Department of Pathology, School of Basic Medicine, Anhui Medical University, Hefei, Anhui Province, China.
School of Pharmacy, Anhui Medical University, Hefei, Anhui Province, China.
Pharmacol Rep. 2019 Dec;71(6):1006-1013. doi: 10.1016/j.pharep.2019.05.014. Epub 2019 May 22.
Approaches promoting fibroblast-like synoviocytes (FLS) apoptosis are considered as a meaningful strategy for rheumatoid arthritis (RA) treatment. We have previously reported the anti-arthritic effect of penta-acetyl geniposide ((Ac)GP, an active derivative of geniposide) on adjuvant-induced arthritis (AIA) rats in vivo. The present study aimed to investigate the pro-apoptotic effect of (Ac)GP on AIA FLS in vitro and the underlying molecular mechanisms.
Rat AIA was induced by complete Freund's adjuvant, and FLS were primary-cultured from synovial tissues. AIA FLS were treated with (Ac)GP (50, 100 and 200 μM) for 48 h and cell proliferation and apoptosis were respectively examined. The involvement of apoptosis-related proteins (Bax, Bcl-2 and caspase 3) and nuclear factor kappa B (NF-κB) signaling pathway was checked.
(Ac)GP inhibited the viability of AIA FLS and reduced the percentage of Ki67-positive cells in AIA FLS. Particularly, (Ac)GP promoted AIA FLS apoptosis in vitro by inducing apoptotic nuclear morphology, facilitating DNA ladder formation and increasing percentages of both early and late apoptotic cells. (Ac)GP treatment on AIA FLS decreased Bcl-2 protein level whereas increased the levels of Bax and caspase 3 proteins. Moreover, (Ac)GP reduced the degradation and phosphorylation of IκBα, down-regulated NF-κB p65 protein level in nucleus and inhibited NF-κB p65 nuclear translocation.
(Ac)GP had a potent pro-apoptotic effect on AIA FLS in vitro, which is associated with regulating apoptosis-related proteins and inhibiting NF-κB activation.
促进成纤维样滑膜细胞（FLS）凋亡的方法被认为是治疗类风湿关节炎（RA）的一种有意义的策略。我们之前已经报道过 penta-acetyl geniposide（（Ac）GP，geniposide 的一种活性衍生物）在体内对佐剂诱导的关节炎（AIA）大鼠的抗关节炎作用。本研究旨在探讨（Ac）GP 在体外对 AIA FLS 的促凋亡作用及其潜在的分子机制。
用完全弗氏佐剂诱导大鼠 AIA，并从滑膜组织中原代培养 FLS。用（Ac）GP（50、100 和 200μM）处理 AIA FLS 48 小时，分别检测细胞增殖和凋亡。检查凋亡相关蛋白（Bax、Bcl-2 和 caspase 3）和核因子 kappa B（NF-κB）信号通路的参与情况。
（Ac）GP 抑制 AIA FLS 的活力，减少 AIA FLS 中 Ki67 阳性细胞的百分比。特别是，（Ac）GP 通过诱导凋亡核形态、促进 DNA 梯形成以及增加早期和晚期凋亡细胞的比例，在体外促进 AIA FLS 凋亡。（Ac）GP 处理 AIA FLS 降低了 Bcl-2 蛋白水平，而增加了 Bax 和 caspase 3 蛋白水平。此外，（Ac）GP 减少了 IκBα的降解和磷酸化，降低了核内 NF-κB p65 蛋白水平，并抑制了 NF-κB p65 的核转位。
（Ac）GP 在体外对 AIA FLS 具有很强的促凋亡作用，这与调节凋亡相关蛋白和抑制 NF-κB 激活有关。
