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背景:后交叉韧带对人膝关节结构的稳定以及功能的发挥起着重要作用。与内侧副韧带相比,损伤后的后交叉韧带难以很好的自我愈合,甚至会导致半月板撕裂和软骨损伤。为了提高后交叉韧带的愈合能力,这就需要寻找新的途径来再生和修复损伤的后交叉韧带。近年来的研究表明,赖氨酰氧化酶在组织损伤修复过程中起到非常重要的作用,但其在后交叉韧带修复过程中的分子机制研究尚未涉猎。目的:观察与滑膜细胞共培养后交叉韧带成纤维细胞中赖氨酰氧化酶基因的表达。方法:将第4代的后交叉韧带成纤维细胞和滑膜细胞分别种植于6孔板和Tanswell中。实验分为2组,即后交叉韧带成纤维细胞与滑膜细胞共培养组和后交叉韧带成纤维细胞单层培养组。培养6h后,提取总RNA,通过半定量PCR和实时定量PCR检测单层培养组和共培养组中后交叉韧带成纤维细胞中赖氨酰氧化酶基因表达。结果与结论:与单层培养组相比,赖氨酰氧化酶、赖氨酰氧化酶样蛋白1、赖氨酰氧化酶样蛋白2、赖氨酰氧化酶样蛋白3和赖氨酰氧化酶样蛋白4的基因表达在共培养的后交叉韧带细胞中都明显升高,分别增加了1.1,1.4,1.1,1.3,1.1倍(P<0.05)。单层培养组家族成员在单层培养和共培养中表达情况的差异性,说明细胞之间的相互作用会影响后交叉韧带组织的损伤修复,对后交叉韧带损伤修复的机制研究有极其重要的参考价值。
Background: Posterior cruciate ligament plays an important role in the stability of human knee joint and its function. Compared with the medial collateral ligament, the injured posterior cruciate ligament is difficult to self-healing well, and even lead to meniscal tear and cartilage damage. In order to improve the healing ability of the cruciate ligament, it is necessary to find new ways to regenerate and repair the damaged posterior cruciate ligament. In recent years, studies have shown that lysyl oxidase plays an important role in the tissue damage repair process, but its molecular mechanism in the repair of posterior cruciate ligament has not yet been studied. Objective: To observe the expression of lysyl oxidase gene in the cruciate ligament fibroblasts after co-culture with synovial cells. Methods: The fourth generation of posterior cruciate ligament fibroblasts and synovial cells were planted in 6-well plates and Tanswell respectively. The experiment was divided into two groups, that is, the posterior cruciate ligament fibroblasts and synovial cells co-culture group and the posterior cruciate ligament fibroblasts monolayer culture group. After cultured for 6h, the total RNA was extracted and the expression of lysyl oxidase gene in the posterior cruciate ligament fibroblasts in both monolayer and co-culture groups was detected by semi-quantitative PCR and real-time quantitative PCR. RESULTS AND CONCLUSION: Lysyl oxidase, lysyl oxidase-like protein 1, lysyl oxidase-like protein 2, lysyl oxidase-like protein 3, and lysyl oxidase The gene expression of P-4 was significantly increased in the cocultured posterior cruciate ligament cells by 1.1, 1.4, 1.1, 1.3 and 1.1 times (P <0.05), respectively. The differences in the expression of monolayer culture group in monolayer culture and co-culture indicate that the cell-cell interaction may affect the repair of posterior cruciate ligament tissue, and it is extremely important to study the mechanism of posterior cruciate ligament injury repair Reference value.