多配体蛋白聚糖4在下颌髁突软骨退变中作用的初步研究  

A preliminary study on the role of syndecan4 in the degeneration of mandibular condyle cartilage

作　　者：陈晓华 牟婧 何睿 何峰 马原军 吴凡 段宇辰 苗辉 于世宾 CHEN Xiaohua;MOU Jing;HE Rui;HE Feng;MA Yuanjun;WU Fan;DUAN Yuchen;MIAO Hui;YU Shibin(State Key Laboratory of Military Stomatology,National Clinical Research Center for Oral Diseases,Shaanxi International Joint Research Center for Oral Diseases,Department of Oral Anatomy and Physiology and TMD,The Third Hospital Affiliated to Air Force Military Medical University,Xi'an,710032,China;Lintong Medical Rehabilitation Center,Xi'an)

机构地区：[1]军事口腔医学国家重点实验室,国家口腔疾病临床医学研究中心,陕西省口腔疾病国际联合研究中心,空军军医大学第三附属医院解剖生理学教研室,西安710032 [2]临潼康复疗养中心

出　　处：《实用口腔医学杂志》2023年第2期153-159,共7页Journal of Practical Stomatology

基　　金：国家自然科学基金面上项目(编号:81970953,82271000);陕西省自然科学基础研究计划重点项目(编号:2022JZ-52);陕西省重点研发计划项目(编号:2021SF-046)。

摘　　要：目的:初步探究多配体蛋白聚糖4(SDC4)在下颌髁突软骨(MCC)退变中的作用。方法:C57小鼠随机分为对照组(Con)和单侧前牙反组(UAC)(n=6)。3周后番红O-固绿、HE染色检测MCC退变程度;免疫组化检测MCC中SDC4、降解因子MMP13、TNFα以及细胞凋亡关键蛋白cleaved-CASP3的表达。体外培养SD大鼠MCC原代细胞,分别施加30%、0.5 Hz循环拉应力6 h(T6 h组)、12 h(T12 h组),Western blot、RT-qPCR等检测SDC4、ADAMTS5、TNFα以及cleaved-CASP3的表达变化。力学加载情况下向细胞培养上清中加入SDC4胞外段抗体(T30%+Anti-SDC4组)或大鼠IgG抗体(T30%+IgG组),检测上述细胞因子表达变化。结果:与Con组相比,UAC组MCC厚度、番红O阳性面积比降低(P<0.01、P<0.05);SDC4、MMP13、TNFα、cleaved-CASP3阳性细胞率升高(P<0.05、P<0.01、P<0.05、P<0.01)。与Con6 h、Con12 h组比较,T6 h、T12 h组MCC细胞SDC4、TNFα、cleaved-CASP3的蛋白表达升高(P<0.01)。与T30%+IgG组相比,加入Anti-SDC4抗体可逆转应力所导致的ADAMTS5、TNFα、cleaved-CASP3的表达上调(P<0.05、P<0.01、P<0.01)。结论:SDC4可能在MCC退变中发挥着重要作用。Objective:To explore the role of syndecan4(SDC4)in the degeneration of mandibular condylar cartilage(MCC).Methods:C57 mice were randomly divided into control(Con)and unilateral anterior crossbite(UAC)groups(n=6),and were sacrificed 3 weeks later.HE and Safranin O staining were performed to evaluate the degree of MCC degeneration.Immunohistochemical staining was used to detect the expression of SDC4,catabolic cytokines MMP13,TNFα,and chondrocytes apoptosis-key protein cleaved-CASP3 in cartilage.In vitro cultured primary MCC chondrocytes from SD rats were treated with 0.5 Hz and 30%cyclic tensile stress loading for 6 and 12 h(T6 h and T12 h groups).Western blot and RT-qPCR were performed to detect the expression of SDC4,ADAMTS5,TNFαand cleaved-CASP3.Then,on the basis of the cyclic tensile strain,anti-SDC4 extracellular antibody(T30%+anti-SDC4 group)or rat IgG(T30%+IgG group)was added in the cell culture medium.The expression of above-mentioned cytokines was detected.Results:Compared with the Con group,the cartilage thickness and Safranin O-positive area ratio in the UAC group were decreased(P<0.05 and P<0.01),the SDC4,MMP13,TNFαand cleaved-CASP3-positive cells were increased(P<0.05,P<0.01,P<0.05 and P<0.01).Compared with the Con6 h and Con12 h groups,the protein expression of SDC4 and TNFα,cleaved-CASP3 in the MCC chondrocytes was increased in the T6 h and T12 h groups(P<0.01).The Anti-SDC4 antibody reversed the up-regulated expression level of ADAMTS5,TNFαand cleaved-CASP3(P<0.05,P<0.01 and P<0.01).Conclusion:SDC4 may play an impor tant role in the degeneration of MCC.

关 键 词：髁突 软骨 骨关节炎 多配体蛋白聚糖4 拉应力 

分 类 号：R782.6[医药卫生—口腔医学]