背景:骨关节炎(OA)是一种退行性疾病,其特征在于炎症和细胞外基质(ECM)降解,会导致严重的个人残疾和社会负担。日本金鱼草。(LJT)是一种具有抗炎和镇痛作用的柳科植物。在传统的东方医学中,LJT通常用于治疗各种疾病,包括骨关节炎和腰背痛。
目的:为了研究抗凋亡,LJT在IL-1β诱导的小鼠软骨细胞中的抗炎和抗衰老特性,并澄清所涉及的潜在机制。此外,该研究还通过建立骨关节炎小鼠模型来检查LJT的作用。最终目标是确定LJT作为抗骨关节炎药物的机制。
方法:在这项研究中,进行分子对接和网络药理学分析以确定LJT作用的潜伏途径和关键靶标。CCK-8试剂盒用于评估LJT对软骨细胞活力的影响。西方印迹,免疫荧光,TUNEL染色试剂盒,和SA-β-gal染色用于验证LJT对软骨细胞的影响。此外,所以,他,免疫组织化学用于评估LJT对小鼠骨关节炎的影响。进行了体外和体内实验以验证LJT在OA中的潜在机制。
结果:网络药理学分析显示AKT1、PTGS2和ESR1是LJT治疗OA的关键候选靶点。分子对接结果表明,AKT1对LJT的主要成分具有较低的结合亲和力。因此,我们选择了STING,PTGS2的上游监管机构,作为我们的调查目标。分子对接显示谷甾醇,福蒙素,豆甾醇和α-Onocerin,LJT的主要组成部分,与STING具有良好的结合活性。体外实验表明LJT抑制IL-1β介导的炎症介质分泌,软骨细胞的凋亡和衰老。结果表明,LJT消除了小鼠不稳定内侧半月板(DMM)引起的软骨退变。机制研究表明,LJT通过抑制STING/NF-κB信号通路,下调NF-κB激活,从而抑制OA的发展。
结论:LJT通过抑制炎症逆转OA的进展,动物模型和软骨细胞的凋亡和衰老。LJT的作用是通过STING/NF-κB途径介导的。
BACKGROUND: Osteoarthritis (OA) is a degenerative disease, its characteristic lies in the inflammation and extracellular matrix (ECM) degradation, can lead to significant personal disability and social burden. Lycopodium japonicum Thunb. (LJT) is a lycopinaceae plant with anti-inflammatory and analgesic effects. In traditional Oriental medicine, LJT is commonly used to treat a variety of conditions, including osteoarthritis and low back pain.
OBJECTIVE: To investigate the anti-apoptotic, anti-inflammatory and anti-senescence properties of LJT in IL-1β-induced mouse chondrocytes, and to clarify the underlying mechanisms involved. In addition, the study also examined the effects of LJT by establishing a mouse model of osteoarthritis. The ultimate goal is to identify the mechanism of LJT as an anti-osteoarthritis agent.
METHODS: In this research, molecular docking and network pharmacology analysis were performed to identify the latent pathways and key targets of LJT action. The CCK-8 kit was used to evaluate LJT\'s effect on chondrocyte viability. Western blotting, Immunofluorescence, TUNEL staining kit, and SA-β-gal staining were employed to verify LJT\'s impact on chondrocytes. Additionally, SO, HE, and Immunohistochemical were utilized to assess LJT\'s effects on osteoarthritis in mice. In vitro and in vivo experiments were performed to verify the potential mechanism of LJT in OA.
RESULTS: Network pharmacology analysis revealed that AKT1, PTGS2, and ESR1 were the key candidate targets for the treatment of OA with LJT. The results of molecular docking indicated that AKT1 exhibited a low binding affinity to the principal constituents of LJT. Hence, we have chosen
STING, an upstream regulator of PTGS2, as our target for investigation. Molecular docking revealed that sitosterol, formononetin, stigmasterol and alpha-Onocerin, the main components of LJT, have good binding activity with
STING. In vitro experiments showed that LJT inhibited IL-1β-mediated secretion of inflammatory mediators, apoptosis and senescence of chondrocytes. The results showed that LJT abolished cartilage degeneration induced by unstable medial meniscus (DMM) in mice. Mechanism research has shown that LJT by inhibiting the
STING/NF-κB signaling pathways, down-regulating the NF-κB activation, so as to inhibit the development of OA.
CONCLUSIONS: LJT reversed the progression of OA by inhibiting inflammation, apoptosis and senescence in animal models and chondrocytes. The effects of LJT are mediated through the
STING/NF-κB pathway.