miR-140在膝骨关节炎软骨炎症中的作用及机制研究

A study of the role and mechanism of miR-140 in cartilage inflammation in knee osteoarthritis

目的:探讨miR-140通过调控VEGF信号通路在膝骨关节炎(KOA)软骨炎症中的作用机制。方法:将30只新西兰兔随机分为健康对照组、假手术组和KOA模型组,每组10只。建模后检测实验兔关节液白介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、血管内皮生长因子(VEGF)及软骨组织miR-140表达水平,苏木精-伊红(HE)染色法观察各组软骨组织形态。取KOA模型组兔膝盖软骨组织制备软骨细胞,并转染miR-140 mimics和miR-140 inhibitor转染,分别为miR-140促进转染组和miR-140抑制转染组,设立空白对照组和阴性对照组。qPCR检测转染细胞miR-140和VEGF mRNA表达水平,Western blot法检测转染细胞VEGF的表达水平。结果:KOA模型组IL-1β、TNF-α和VEGF表达水平高于假手术组和健康对照组,且假手术组高于健康对照组(P<0.05)。KOA模型组与健康对照组和假手术组相比,滑膜细胞层次不清晰、排列无序,表面明显肿胀。KOA模型组miR-140表达水平低于健康对照组和假手术组(P<0.05)。与空白组比较,miR-140促进转染组miR-140相对表达量升高(P<0.05),miR-140抑制转染组miR-140相对表达量降低(P<0.05),阴性对照组不变(P>0.05)。与空白组比较,miR-140促进转染组miR-140相对降低(P<0.05),miR-140抑制转染组VEFG mRNA及其蛋白表达升高(P<0.05),阴性对照组不变(P>0.05)。结论:miR-140可能通过调控VEGF信号通路参与KOA软骨炎症的发生过程。

Objective:To investigate the mechanism of miR-140 in cartilage inflammation in knee osteoarthritis(KOA)by regulating the VEGF signaling pathway.Methods:Thirty New Zealand rabbits were randomly divided into a healthy control group,a sham-operated group,and a KOA model group,with 10 rabbits in each group.After modeling,the expression levels of interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),vascular endothelial growth factor(VEGF),and miR-140 in the joint fluid of the rabbits were measured,and the cartilage tissues of each group were observed by hematoxylin-eosin(HE)staining.Chondrocytes were prepared from the knee cartilage tissues of rabbits in KOA model group,then transfected with miR-140 mimics or miR-140 inhibitor,which were the miR-140 enhanced transfection group and miR-140 inhibited transfection group,respectively.A blank control group and a negative control group were also set up.Expression levels of miR-140 and VEGF mRNA in transfected cells were detected using qPCR.Expression level of VEGF in transfected cells was detected by Western blot.Results:The expression levels of IL-1β,TNF-α,and VEGF in the KOA model group were higher than those in the sham-operated group and healthy control group,and those indexes in the sham-operated group were higher than the healthy control group(P<0.05).Compared to the healthy control and sham-operated control groups,the KOA model group had a poorly defined and disorganized synovial cell hierarchy,and a markedly swollen surface.The expression level of miR-140 in the KOA model group was lower than that in the healthy control and sham-operated control groups(P<0.05).Compared with the blank control group,the expression level of VEGF in the miR-140 enhanced transfection group decreased(P<0.05),that in the miR-140 inhibited transfection group increased(P<0.05),and that in the negative control group were not change(P>0.05).Conclusion:miR-140 may be involved in the process of KOA cartilage inflammation by regulating the VEGF signaling pathway.