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兔内侧副韧带和前交叉韧带正常及早期愈合过程中I型前胶原基因表达:一项原位杂交研究

Type I procollagen gene expression in normal and early healing of the medial collateral and anterior cruciate ligaments in rabbits: an in situ hybridization study.

作者信息

Wiig M E, Amiel D, Ivarsson M, Nagineni C N, Wallace C D, Arfors K E

机构信息

Division of Orthopaedics and Rehabilitation, University of California San Diego, La Jolla 92093-0630.

出版信息

J Orthop Res. 1991 May;9(3):374-82. doi: 10.1002/jor.1100090309.

Abstract

The purpose of this study was to compare the levels of procollagen type I messenger RNA (mRNA) in normal and healing medial collateral ligament (MCL) and anterior cruciate ligament (ACL) in a rabbit model. Our method of injury involved a surgical model with identical partial lacerations in the midsubstance of the MCL and ACL. Paraffin sections of normal ligaments, and ligaments 3, 7, 14, and 28 days postlaceration were studied by in situ hybridization to compare and follow the level of type I procollagen mRNA in the two ligaments. A complementary DNA (cDNA) probe corresponding to alpha 1(I) procollagen mRNA was labeled with [32P]d-CTP. After hybridization, autoradiography, and staining of the sections, the level of procollagen mRNA was assessed by microscopic examination. A higher level of procollagen mRNA was consistently detected in normal MCL than in normal ACL, suggesting higher collagen synthetic activity in the MCL. At the injury sites of the MCL and ACL, the levels of type I procollagen mRNA increased at all post-laceration periods, reaching its highest level at 14 days postsurgery. The MCL healing site had a considerably higher level of procollagen mRNA than the ACL healing site (i.e., injury site) at all postoperative intervals. The results demonstrate that procollagen mRNA levels in MCL tissue are higher than those in ACL tissue under normal conditions, as well as in response to injury. The differences in the procollagen mRNA levels of MCL and ACL may reflect the synthesis of collagen in these tissues, and may contribute to the differences in their healing capacities.

摘要

本研究的目的是比较兔模型中正常和愈合中的内侧副韧带(MCL)及前交叉韧带(ACL)中I型前胶原信使核糖核酸(mRNA)的水平。我们的损伤方法采用了一种手术模型,在MCL和ACL的中间部分造成相同的部分撕裂伤。通过原位杂交研究正常韧带以及撕裂伤后3天、7天、14天和28天的韧带石蜡切片,以比较和追踪两条韧带中I型前胶原mRNA的水平。与α1(I)前胶原mRNA对应的互补DNA(cDNA)探针用[32P]d-CTP进行标记。杂交、放射自显影和切片染色后,通过显微镜检查评估前胶原mRNA的水平。正常MCL中始终检测到比正常ACL更高水平的前胶原mRNA,表明MCL中胶原合成活性更高。在MCL和ACL的损伤部位,I型前胶原mRNA水平在所有撕裂伤后的时期均升高,在术后14天达到最高水平。在所有术后时间段,MCL愈合部位的前胶原mRNA水平均显著高于ACL愈合部位(即损伤部位)。结果表明,在正常条件下以及对损伤的反应中,MCL组织中的前胶原mRNA水平高于ACL组织。MCL和ACL前胶原mRNA水平的差异可能反映了这些组织中胶原的合成情况,并可能导致它们愈合能力的差异。

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