Yan Zhenzhong, Ji Lin
Department of Orthopedics, Changzhou No. 7 People's Hospital, No. 288 Yanling East Road, Economic Development Zone, Changzhou, Jiangsu, 213100, China.
Department of Orthopedics, Changzhou Geriatric Hospital Affiliated to Soochow University, No. 288 Yanling East Road, Economic Development Zone, Changzhou, Jiangsu, 213100, China.
Adv Rheumatol. 2024 Dec 18;64(1):88. doi: 10.1186/s42358-024-00427-2.
We investigated role of haematopoietic cell kinase (Hck) in osteoarthritis (OA) and to explore the underlying mechanisms driving its effects. An OA animal model was established and after OA induction, rats received intra-articular injections of lentivirus twice a week for four weeks. Rats were divided into four groups: control (healthy rats without OA), OA model (rats with induced OA), OA + Len-si-NC (OA rats treated with a non-targeting control lentivirus), and OA + Len-si-Hck (OA rats treated with lentivirus targeting Hck). Blood samples were collected, and serum cytokine levels were measured using ELISA. Afterward, the rats were sacrificed for histological analysis and TUNEL assay. In vitro, IL-1β-treated human chondrocytes were transfected with Hck, and the effects on cell viability, apoptosis, ECM degradation, and JAK-STAT3 signaling were assessed. Colivelin, a JAK-STAT3 agonist, was used to confirm the pathway's involvement. Results indicated increased Hck expression in the cartilage tissues of OA rats and in IL-1β-stimulated chondrocytes. Silencing Hck in vivo reduced IL-6 and TNF-α levels, apoptosis, and preserved cartilage structure. In vitro, Hck knockdown in IL-1β-treated chondrocytes resulted in enhanced cell viability, reduced apoptosis, and decreased ECM degradation. Notably, the expression of MMP3 and MMP13 was significantly lowered, while collagen II and aggrecan levels were restored. Additionally, Hck knockdown inhibited JAK-STAT3 activation, which was evident from reduced levels of phosphorylated JAK1 and STAT3. The addition of colivelin reversed these effects, confirming that Hck mediates its effects through the JAK-STAT3 pathway. Overall, our findings indicate that Hck is critical in OA progression by promoting inflammation, apoptosis, and ECM degradation through the JAK-STAT3 signaling pathway activation.
我们研究了造血细胞激酶(Hck)在骨关节炎(OA)中的作用,并探索其发挥作用的潜在机制。建立了OA动物模型,在诱导OA后,大鼠每周接受两次关节腔内注射慢病毒,共四周。大鼠分为四组:对照组(无OA的健康大鼠)、OA模型组(诱导OA的大鼠)、OA + Len-si-NC组(用非靶向对照慢病毒处理的OA大鼠)和OA + Len-si-Hck组(用靶向Hck的慢病毒处理的OA大鼠)。采集血样,采用酶联免疫吸附测定法(ELISA)检测血清细胞因子水平。之后,处死大鼠进行组织学分析和TUNEL检测。在体外,用Hck转染白细胞介素-1β(IL-1β)处理的人软骨细胞,并评估其对细胞活力、凋亡、细胞外基质(ECM)降解和JAK-STAT3信号传导的影响。使用JAK-STAT3激动剂可立维林来确认该信号通路的参与情况。结果表明,OA大鼠软骨组织和IL-1β刺激的软骨细胞中Hck表达增加。体内沉默Hck可降低IL-6和肿瘤坏死因子-α(TNF-α)水平、细胞凋亡,并保留软骨结构。在体外,IL-1β处理的软骨细胞中Hck基因敲低导致细胞活力增强、细胞凋亡减少和ECM降解减少。值得注意的是,基质金属蛋白酶3(MMP3)和基质金属蛋白酶13(MMP13)的表达显著降低,而Ⅱ型胶原蛋白和聚集蛋白聚糖水平得以恢复。此外,Hck基因敲低抑制了JAK-STAT3激活,这从磷酸化JAK1和STAT3水平降低可以明显看出。添加可立维林可逆转这些作用,证实Hck通过JAK-STAT3信号通路介导其作用。总体而言,我们的研究结果表明,Hck通过激活JAK-STAT3信号通路促进炎症、细胞凋亡和ECM降解,在OA进展中起关键作用。