长链非编码RNA RP11在膝关节骨性关节炎软骨细胞损伤的作用及机制

The Role and Mechanism of Long Chain Non Coding RNA RP11 in Articular Cartilage Injury of Knee Osteoarthritis

为了探讨长链非编码RNA(lncRNA)RP11对膝关节骨性关节炎(OA)软骨细胞的影响,本研究选取膝关节OA软骨组织和正常软骨组织,分离软骨细胞;同时将OA软骨组织细胞随机分为对照组、空白转染组和RP11 shRNA组,其中空白转染组转染空载质粒脂质体,RP-11 shRNA组转染RP-11 shRNA。采用PCR检测RP11 mRNA表达,CCK8细胞增殖活力实验检测细胞活力,流式细胞仪检测细胞凋亡,Western blotting检测MMP-3、MMP-9和MMP-13蛋白表达。结果表明,OA软骨细胞RP11mRNA相对表达量为(0.683±0.095),明显高于正常软骨细胞(p<0.05);OA软骨细胞增殖活力为(42.01±10.30)%,明显低于正常软骨细胞(p<0.05),而细胞凋亡率为(59.82±8.78)%,明显高于正常软骨细胞(p<0.05);RP11-shRNA组RP11 mRNA相对表达量和细胞凋亡率分别为(0.322±0.100)和(44.82±9.23)%,明显低于对照组和空白转染组(p<0.05),而细胞增殖活力为(52.11±8.34)%,明显高于对照组和空白转染组(p<0.05);RP11-shRNA组MMP-3、MMP-9和MMP-13蛋白相对表达量分别为(0.503±0.098)、(0.202±0.082)和(0.335±0.098),明显低于对照组和空白转染组(p<0.05)。lncRNA RP11可影响膝关节OA软骨细胞增殖及凋亡,可能与调控MMP-3、MMP-9和MMP-13蛋白有关。

To investigate the effect of long chain non coding RNA(lncRNA)RP11 on chondrocytes of knee osteoarthritis(OA).Chondrocyteswere isolated fromOAcartilage tissue and normal cartilage tissue of knee joint;OA chondrocytes were randomly divided into control group,blank transfection group and RP11 shRNA group,the blank transfection group was transfected with empty plasmid liposomes,and RP-11 shRNA group transfected RP-11 shRNA.RP11 mRNA expression was detected by PCR,cell viability was detected by CCK8 cell proliferation assay,apoptosis was detected by flow cytometry,and the expressions of MMP-3,MMP-9 and MMP-13 were detected by Western blotting.The relative expression of RP11 mRNA in OA chondrocytes was(0.683±0.095),which was significantly higher than that in normal chondrocytes(p<0.05);The proliferative activity of OA chondrocytes was(42.01±10.30)%,which was significantly lower than that of normal chondrocytes(p<0.05),the apoptosis rate was(59.82±8.78)%,which was significantly higher than that of normal chondrocytes(p<0.05);The relative expression of RP11 mRNA and the apoptosis rate of cells in RP11-shRNA group were(0.322±0.100)and(44.82±9.23)%,which were significantly lower than those in control group and blank transfection group(p<0.05),while the proliferation activity of cells was(52.11±8.34)%,which were significantly higher than those in control group and blank transfection group(p<0.05);The relative expression levels of MMP-3,MMP-9 and MMP-13 protein in RP11-shRNA group were(0.503±0.098),(0.202±0.082)and(0.335±0.098),were significantly lower than those in control group and blank transfection group(p<0.05).LncRNA RP11 can affect the proliferation and apoptosis of OA chondrocytes in knee joint,which may be related to the regulation ofMMP-3,MMP-9 andMMP-13 proteins.