骨桥蛋白-整合素ɑνβ3配体受体反应对人膝骨关节炎软骨细胞透明质酸表达水平的影响  

Osteopontin affects the expression of hyaluronic acid in human osteoarthritis chondrocytes by interacting with integrin avβ3 ligand receptor

作　　者：袁宇豪 罗伟[1] 刘擎 张方杰[3] Yuan Yuhao;Luo Wei;Liu Qing;Zhang Fangie(Department of Orthopedics,Niangya Hospital of Central South Unitersity,Changsha 410008,China;Department of Spine Surgery,the Second Xiangya Hospital of Central South University,Changsha 410011,China;Department of Emergency,Xiangya Hospital of Central South Unizersity,Changsha 410008,China)

机构地区：[1]中南大学湘雅医院骨科,长沙410008 [2]中南大学湘雅二医院脊柱外科,长沙410011 [3]中南大学湘雅医院急诊科,长沙410008

出　　处：《中国医师杂志》2020年第12期1782-1786,1791,共6页Journal of Chinese Physician

基　　金：国家自然科学基金青年基金(81501923);睿E急诊医学研究专项基金(R2019007)。

摘　　要：目的通过干预体外培养人膝骨关节炎软骨细胞中的骨桥蛋白(OPN)-整合素aB3配体受体反应,观察其对较骨细胞透明质酸(HA)表达水平的影响。方法从人膝骨关节炎软骨组织标本中获取并体外培养软骨细胞,实验分为三组:对照组加入1μg/ml重组人骨桥蛋白(rhOPN)干预;整合素ayβ3阻断组先采用整合素avB3阻断剂CRGDSP peptide(100μ/ml)干预软骨细胞1h以阻断OPN-整合素avβ3的结合后再加人1μ/ml thOPN进行干预23 h,整合素avβ3同型组先采用整合素av33同型剂GRGESP peptide 100μ/ml干预软骨细胞1h后再加入1μg/ml rhOPN进行干预23 h.采用实时荧光定量PCR(qPCR)法检测各组透明质酸合成酶HASI,HAS2和HAS3 mRNA表达,采用ELISA法检测HA水平。结果显微镜下观察膝骨关节炎软骨细胞与正常软骨细胞培养肩的形态,正常软骨细胞呈多角形,而膝骨关节炎软骨细胞伪足增多,形态多不规则。另外,与对照组(1.01±0.10)相比,整合素avB3阻断组HASI,HAS2,HAS3 mRNA表达水平[(0.70±0.01)、(0.68±0.05)、(0.55±0.03)]显著降低,差异有统计学意义(P<0.05);而整合素avβ3同型组HASI HAS2,HA53 mlRNA表达水平[(1.09±0.14)、(0.95±0.06).(1.04±0.08)]与对照组比较差异无统计学意义(P>0.05)。与对照组[(884.0±7.6)pg/ml]比较,整合素avβ3阻断组HA水平[(578.4±13.8)μg/ml]显著降低,差异有统计学意义(P<0.05);而整合素apβ3同型组HA水平[(882.1±10.1)μg/ml]与对照组比较差异无统计学意义(P>0.05)。结论OPN可以通过与整合素avB3配体受体反应刺激软骨细胞HAS分泌进而诱导HA表达。Objective To investigate whether osteopontin(OPN)-integrin avβ3 interaction could affect the expression of hyaluronic acid(HA)in human knee osteoarthritie chondrocytes in vitro by intervening its ligand-receptor interaction.Methods Chondroevtes were obtained from human osteoarthritis cartilage tissue and cultured in ritro.The chondrocytes were cultred in 6-well culture plates and divided into 3 groups:the control group were treated with 1μg/ml recombinant human osteopontin(rhOPN)for 23 hours;integrin avB3 blocking group were incubated with GRGDSP peptide(100μg/ml)for 1 hour before the rhOPN treated for 24 hours;integrin auB3 isotype control group were incubated with GRGESP peptide(100μg/ml)for 1 hour before the thOPN treated for 23 hours.Real-time-quantitative polymerase chain reaction(gRT-PCR)were used to determine the mRNA expression of the hyaluronic acid synthasel(HASI),HAS2 and HAS3.Enzyme-linked Immunosorbent assay(ELISA)were used to test the expression of HA.Results Under the microscope,the morphology of chondroeytes from knee osteoarthritis and normal chondroevtes were observed.The normal chondrocvtes were polygonal,while the pseudopodia of chondroeytes in knee osteoarthritis increased and the morphology was irregular.In addition,compared with the control group(1.01±0.10),the expression levels of HASI.HAS2 and HAS3 mRNA in the integrin avB3 blocking group[(0.70±0.01)、(0.68±0.05),(0.55±0.03)]were significantly lower than those in the control group(1.01±0.10),while there was no significant difference in the expression of HASI,HAS2 and HAS3 mRNA in the integrin avB3 isotype control group and control group[(1.09±0.14),(0.95±0.06),(1.04±0.08),P>0.05].Compared with the control group[(884.0±7.6)pg/ml],the expression of HA in integrin ay3 blocking group[(578.4±13.8)pg/ml]was significantly lower than that in the control group(P<0.05),but there was no significant difference in the HA expression level between the integrin ayB3 isotype control group[(882.1±10.1)μg/ml]and the control group(P>0.05).Co

关 键 词：骨桥蛋白质 骨关节炎 膝 软骨细胞 透明质酸 整合素aVβ3 

分 类 号：R684.3[医药卫生—骨科学]