BaiLab时讯：青蒿素通过抑制Wntβ-Catenin信号通路缓解骨关节炎
Cell Physiol Biochem.  2018 Dec   doi: 10.1159/000495926.
Background/Aims: proliferation
Current drug therapies for osteoarthritis (OA) are not practical because of the cytotoxicity and severe side-effects associated with most of them. Artemisinin (ART), an antimalarial agent, is well known for its safety and selectivity to kill injured cells. Based on its anti-inflammatory activity and role in the inhibition of OA-associated Wnt/β-catenin signaling pathway, which is crucial in the pathogenesis of OA, we hypothesized that ART might have an effect on OA.
Methods: 
The chondro-protective and antiarthritic effects of ART on interleukin-1-beta (IL-1β)-induced and OA patient-derived chondrocytes were investigated in vitro using cell viability assay, glycosaminoglycan secretion, immunofluorescence, quantitative reverse transcription-poly
merase chain reaction, and western blotting. We also used OA model rats constructed by anterior cruciate ligament transection and medial meniscus resection (ACLT+MMx) in the joints to investigate the effects of ART on OA by gross observation,morphological staining, immunohistochemistry, and enzyme-linked immunosorbent assay.
Results: 
ART exhibited potent anti-inflammatory effects by inhibiting the expression of proinflammatory chemokines and cytokines, including interleukin (IL)-1β, IL-6, tumor necrosis factor alpha, and matrix metallopeptidase-13. It also showed favorable chondro-protective effect as evidenced by enhanced cell proliferation and viability, increased glycosaminoglycan deposition, prevention of chondrocyte apoptosis, and degeneration of cartilage. Further, ART inhibited OA progression and cartilage degradation via the Wnt/β-catenin signaling pathway, suggesting that it might serve as a Wnt/β-catenin antagonist to reduce inflammation and prevent cartilage degradation. 
Conclusion: 
In conclusion, ART alleviates IL-1β-mediated inflammatory response and OA progression by regulating the Wnt/β-catenin signaling pathway. Thereby, it might be developed as a potential therapeutic agent for OA.
摘要：
研究背景及目的：目前用于骨关节炎（OA）的众多药物由于其细胞毒性以及严重的副作用在临床上使用受限，我们知道青蒿素（ART）是一种抗疟药，因为能够选择性杀伤受损细胞而闻名，最近研究发现其具有很强的抗炎活性以及抑制Wnt /β-catenin信号通路功能，基于此作者提出假设，青蒿素对骨关节炎的病理进展可能具有一定作用。方法：体外实验主要是通过分离大鼠以及人原代关节软骨细胞，探讨青蒿素对于白细胞介素-1-β（IL-1β）刺激后关节软骨细胞代谢的响体。体外实验则主要通过切断前交叉韧带和切除关节内侧半月板（ACLT + MMx）构建的OA模型大鼠，通过大体观察，形态学染，免疫组织化学研究青蒿素对OA病理进展的影响。结果:青蒿素通过抑制促炎因子的表达表现出有效的抗炎作用；通过抑制软骨细胞凋亡从而促进其增值，同时抑制软骨细胞外基质的降解。青蒿素发挥以上功能主要是通过抑制Wnt /β-catenin信号传导途径。结论：ART通过调节Wnt /β-catenin信号通路从而来延缓OA进展。
Fig1. ART对IL-1β诱导的大鼠软骨细胞具有保护作用
（a）首先通过MTT试验选取用于后续细胞实验所用青蒿素的最佳浓度。发现0.125–4.0 μg/mL这个浓度范围可明显促进大鼠软骨细胞增值，因此选择浓度为4.0 μg/mL作为后续实验。（b）、（d）通过MTT以及FDA / PI染检测其对细胞增殖以及细胞活性的影响。（c）通过番红O染检测其对软骨细胞合成GAG（蛋白聚糖）的影响。（e）、（f）、（g）分别通过qRT-PCR、WB、IF检测ART对炎性因子（IL-1β、IL-6、TNF-α）以及细胞凋亡指标（BAX、caspase-3）、细胞外基质代谢（MMP-13）的影响。
Fig 2.体外，ART对于人体关节软骨细胞具有保护功能。
分离OA病人以及正常人关节软骨细胞原代培养，于第二天、第四天以及第六天进行检测。实验结果与大鼠原代软骨细胞结果一致。（a）、（b）通过MTT以及FDA / PI染发现ART可以逆转OA关节软骨细胞的凋亡。（c）、（d）ART可明显促进软骨细胞合成蛋白聚糖。（e）、（f）、（g）分别通过qRT-PCR、WB、IF证实了ART可明显抑制OA关节软骨细胞中炎性因子（IL-1β、IL-6、TNF-α）、凋亡聚蛋白(BAX、caspase-3)以及基质金属蛋白酶(MMP-13)的合成。

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