摘要
目的探究lncRNA HAND2-AS1对膝骨关节炎软骨退变的影响及其机制。方法SD大鼠随机分为假手术组(Sham组)、膝骨关节炎组(KOA组)、膝骨关节炎+过表达对照组(KOA+ov-NC组)和膝骨关节炎+过表达HAND2-AS1组(KOA+ov-HAND2-AS1组),每组10只,观察大鼠形态变化。RT-qPCR检测各组大鼠关节软骨中HAND2-AS1、miR-449a和RNF125 mRNA表达水平;HE染色观察各组大鼠关节软骨病理损伤;TUNEL实验检测各组大鼠关节软骨细胞凋亡情况;Western blot检测各组大鼠关节软骨中RNF125、β-catenin、p-GSK3β和p-β-catenin蛋白表达水平。生物信息学预测miR-449a与HAND2-AS1和RNF125的潜在结合位点,并采用双荧光素酶报告基因实验验证。结果过表达lncRNA HAND2-AS1改善膝骨关节炎大鼠关节炎症状和关节软骨病理损伤,抑制大鼠关节软骨细胞凋亡,下调大鼠关节软骨中miR-449a、β-catenin和p-GSK3β表达,并上调RNF125和p-β-catenin表达。miR-449a与HAND2-AS1和RNF125 mRNA结合。结论过表达lncRNA HAND2-AS1改善膝骨关节炎诱导的软骨退变,其机制与海绵化miR-449a上调RNF125表达并抑制Wnt/β-catenin信号通路激活有关。
Objective To explore the effect and its mechanism of lncRNA HAND2-AS1 on cartilage degeneration of knee osteoarthritis.Methods SD rats were randomly divided into sham group(Sham group),knee osteoarthritis group(KOA group),knee osteoarthritis+overexpression control group(KOA+ov-NC group)and knee osteoarthritis+overexpression HAND2-AS1 group(KOA+ov-HAND2-AS1 group),with 10 rats in each group.The morphological changes of rats were observed.RT-qPCR was used to detect the levels of HAND2-AS1 mRNA,miR-449a and RNF125 mRNA in articular cartilage of rats in each group.HE staining was used to observe the pathological damage of articular cartilage in rats in each group.TUNEL was used to detect the apoptosis in articular chondrocytes of rats in each group.Western blot was used to detect the expressions of RNF125,βcatenin,p-GSK3β and p-β-catenin in the articular cartilage of rats in each group.Bioinformatics predicted the potential binding sites of miR-449a to HAND2-AS1 and RNF125,and verified them by dual luciferase reporter gene assay.Results Overexpression of lncRNA HAND2-AS1 could improve the arthritis symptoms and pathological damage of articular cartilage in rats with knee osteoarthritis,inhibit the apoptosis of cells in articular cartilage of rats,down-regulate the expressions of miR 449a,β-catenin and p-GSK3βin articular cartilage of rats and up regulate the expressions of RNF125 and p-β-catenin.miR-449a binds to HAND2-AS1 and RNF125 mRNA.Conclusion Overexpression of lncRNA HAND2-AS1 improves cartilage degeneration induced by knee osteoarthritis,and its mechanism is related to sponge miR-449a up-regulating RNF125 expression and inhibiting Wntβcatenin signal pathway activation.
作者
陈星宇
任奋强
周斌
艾力亚尔·阿依都
王新安
CHEN Xing-yu;REN Fen-qiang;ZHOU Bin;ELIAR Ayidu;WANG Xin-an(Department of Trauma Repair Surgery,Bazhou People's Hospital,Korla 841099,China)
出处
《解剖科学进展》
2025年第5期651-654,659,共5页
Progress of Anatomical Sciences
基金
新疆医科大学研究生创新创业项目(CXCY2023012)。
