CO2激光犬声带全切除后不同时间细胞外基质表达  

Expression of Extracellular Matrix at Different Periods after CO 2 Laser Complete Removal of Vocal Cords in Canine

作　　者：汪斌如[1] 张莹莹[1] 杨丽萍[1] 刘莉[1] 陈始明[2] 梁耕田[1] Wang Binru;Zhang Yingying;Yang Liping;Liu Li;Chen Shiming;Liang Gengtian(Department of Otolaryngology Head and Neck Surgery,Tongren Hospital of Wuhan University(Wuhan Third Hospital),Wuhan,430060,China)

机构地区：[1]武汉大学附属同仁医院(武汉市第三医院)耳鼻咽喉头颈外科,武汉430060 [2]武汉大学人民医院耳鼻咽喉头颈外科

出　　处：《听力学及言语疾病杂志》2020年第6期669-673,共5页Journal of Audiology and Speech Pathology

基　　金：湖北省自然科学基金项目(2018CFB455);湖北省卫生健康委员会科研项目(WJ2019M007)。

摘　　要：目的观察犬声带损伤后不同自愈时间段细胞外基质(extracellular matrix,ECM)相关影响因子的动态变化规律,探讨声带损伤后的可能修复机制。方法 5只实验犬分为实验组(4只,8侧声带)和对照组(1只,2侧声带)。实验组在支撑喉镜下暴露双侧声带,CO_2激光6焦耳/秒行双侧声带全切除深达肌层,对照组在支撑喉镜下观察声带形态后不作任何处理。实验犬于双侧声带CO_2激光损伤前及损伤后6 h、3 w、8 w和12 w分别进行喉镜检查声带创面愈合情况,然后,每个时间点取1只犬的声带标本行HE染色、免疫荧光染色检查,同时检测ECM中赖氨酰氧化酶(LOX)、基质金属蛋白酶-1(MMP-1)和缺氧诱导因子-1α(HIF-1α)的累积光密度值(IOD值)。结果 (1)喉镜检查示,术后3 w实验组犬声带创面仍有充血水肿,术后8 w声带创面光滑凹陷,术后12 w声带局部挛缩。(2)HE染色显示,术后6 h实验组声带创面大量红细胞和炎性细胞浸润;术后3 w创面成纤维细胞增生,新生血管形成;术后8 w创面纤维组织增生紊乱;术后12 w声带创面胶原纤维大量增生,排列紊乱,间隙增大。(3)免疫荧光染色显示,术后6 h和3 w,实验组声带创面炎性细胞、血管内皮细胞和腺体内LOX、MMP-1和HIF-1α表达较强烈;术后8 w和12 w,创面阳性细胞逐渐减少,随着创面愈合其表达趋于稳定。(4)各时间点LOX和MMP-1的IOD值比较差异均有统计学意义(P<0.05);除术后12 w与术前比较外,其余各时间点间HIF-1α的IOD值差异均有统计学意义(P<0.05)。结论 CO_2激光犬声带全切除后12 w内,ECM中LOX、MMP-1和HIF-1α差异化表达可能与声带的创伤修复及瘢痕形成有关。Objective To study dynamic expressions of extracellular matrix(ECM)related factors in different periods after vocal fold injuries in canine.Methods Five experimental dogs were divided into experimental group(4 dogs,8 vocal cords)and control group(1 dog,2 vocal cords).The experimental group was exposed to bilateral vocal cords under a supporting laryngoscope,and the CO 2 laser(6 J/s)were completely excised bilateral vocal cords to the deep muscular layer.The control group was treated with a laryngoscope to observe the vocal morphology without any treatment.Various experimental dogs were respectively performed laryngoscopy,HE staining,and immunofluorescence before and after 6 hours(6 h),3 weeks(3 w),8 w,and 12 w since CO 2 laser cauterization on dogs’bilateral vocal folds.Meanwhile,cumulative optical density value(IOD value)of lysyl oxidase(LOX),matrix metalloproteinase-1(MMP-1)and hypoxia-inducible factor-1α(HIF-1α)in ECM were tested.Results①Laryngoscopy indicated there were still congestion and edema on vocal fold wound surfaces after 3 w since operation.Wound surfaces became smooth and depressed after 8 w.There were local contractures after 12 w.②HE dyeing showed there was infiltration of many erythrocytes and inflammatory cells on wound surfaces after 6 h since operation.There were proliferation of fibroblasts and neovascularization on wound surfaces after 3 w.There were hyperplasia and derangement of fibrous tissues on wound surfaces after 8 w.Many collagenous fibers grew and were disorganized with larger clearances between tissues on wound surfaces at 12 w.③Immunofluorescence indicated LOX’s,MMP-1’s and HIF-1α’s expressions were intense in inflammatory cells,vascular endothelial cells,and glands on wound surfaces.LOX’s,MMP-1’s and HIF-1α’s expressions were strong in early and middle postoperative periods,and their expressions were stable with wound healing.④IOD values showed:LOX’s and MMP-1’s expressions differed at different periods(P<0.05).Their expressions before and after the

关 键 词：声带 声带损伤 细胞外基质 

分 类 号：R767.4[医药卫生—耳鼻咽喉科]