抑制瘢痕细胞中串珠素表达对Ⅰ型胶原蛋白生成量的影响(英文)  

The Influence of the TypeⅠCollagen Generation by Interference Perlecan Express

作　　者：孙明岳[1] 褚言琛[1] 迟静薇[1] 孙菲[1] 邹云雯[1] 

机构地区：[1]青岛大学,山东青岛266003

出　　处：《现代生物医学进展》2013年第14期2650-2656,共7页Progress in Modern Biomedicine

基　　金：Natural Science Foundation of China(81071482)~~

摘　　要：目的:硬膜外瘢痕,又叫硬膜外纤维化,是指在硬膜外腔的手术涉及范围内形成的瘢痕组织或纤维化,是机体对创伤的修复反应。瘢痕的粘连和收缩会牵拉硬膜和神经根,限制其活动,被瘢痕包绕的神经根受到非正常的牵拉和挤压,神经纤维的轴浆运输、动脉血供、静脉回流受阻,神经根和背侧神经节对机械压迫很敏感,会产生一系列症状,如疼痛、麻木及下肢肌力降低等。近年来,对硬膜外瘢痕防治的研究大多是椎板切除术后如何通过物理或化学屏障来减少术后因瘢痕粘连导致的并发症。但对通过瘢痕形成过程中抑制其主要构成成分的生成来减轻椎板切除术后硬膜外瘢痕形成的相关研究还较少。通过减少椎板切除术后硬膜外瘢痕主要成分Ⅰ型胶原蛋白的生成来实现抑制椎板切除术后硬膜外瘢痕的形成。方法:选用30只250克两月鼠龄的SD雄性大鼠随机按1、2、3、4、5、6周分为6组,行后路4、5腰椎全椎板切除术。术后1、2、3、4、5、6周时每周取一组大鼠全锥板切除术后硬膜后方瘢痕组织,分别行病理切片HE染色,组织块贴壁法细胞培养。筛选第三周瘢痕组织培养的成纤维细胞进行慢病毒干扰串珠素表达并设对照组,通过Western-blot、RT-PCR分析Ⅰ型胶原蛋白生成量与对照组的差别并进行统计学分析。结果:慢病毒干扰小组Ⅰ型胶原蛋白生成量较对照组及纯病毒组明显减少(RT-PCR F=509.331,q A,B=-43.371,P<0.01,q A,C=-46.133,P<0.01,Western-Blot F=337.578;q A,B=-112.433,P<0.01,q A,C=-89.227,P<0.01)。结论:干扰串珠素表达能有效减少术后硬膜外瘢痕成纤维细胞生成Ⅰ型胶原蛋白,对抑制椎板切除术后硬膜外瘢痕形成应有一定作用。通过慢病毒介导的shRNA干扰成纤维细胞中的串珠素后,其生成的Ⅰ型胶原蛋白量与对照组相比较差异有统计学意义(P<0.05),这说明通过抑制瘢痕成纤维细胞的串珠素表达能�Objective： Epidural scar,also known as epidural fibrosis,means scar tissue or fibrosis formed in the surgery within the scope of the epidural,and it is body＇s trauma repair response.Scar adhesion and contraction will stretch the dura mater and nerve root,limit their activities,scar encysted nerve root may be abnormal stretched and squeezed,nerve fiber＇s axoplasmic transportation,arterial blood supply and venous return suffocated,nerve root and dorsal ganglion is very sensitive to mechanical pressure.Therefore,it will produce a series of symptoms,such as pain,numbness,lower extremity muscle strength is reduced.Recently,most of the research on the prevention of epidural scar is how to reduce the occurrence of postoperative scar adhesions（which leads to the occurrence of complications） through physical or chemical barrier after the laminectomy.But,the research on inhibit scar＇s main constituent in the process of scar information to reduce epidural scar formation after laminectomy is few.Inhibition epidural scar formation by reducing typeⅠ collagen production.Methods： 30 SD rats were randomly and evenly divided into six groups and full laminectomy was performed on the 4th,5th lumbar vertebra in each experiment.The epidural scar were sampled at the 1st,2nd,3rd,4th,5th and 6th weeks postoperatively,one group each week.Then hematoxylin-eosin staining（HE） was done to observe the formation of epidural scar and the activity of desmocyte and the quantity of collagen synthesis.Cell culture was done to observe the desmocyte morphology and activity.Western Blot and RT-PCR were made to quantify the expression of the perlecan and type I collagen after interference by lentivirus mediated shRNA interference.Results： According to hematoxylin-eosin staining（HE） and cell culture,scar desmocytes at the second and third weeks are the most suitable for the experiment.Western Blot and RT-PCR showed that type I collagen expression reduced compared to control group after the perlecan was interference by lentivirus

关 键 词：椎板切除术 硬膜外瘢痕 基底膜聚糖 胶原蛋白 纤维细胞 

分 类 号：R62[医药卫生—整形外科] R68[医药卫生—外科学]