北京中医药大学学报2025，Vol.48Issue(1)：55-67,13.DOI:10.3969/j.issn.1006-2157.2025.01.008
升麻素苷介导lncRNA NEAT1/miR-128-3p改善骨关节炎软骨细胞胆固醇代谢的机制研究
The mechanism of Prim-O-glucosylcimifugin in improving cholesterol metabolism in osteoarthritis chondrocytes via lncRNA NEAT1/miR-128-3p
摘要
Abstract
Objective To investigate the mechanism of action of Prim-O-glucosylcimifugin(POG)to improve cholesterol metabolism in osteoarthritic(OA)chondrocytes based on the long noncoding RNA nuclear-enriched transcript 1(lncRNA NEAT1)/microRNA-128-3p(miR-128-3p)pathway.Methods For in vivo experiments,60 mice were divided into the normal,sham operation,model,and POG groups using the random number table method,with 15 mice per group.The osteoarthritis mouse model was constructed using the modified Hulth method in the model and POG groups.Mice in the POG group were administered 30 mg/(kg·d)POG by gavage.The other groups were administered an equal amount of normal saline for 8 weeks.The cartilage tissue structure of mice in each group was observed using hematoxylin and eosin staining.Real-time PCR was used to detect changes in the lncRNA NEAT1 and miR-128-3p mRNA expression levels in the cartilage tissues of mice.Western blotting was used to detect the protein expressions of ATP-binding cassette transporter A1(ABCA1),liver X receptor β(LXRβ),matrix metalloprotein-3(MMP-3),and B-lymphoblastoma-2-associated X protein(Bax)in articular cartilage of mice.An enzyme-linked immunosorbent assay was used to measure the tumor necrosis factor-α(TNF-α)content in the synovial fluid of mice.A biochemical microplate assay was used to measure the total cholesterol level in the synovial fluid of mice.The in vitro experiments were divided into the negative control,interleukin-1β(IL-1β),IL-1β+POG,IL-1β+oe-lncRNA NEAT1,IL-1β+oe-lncRNA NEAT1+POG,IL-1β+miR-128-3p inhibition,and IL-1β+miR-128-3p inhibition+POG groups.An OA model was established by inducing chondrocytes with IL-1β for 24 h,and 90 mg/L of POG and miR-128-3p inhibitor(50 nmol/L)were administered for 48 h as an intervention.lncRNA NEAT1 expression in chondrocytes was detected using fluorescence in situ hybridization.A dual luciferase assay was used to detect the targeting relationship between lncRNA NEAT1 and miR-128-3p.Lentiviral plasmids overexpressing lncRNA NEAT1 were used to transfect mouse chondrocytes.Real-time PCR was used to detect the effect of lncRNA NEAT1 overexpression on the mRNA level of miR-128-3p in chondrocytes.Western blotting was used to detect ABCA1,LXRβ,MMP-3,and Bax protein expression in chondrocytes after lncRNA NEAT1 overexpression and miR-128-3p inhibition.Results POG significantly reduced OA cartilage tissue damage.Compared with the model group,the lncRNA NEAT1 mRNA level decreased,whereas the miR-128-3p mRNA level increased in the cartilage tissue of the POG group(P<0.05).Compared with the model group,ABCA1 and LXRβ protein expression increased in the POG group,whereas MMP-3 and Bax protein expression decreased(P<0.05).The TNF-α levels decreased in the POG group compared to the model group(P<0.05).Compared with the model group,the total cholesterol level in the synovial fluid of the joint of mice in the POG group decreased(P<0.05).The mean fluorescence intensity of lncRNA NEAT1 in the IL-1β+POG group decreased compared with the IL-1β group(P<0.05).The relative luciferase activity in the miR-128-3p mimics group bound to the lncRNA NEAT1-WT plasmid decreased compared with the miR-128-3p negative control group(P<0.05).The lncRNA NEAT1 mRNA levels decreased,whereas the miR-128-3p mRNA levels increased in the IL-1β+oe-lncRNA NEAT1+POG group compared with the IL-1β+oe-lncRNA NEAT1 group(P<0.05).Compared with the IL-1β+POG group,ABCA1 and LXRβ protein expression decreased,whereas MMP-3 and Bax protein expression increased(P<0.05).Conclusion POG mediates lncRNA NEAT1/miR-128-3p to improve cholesterol metabolism in OA chondrocytes.关键词
升麻素苷/骨关节炎/软骨细胞/胆固醇代谢/小鼠Key words
Prim-O-glucosylcimifugin/osteoarthritis/chondrocytes/cholesterol metabolism/mice分类
医药卫生引用本文复制引用
林艳铭,涂海水,兰书洁,李超,陆诗雨,陈悦,付长龙..升麻素苷介导lncRNA NEAT1/miR-128-3p改善骨关节炎软骨细胞胆固醇代谢的机制研究[J].北京中医药大学学报,2025,48(1):55-67,13.基金项目
国家自然科学基金项目(No.82104888) （No.82104888）
陈可冀中西医结合发展基金项目(No.CKJ2023004) （No.CKJ2023004）
福建中医药大学校管项目(No.X2023024) （No.X2023024）
福建中医药大学高层次人才科研启动资金项目(No.X2019011-人才) National Natural Science Foundation of China(No.82104888) （No.X2019011-人才）
