Abstract：
BACKGROUND: Facet joint osteoarthritis (FJOA) is a prevalent condition contributing to low back pain, particularly in the elderly population. This study aimed to investigate the potential role of Cytokine Receptor-like Factor 1 (CRLF1) in FJOA pathogenesis and its therapeutic implications.
METHODS: Bioinformatics analysis was utilized to identify CRLF1 as the target gene, followed by quantification of CRLF1 expression levels and joint degeneration degree using immunohistochemistry (IHC). In primary chondrocytes, the inhibition of CRLF1 expression by siRNA was performed, and Western blot analysis was conducted to evaluate the involvement of the extracellular matrix and MAPK/ERK signaling pathway. Flow cytometry was employed to assess the apoptosis rate of chondrocytes, while immunofluorescence (IF) was utilized to evaluate the localization of CRLF1, cleaved-caspase3, MMP13, COL2A1, and ERK.
RESULTS: The expression of CRLF1 was found to be significantly elevated in FJOA tissues compared to normal tissues. Through the use of loss-of-function assays, it was determined that CRLF1 not only enhanced the rate of apoptosis in chondrocytes, but also facilitated the degradation of the extracellular matrix in vitro. Furthermore, CRLF1 was found to activate the ERK1/2 pathways. The pro-arthritic effects elicited by CRLF1 were mitigated by treatment with the MEK inhibitor U0126 in chondrocytes.
CONCLUSIONS: These results suggest that CRLF1 enhances chondrocytes apoptosis and extracellular matrix degration in FJOA and thus may therefore be a potential therapeutic target for FJOA.
METHODS: Bioinformatics analysis was utilized to identify CRLF1 as the target gene, followed by quantification of CRLF1 expression levels and joint degeneration degree using immunohistochemistry (IHC). In primary chondrocytes, the inhibition of CRLF1 expression by siRNA was performed, and Western blot analysis was conducted to evaluate the involvement of the extracellular matrix and MAPK/ERK signaling pathway. Flow cytometry was employed to assess the apoptosis rate of chondrocytes, while immunofluorescence (IF) was utilized to evaluate the localization of CRLF1, cleaved-caspase3, MMP13, COL2A1, and ERK.
RESULTS: The expression of CRLF1 was found to be significantly elevated in FJOA tissues compared to normal tissues. Through the use of loss-of-function assays, it was determined that CRLF1 not only enhanced the rate of apoptosis in chondrocytes, but also facilitated the degradation of the extracellular matrix in vitro. Furthermore, CRLF1 was found to activate the ERK1/2 pathways. The pro-arthritic effects elicited by CRLF1 were mitigated by treatment with the MEK inhibitor U0126 in chondrocytes.
CONCLUSIONS: These results suggest that CRLF1 enhances chondrocytes apoptosis and extracellular matrix degration in FJOA and thus may therefore be a potential therapeutic target for FJOA.
摘要：
背景：小关节骨关节炎（FJOA）是导致下背痛的常见病，尤其是老年人。本研究旨在探讨细胞因子受体样因子1（CRLF1）在FJOA发病机制中的潜在作用及其治疗意义。
方法：利用生物信息学分析鉴定CRLF1为靶基因，随后使用免疫组织化学(IHC)定量CRLF1表达水平和关节变性程度。在原代软骨细胞中，通过siRNA进行CRLF1表达的抑制，并进行Westernblot分析以评估细胞外基质和MAPK/ERK信号通路的参与。流式细胞术用于评估软骨细胞的凋亡率，而免疫荧光（IF）用于评估CRLF1，裂解的caspase3，MMP13，COL2A1和ERK的定位。
结果:发现与正常组织相比,CRLF1在FJOA组织中的表达显著升高。通过使用功能丧失测定法，确定CRLF1不仅提高了软骨细胞的凋亡率，而且在体外也促进了细胞外基质的降解。此外，发现CRLF1激活ERK1/2途径。通过用MEK抑制剂U0126在软骨细胞中处理,减轻了由CRLF1引起的促关节炎作用。
结论：这些结果表明，CRLF1可增强FJOA的软骨细胞凋亡和细胞外基质降解，因此可能是FJOA的潜在治疗靶点。
方法：利用生物信息学分析鉴定CRLF1为靶基因，随后使用免疫组织化学(IHC)定量CRLF1表达水平和关节变性程度。在原代软骨细胞中，通过siRNA进行CRLF1表达的抑制，并进行Westernblot分析以评估细胞外基质和MAPK/ERK信号通路的参与。流式细胞术用于评估软骨细胞的凋亡率，而免疫荧光（IF）用于评估CRLF1，裂解的caspase3，MMP13，COL2A1和ERK的定位。
结果:发现与正常组织相比,CRLF1在FJOA组织中的表达显著升高。通过使用功能丧失测定法，确定CRLF1不仅提高了软骨细胞的凋亡率，而且在体外也促进了细胞外基质的降解。此外，发现CRLF1激活ERK1/2途径。通过用MEK抑制剂U0126在软骨细胞中处理,减轻了由CRLF1引起的促关节炎作用。
结论：这些结果表明，CRLF1可增强FJOA的软骨细胞凋亡和细胞外基质降解，因此可能是FJOA的潜在治疗靶点。
