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1、血小板源性生长因子BB表达对促进大鼠肌腱愈合和防止肌腱粘连的作用南京大学医学院附属鼓楼医院 林 樾 梁红伟 李云剑 燕 辛 谭 谦【摘要】目的 研究血小板源性生长因子BB（PDGF-BB）基因转染大鼠肌腱细胞后促进肌腱愈合及防止肌腱粘连的作用。 方法 90只大鼠随机分为A、B、C三组，建立跟腱损伤模型。A组：实验组，肌腱断端注射PDGF-BB基因转染的肌腱细胞；B组：对照组，肌腱断端注射肌腱细胞；C组：空白对照组。6-0丝线行改良Kessler法缝合跟腱，管型石膏固定一周。术后第3d、1w、2w、4w、8w取样，分别行大体、组织学观察、力学以及免疫学检测，对比肌腱粘连度、组织中成纤维细胞数量、
2、胶原纤维含量、肌腱最大抗拉力及最大滑动距离、组织中PDGF-BB的浓度。结果 PDGF-BB基因转染肌腱细胞经过RT-PCR以及测序证实在体外稳定表达。实验组在肌腱粘连度、胶原纤维数量、成纤维细胞数量、肌腱最大滑动距离以及组织中PDGF-BB含量等方面均优于对照组（P0.05或P0.05)。结论 运用PDGF-BB基因转染肌腱细胞注射有促进肌腱内源性愈合、减轻肌腱粘连的作用。【关键词】PDGF-BB基因；肌腱愈合；肌腱粘连；基因转染【Abstract】Objective To study the effect of platelet-derived growth factor-BB（PDGF-
3、BB）gene expression in the process of promoting rat tendon healing and preventing adhesion of rat tendon.Methods 90 rats were randomly divided into three groups (gorup A、B and C), established tendon injury models. Group A: injected PDGF-BB gene transfected rat tendon cells in the end of injury tendon
4、; Group B: injected tendon cells; Group C: control. Then samples were taken from rats tendons in different times including 3 days,1 week,2 weeks,4 weeks and 8 weeks after operation. Morphology and histology observation、mechanical testing and immunoassay were given. The effect of PDGF-BB gene transfe
5、cted rat tendon cells in the process of promoting tendon healing and preventing adhesion of tendons were proved by comparing of the degree of tendon adhesion ;The number of tissue fibroblasts and collagen fiber content; tendon maximum tensile strength and the maximum sliding distance after the tendo
6、n healing ; levels of PDGF-BB in tendon tissue as well.Results The transfection of PDGF-BB gene into tendon cells were testified by RT-PCR and sequencing, and PDGF-BB could stably expressed in vitro. Compared with control, the PDGF-BB gene transfected rat tendon cells had more advantages in the degr
7、ee of tendon adhesion; The number of tissue fibroblasts and collagen fibers; the maximum sliding distance and the content of PDGF-BB in tendon tissue（P0.05 or P0.05)(表1)。图1 RT-PCR检测PDGF-BB mRNA 的表达：1. marker 2.转染组RT-PCR3.转染组-actin RT-PCR 4.未转染组RT-PCR 5.未转染组-actin RT-PCR表1术后8周各组肌腱粘连程度分级(n=10)组别肌腱粘连分级
8、级级级级级A组01231B组11233C组012253.组织学观察： 术后3天各组肌腱断端均可见充血水肿及炎细胞浸润；术后1周，充血、水肿较术后3天有所减少，同时胶原纤维增生，炎性细胞向胶原纤维浸润，并见吞噬细胞活跃；腱周组织可见毛细血管增生、成纤维细胞增殖活跃，其中以C组最明显；A组细胞数量相对较少（P0.05），而B、C组近断端的腱实质内可见较多成纤维细胞增殖，断端有新生胶原形成，并出现较多的新生毛细血管。术后2-4周，各组肌腱的炎性反应逐渐消退，成纤维细胞增殖活跃并向肌腱断端迁移，可见排列紊乱的胶原成分和毛细血管增生。术后8 周, 各组成纤维细胞数较前明显减少，组间数量趋于一致。肌腱断端胶原纤维的排列方向趋于一致，与肌腱纵轴平行，肌腱与周围组织的粘连较前减轻。愈合过程中B、C组各部位的成纤维细胞及胶原纤维含量均较A组多（P0.0
