lncRNA HOXA11-AS抑制骨关节炎大鼠软骨细胞凋亡与基质降解的作用研究

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Abstract Objective To investigate the expression of long-nocoding RNA (lncRNA) HOXA11-AS in osteoarthritis (OA) rat chondrocytes and its effect on chondrocyte apoptosis，matrix degradation and nuclear factor κB (Nuclear factor kappa B，NF-κB) .Methods Rats were injected with Freund's complete adjuvant to construct OA rat model，and chondrocytes were isolated and cultured after modeling.Small interfering RNA (Small interfering RNA，siRNA) technology was used to inhibit the expression of HOXA11-AS in chondrocytes；Cell Counting Kit-8 (CCK-8) was used to to detect the viability of chondrocytes after transfection of siRNA at different times.Quantitative Real-time PCR (qRT-PCR) detected the expression of HOXA11-AS and matrix metallo protein-13(MMP-13)a disintesrin and metalloprotease with thrombospondin type 5 motifs(ADAMTS-5)，CollagenⅡ，aggrecan in chondrocytes.Flow cytometry was used to detect chondrocyte apoptosis.Enzyme-linked immuno sorbent assay (ELISA)was used to detect the expression of BCL2-Associated X Protein（Bax）、B-cell lymphoma-2（Bcl-2）、Caspase-3 in chondrocyte supernatant.And Western blot was used to detect the expression of extracellular matrix related proteins MMP-13，ADAMTS-5，CollagenⅡ，aggrecan and anti-nuclear factor kappa B inhibitory protein（IκBα），anti-phosphorylated nuclear factor kappa B inhibitory protein （p-IκBα）、nuclear factor kappa B（NF-κB 65）.Immunofluorescence staining was used to detect the expression of NF-κB in chondrocyte nuclei.Results Compared with the normal group of chondrocytes，HOXA11-AS was highly expressed in chondrocytes of OA rats.Compared with NC siRNA group，the cell viability of HOXA11-AS siRNA group was significantly decreased.Apoptosis rate，proapoptotic factor Baxandthe expression level of Caspase-3 significantly increased，while the expression level of anti-apoptotic factor Bcl-2 was significantly decreased.Meanwhile，compared with NC siRNA group，the protein and gene expression levels of collagen II and aggrecan in HOXA11-AS siRNA group were significantly higher.The expression levels of MMP-13 and ADAMTS-5 genes and proteins were significantly increased，the expression level of IκBα protein was significantly decreased，and the expression levels of p-IκBα and NF-κB p65 were significantly increased.In addition，NF-κB was significant expression in the nucleusin HOXA11-AS siRNA group.Conclusion lncRNA HOXA11-AS can inhibit the apoptosis and matrix degradation of chondrocytes in rats with osteoarthritis and prevent the activation of NF-κB signaling pathway.

Key words： HOXA11-AS chondrocytes；apoptosis matrix degradation NF-κB signaling pathway expression level

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	Qian Liang
	Li Hongjun
	Shi Yuan
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Qian Liang,Li Hongjun,Shi Yuan, et al. Effect of lncRNA HOXA11-AS on the Inhibition of Chondrocyte Apoptosis and Matrix Degradation in Osteoarthritis Rats[J]. sygkzz, 2020, 26(7): 614-623.

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http://www.sygkzz.com/EN/ OR http://www.sygkzz.com/EN/Y2020/V26/I7/614

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