Sirin Duygu Yasar, Karaarslan Numan
Department of Molecular Biology and Genetics, Faculty of Arts and Sciences, Namik Kemal University, Tekirdag, Turkey.
Department of Neurosurgery, School of Medicine, Namik Kemal University, Tekirdag, Turkey.
Arch Med Sci. 2018 Oct;14(6):1340-1347. doi: 10.5114/aoms.2018.73134. Epub 2018 Feb 2.
The aim of the present study is to investigate the effects of pregabalin (PGB) on chondrocyte proliferation and collagen type II (), hypoxia-inducible factor 1-α (α), and chondroadherin () gene expression in osteoarthritic chondrocytes.
Standard primary chondrocyte cultures were prepared using osteochondral tissues that were surgically obtained from 6 patients with gonarthrosis. Cell morphology was evaluated using an inverted microscope, and cell death and proliferation were determined through MTT analysis, which was confirmed by AO/PI staining and statistically evaluated. The expression levels of , , and α genes were assessed using gene-specific TaqMan Gene Expression Assays.
MTT analyses showed that PGB administration did not have a negative or toxic effect on cell viability and proliferation in cultured chondrocytes ( < 0.001), but in our morphological evaluation extracellular matrix development was observed to be weaker in cultures treated with PGB. After 24 h of treatment, COL2A1, HIF-1α, and CHAD gene expression decreased in the groups to which PGB was applied compared to gene expression before the experiment (at 0 h); at 48 h, CHAD and HIF-1α expression increased to the same level as the control group, but the expression of COL2A1 continued to decrease.
Further studies need to be conducted with more participants to prove that there is a negative correlation between extracellular matrix formation and PGB administration. Our preliminary data show that even at low doses and over short-term administration, PGB may affect chondrocyte cells at the gene-expression level.
本研究旨在探讨普瑞巴林(PGB)对骨关节炎软骨细胞中软骨细胞增殖、Ⅱ型胶原蛋白(COL2A1)、缺氧诱导因子1-α(HIF-1α)和软骨黏附蛋白(CHAD)基因表达的影响。
使用从6例膝关节病患者手术获取的骨软骨组织制备标准原代软骨细胞培养物。使用倒置显微镜评估细胞形态,并通过MTT分析确定细胞死亡和增殖情况,AO/PI染色对此进行了确认并进行统计学评估。使用基因特异性TaqMan基因表达分析方法评估COL2A1、CHAD和HIF-1α基因的表达水平。
MTT分析表明,给予PGB对培养的软骨细胞的细胞活力和增殖没有负面或毒性作用(P<0.001),但在我们的形态学评估中,观察到用PGB处理的培养物中细胞外基质的发育较弱。处理24小时后,与实验前(0小时)的基因表达相比,应用PGB的组中COL2A1、HIF-1α和CHAD基因表达下降;在48小时时,CHAD和HIF-1α表达增加至与对照组相同水平,但COL2A1的表达继续下降。
需要更多参与者进行进一步研究,以证明细胞外基质形成与给予PGB之间存在负相关。我们的初步数据表明,即使在低剂量和短期给药的情况下,PGB也可能在基因表达水平上影响软骨细胞。