Melittin acupoint injection in attenuating bone erosion in collagen-induced arthritis mice via inhibition of the RANKL/NF-κB signaling pathway
Melittin acupoint injection in attenuating bone erosion in collagen-induced arthritis mice via inhibition of the RANKL/NF-κB signaling pathway
复制标题
蜂毒肽穴位注射通过抑制 RANKL/NF-κB 信号通路减轻胶原诱导的关节炎小鼠骨侵蚀
DOI：
10.21037/atm-22-6583
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发表时间：
2023-01
影响因子：
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通讯作者：
Weiming Chen
中科院分区：
文献类型：
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作者：
Fenfang Liu;Lu Yang;Fen Chen;Jiping Zhang;Le Yang;Fucheng Qiu;Guangen Zhong;Shan Gao;Weizhe Xi;Meilian Lai;Qiting He;Ying Chen;Weiming Chen
研究背景风湿性关节炎（RA）是一种自身免疫性疾病，可导致慢性关节炎。骨侵蚀是类风湿关节炎最严重的病理状态，是导致关节畸形和残疾的主要原因。蜂毒肽穴位注射是治疗类风湿关节炎的有效中药方法。本研究旨在探讨MAI对RA骨侵蚀的影响并阐明其潜在机制。方法建立胶原诱导性关节炎（CIA）小鼠模型。MAI隔日给药1次，共28天。通过体重、关节炎指数（AI）评分、肿胀关节计数（SJC）评分和后爪厚度评估MAI对关节疾病的影响。采用显微CT（micro-CT）拍摄踝关节影像学改变，病理染色观察组织学改变。通过脏器组织学改变、脾脏指数、血清丙氨酸氨基转移酶（ALT）、天门冬氨酸氨基转移酶（AST）、肌酐（Crea）等指标评价MAI的毒性。通过酶联免疫吸附试验（ELISA）确认细胞因子表达水平，以评估CIA小鼠的免疫力。结果MAI能明显改善CIA小鼠的足爪厚度、AI、足关节肿胀数等临床症状（多数P<0.05或<0.01）。组织病理学分析显示，MAI可减轻炎症细胞浸润、滑膜增生、血管翳形成和骨质侵蚀（均P<0.01）。抗酒石酸酸性磷酸酶（TRAP）染色和Micro-CT显示MAI可通过减少破骨细胞的形成而减轻骨侵蚀（P<0.01）。MAI不仅能减轻小鼠的免疫增强作用[高剂量MAI（HM）组P<0.05]，而且对肝、肾无明显毒副作用（P>0.05）。降低血清IL-6、TNF-α水平，升高IL-4、IL-10水平（多数P<0.05或<0.01）。转录组测序结果表明，MAI影响破骨细胞分化途径基因的表达，其作用与核因子κB配体/核因子κ B（RANKL/NF-κB）途径的受体激活因子有关。结论根据我们的研究结果，MAI可以抑制关节炎症并抑制RANKL/NF-κ B介导的破骨细胞分化，从而挽救CIA小鼠的骨侵蚀，这表明MAI可以成为RA的潜在治疗物质。
Background Rheumatoid arthritis (RA) is an autoimmune disease leading to chronic joint inflammation. Bone erosion is the most serious pathological condition of RA and the main cause of joint deformities and disability. Melittin acupoint injection (MAI) is an effective traditional Chinese medicine (TCM) method for RA treatment. This study aimed to investigate the effect of MAI on RA bone erosion and to elucidate the underlying mechanism. Methods A collagen-induced arthritis (CIA) mouse model was established as the experimental subject. MAI was administrated once every other day for 28 days to mice with CIA. The effects of MAI on joint diseases were assessed by body weight, arthritis index (AI) score, swollen joint count (SJC) score, and hind paw thickness. Ankle radiological changes were captured by micro-computed tomography (micro-CT) and histological changes were observed by pathological staining. Organ histological changes, spleen index, alanine aminotransferase (ALT), aspartate aminotransferase (AST), and creatinine (Crea) levels of serum were tested to evaluate the toxicity of MAI. Cytokine expression levels were confirmed by enzyme-linked immunosorbent assay (ELISA) to evaluate the immunity of CIA mice. Results MAI administration markedly improved the clinical signs of CIA in mice, including hind paw thickness, AI, and the number of swollen paw joints (most of them P<0.05 or even <0.01). According to histopathological analysis, MAI ameliorated inflammatory cell infiltration, synovial hyperplasia, pannus formation, and bone erosion (all P<0.01). Micro-CT and tartrate-resistant acid phosphatase (TRAP) staining (P<0.01) also revealed that MAI could relieve bone erosion via reducing the formation of osteoclasts. Not only could MAI relieve the immunological boost [P<0.05 for the high-dose MAI (HM) group], but also it had no liver or kidney side effects (P>0.05). In addition, it decreased the serum levels of interleukin (IL)-6 and tumor necrosis factor-α (TNF-α) and increased the serum levels of IL-4 and IL-10 (the majority of P<0.05 or even <0.01). Transcriptome sequencing results indicated that MAI affected the expression of osteoclast differentiation pathway genes, which was connected with the receptor activator of the nuclear factor κB ligand/nuclear factor kappa B (RANKL/NF-κB) pathway. Conclusions Based on our findings, MAI could suppress joint inflammation and inhibit RANKL/NF-κB-mediated osteoclast differentiation to rescue bone erosion in CIA mice, suggesting that MAI can be a potentially therapeutic substance for RA.