TGF-β_1刺激下损伤的前交叉韧带和内侧副韧带中BMP-1基因的表达  

Differential BMP-1 expression in injured anterior cruciate ligament and medial collateral ligament fibroblasts induced by TGF-β_1

作　　者：尹琳[1] 谢静[1] 蒋稼欢[1] 王春莉[1] 张艳君[1] 许春明[1] KL Paul Sung 

机构地区：[1]重庆大学生物工程学院国家"111计划"基地,重庆400044 [2]美国加州大学圣迭亚哥分校骨科与生物工程系

出　　处：《第三军医大学学报》2012年第7期647-650,共4页Journal of Third Military Medical University

基　　金：高等学校学科创新引智计划(国家"111计划")(B06023);重庆市科技攻关项目(CSTC2008AB529)~~

摘　　要：目的观察在转化生长因子-β1(transforming growth factor beta1,TGF-β1)作用下,损伤的前交叉韧带(anteri-or cruciate ligament,ACL)和内侧副韧带(medial collateral ligament,MCL)中骨形态发生蛋白-1(bone morphogenetic protein-1,BMP-1)基因的表达,找出TGF-β1、BMP-1之间的关系,揭示机械损伤后ACL和MCL细胞中BMP-1的表达差异。方法采用反转录PCR(RT-PCR)和实时荧光定量PCR方法检测1、5、50 ng/ml TGF-β1处理后2 h受损的ACL和MCL细胞中BMP-1的表达以及5 ng/ml TGF-β1作用2、6、12、24 h受损的ACL和MCL细胞中BMP-1的表达;Western blot检测5 ng/mlTGF-β1处理48 h后受损的ACL和MCL细胞中BMP-1的表达。结果受损的ACL和MCL细胞中BMP-1的基因表达比正常状态下偏高,并随着TGF-β1浓度的增大而增高,在MCL中的增高程度比在ACL中高出近1倍(P<0.05);与正常组相比,在5 ng/ml TGF-β1处理24 h后,ACL细胞中BMP-1的表达在24 h达到最高比例(约为6.1倍),而在MCL中12 h达到最高比例(约为9.84倍,P<0.05)。5 ng/ml TGF-β1处理48 h后BMP-1蛋白也明显上调,与无TGF-β1处理的对照组相比,ACL细胞中BMP-1上调2.32倍,MCL细胞中BMP-1上调3.84倍(P<0.05)。结论 TGF-β1刺激BMP-1的变化可能直接影响到细胞外基质中活性赖氨酰氧化酶的表达,对损伤ACL和MCL的修复有极其重要的参考价值和临床意义。Objective To investigate the differential expression of bone morphogenetic protein-1（BMP-1） in injured anterior cruciate ligament（ACL） and medial collateral ligament（MCL） fibroblasts induced by TGF-β1,and to find out the differences between the poorly self-healing ACL and well functionally self-healing MCL fibroblasts.Methods Fibroblasts were primarily cultured from clinical ACL and MCL samples,and then given 12% mechanical stretch injury and treated by 1,5 and 50 ng/ml TGF-β1 for 2 h,or by 5 ng/ml TGF-β1 for 2,6,12 and 24 h respectively at the same time.The expression of BMP-1 in the above treated and untreated fibroblasts were detected by reversed-transcript PCR and real-time quantitative PCR.Western blotting was used to detect the expression of BMP-1 in the injured fibroblasts induced by 5 ng/ml TGF-β1 for 48 h.Results TGF-β1 treatment resulted in an increased mRNA expression of BMP-1 in the injured fibroblasts in a dose-depended manner,especially in the cells from MCL than in those from ACL（about 1 times higher,P0.05）.Compared with the fibroblasts without TGF-β1 treatment,5 ng/ml TGF-β1 treatment for 24 h made the expression of BMP-1 reach the summit in ACL（6.1 folds higher,P0.05） and in MCL（9.84 folds higher,P0.05）.Compared with the control,the protein expression of BMP-1 were elevated to 2.32 and 3.84 folds higher in ACL and MCL respectively after 5 ng/ml TGF-β1 treatment for 48 h（P0.01）.Conclusion TGF-β1 affects the expression of BMP-1 in the injured fibroblasts,and then directly affects activities of lysyl oxiadse in the extracellular matrix,implying its potential significant value and clinical usage in repair of injured ACL.

关 键 词：前交叉韧带 内侧副韧带 骨形态发生蛋白-1 TGF-Β1 

分 类 号：R318.01[医药卫生—生物医学工程] R322.73[医药卫生—基础医学]