牛磺熊去氧胆酸调节lncRNA MALAT1改善骨关节炎软骨细胞糖酵解的机制研究OA

Objective:To investigate the therapeutic potential of Tauroursodeoxycholic acid(TUDCA)in alleviating osteoarthritis(OA)cartilage degeneration by targeting the long non-coding RNA MALAT1(metastasis-associated lung adenocarcinoma transcript 1)and regulating chondrocyte glycolytic metabolism.Methods:Primary chondrocytes were isolated from the knee joints of 30 three-week-old SPF-grade C57BL/6 male mice,which were euthanized via cervical dislocation under 5%isoflurane anesthesia.Cells were randomly divided into six experimental groups:Blank control,TUDCA,IL-1β,IL-1β+TUDCA,IL-1β+sh-lncRNA MALAT1(knockdown),and IL-1β+sh-lncRNA MALAT1+TUDCA.An in vitro osteoarthritis model was established by stimulation with IL-1β(10 ng/mL)for 24 hours.For lncRNA MALAT1 knockdown,chondrocytes were transduced with lentiviral vectors carrying shRNA specifically targeting lncRNA MALAT1.After 48 hours of intervention with TUDCA(100 μM),fluorescence in situ hybridization(FISH)was used to detect changes in lncRNA MALAT1 expression in chondrocytes of each group;Real-time PCR was used to analyze the relative expression level of lncRNA MALAT1 in chondrocytes of each group,while Western blot was used to detect the protein levels of glycolytic enzymes(HK2,LDHA,PKM2)and Caspase-3.Results:FISH results showed that compared with the blank control group,the average fluorescence intensity of lncRNA MALAT1 was significantly increased in the IL-1β group;compared with the IL-1β group,the average fluorescence intensity of lncRNA MALAT1 was significantly decreased in the IL-1β+TUDCA group.Real-time PCR revealed that IL-1β+TUDCA treatment significantly reduced lncRNA MALAT1 levels compared with the IL-1β group(P<0.05).Compared with the IL-1β+sh-lncRNA MALAT1 group,the level of lncRNA MALAT1 was increased in the IL-1β+sh-lncRNA MALAT1+TUDCA group(P<0.05).Western blot demonstrated that TUDCA downregulated IL-1β-induced overexpression of HK2,LDHA,PKM2,and Caspase-3(P<0.05).Compared with the IL-1β+sh-lncRNA MALAT1 group,the expression levels of HK2,LDHA,PKM2,and Caspase-3 were further decreased in the IL-1β+sh-lncRNA MALAT1+TUDCA group(P<0.05).Conclusion:TUDCA mitigates OA cartilage degeneration by restoring cellular glycolytic homeostasis in chondrocytes through regulating the expression of lncRNA MALAT1.This study highlights lncRNA MALAT1 as a potential therapeutic target for OA management.