Department of Physical Medicine and Rehabilitation, Chang Gung Memorial Hospital, Taoyuan City 33302, Taiwan.
Graduate Institute of Clinical Medical Sciences, Chang Gung University, Taoyuan City 33302, Taiwan.
Int J Mol Sci. 2022 Aug 7;23(15):8787. doi: 10.3390/ijms23158787.
Lidocaine injection is a common treatment for tendon injuries. However, the evidence suggests that lidocaine is toxic to tendon cells. This study investigated the effects of lidocaine on cultured tendon cells, focusing on the molecular mechanisms underlying cell proliferation and extracellular matrix (ECM) production. Tendon cells cultured from rat Achilles tendons were treated with 0.5, 1.0, or 1.5 mg/mL lidocaine for 24 h. Cell proliferation was evaluated by Cell Counting Kit 8 (CCK-8) assay and bromodeoxyuridine (BrdU) assay. Cell apoptosis was assessed by Annexin V and propidium iodide (PI) stain. Cell cycle progression and cell mitosis were assessed through flow cytometry and immunofluorescence staining, respectively. The expression of cyclin E, cyclin A, cyclin-dependent kinase 2 (CDK2), p21, p27, p53, matrix metalloproteinases-2 (MMP-2), matrix metalloproteinases-9 (MMP-9), type I collagen, and type III collagen were examined through Western blotting, and the enzymatic activity of MMP-9 was determined through gelatin zymography. Lidocaine reduced cell proliferation and reduced G1/S transition and cell mitosis. Lidocaine did not have a significant negative effect on cell apoptosis. Lidocaine significantly inhibited cyclin A and CDK2 expression but promoted p21, p27, and p53 expression. Furthermore, the expression of MMP-2 and MMP-9 increased, whereas that of type I and type III collagen decreased. Lidocaine also increased the enzymatic activity of MMP-9. Our findings support the premise that lidocaine inhibits tendon cell proliferation by changing the expression of cell-cycle-related proteins and reduces ECM production by altering levels of MMPs and collagens.
利多卡因注射是治疗肌腱损伤的常用方法。然而，有证据表明利多卡因对肌腱细胞有毒性。本研究探讨了利多卡因对培养的肌腱细胞的影响，重点研究细胞增殖和细胞外基质（ECM）产生的分子机制。从大鼠跟腱中培养的肌腱细胞用 0.5、1.0 或 1.5mg/ml 利多卡因处理 24 小时。通过细胞计数试剂盒 8（CCK-8）和溴脱氧尿苷（BrdU）检测评估细胞增殖。通过 Annexin V 和碘化丙啶（PI）染色评估细胞凋亡。通过流式细胞术和免疫荧光染色分别评估细胞周期进程和细胞有丝分裂。通过 Western blot 检测细胞周期蛋白 E、细胞周期蛋白 A、细胞周期蛋白依赖性激酶 2（CDK2）、p21、p27、p53、基质金属蛋白酶-2（MMP-2）、基质金属蛋白酶-9（MMP-9）、I 型胶原和 III 型胶原的表达，并通过明胶酶谱法测定 MMP-9 的酶活性。利多卡因降低细胞增殖，降低 G1/S 期转变和细胞有丝分裂。利多卡因对细胞凋亡没有显著的负面影响。利多卡因显著抑制细胞周期蛋白 A 和 CDK2 的表达，但促进 p21、p27 和 p53 的表达。此外，MMP-2 和 MMP-9 的表达增加，而 I 型和 III 型胶原的表达减少。利多卡因还增加了 MMP-9 的酶活性。我们的研究结果支持这样一个前提，即利多卡因通过改变细胞周期相关蛋白的表达抑制肌腱细胞增殖，并通过改变 MMP 和胶原的水平减少 ECM 的产生。
