mol/L E2 groups were significantly higher than those in IL-1βgroup.The early apoptosis rate,late apoptosis rate,the level of PCE2,COMP,CS846,COX-2,MMP-13 and p-NF-kB p65[(6.52±0.66)%,(13.26±1.43)%,(18.54±1.85)%vs.(23.59±2.54)%,t=6.324,P<0.01],[(4.23±0.41)%,(6.87±0.69)%,(4.23±0.41)%vs.(15.84±1.58)%,t=6.746,P<0.01][(23.69±2.80),(36.23±3.62),(48.21±4.82)ng/L vs.(59.82±5.98)ng/L,t=5.326,P<0.01],[(0.20±0.02),(0.35±0.03),(0.48±0.05)vs.(0.62±0.06)μg/L,t=6.329,P<0.01],[(0.35±0.03),(0.48±0.05),(0.65±0.03)ng/L vs.(0.78±0.08)ng/L,t=6.534,P<0.01],[(0.12±0.01),(0.04±0.04),(0.42±0.04)vs.(1.02±0.10),t=6.359,P<0.01],[(0.25±0.02),(0.30±0.03),(0.65±0.07)vs.(1.02±0.1

关 键 词：17Β-雌二醇 骨关节炎 软骨细胞 凋亡 细胞外基质 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]