Hashiguchi Motoko, Suzuki Kaori, Kaneko Kazuo, Nagaoka Isao
Department of Medicine for Motor Organ, Juntendo University Graduate School of Medicine, Tokyo 113‑8421, Japan.
Department of Host Defense and Biochemical Research, Juntendo University Graduate School of Medicine, Tokyo 113‑8421, Japan.
Mol Med Rep. 2017 Jun;15(6):4398-4404. doi: 10.3892/mmr.2017.6541. Epub 2017 May 2.
Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by synovial hyperplasia. Methotrexate (MTX), an antifolate derivative, is used for the treatment of RA, as it exerts antiproliferative efftects on lymphocytes and synovial cells. Aloe‑emodin (AE) is a primary component of anthraquinones in Aloe vera and exerts antiproliferative and apoptotic effects on various tumor cells. In the present study, the effect of AE on the proliferation and apoptosis of MH7A human RA synovial cells was examined. In addition, the effect of AE was compared with that of the established RA therapeutic MTX. MH7A cells were incubated with 5, 10, 20 or 40 µM AE, or 0.01, 0.05, 0.1 or 1 µM MTX, for 24, 48 or 72 h. Subsequently, total cell numbers were assessed using trypan blue staining and Cell Counting kit‑8. Furthermore, MH7A cells incubated with AE or MTX for 48 h were evaluated for apoptosis following Annexin V/propidium iodide (PI) staining, and for cell cycle distribution following PI staining. The results indicated that ≥10 µM AE and ≥0.05 µM MTX effectively decreased the numbers of viable MH7A cells. In addition, 40 µM AE and 1 µM MTX induced apoptosis in MH7A cells. Cell cycle analysis revealed that ≥20 µM AE induced G2/M phase arrest, whereas ≥0.1 µM MTX induced S phase arrest. These observations suggested that AE treatment inhibited the growth of MH7A cells by arresting the cell cycle at a different checkpoint compared with MTX treatment. Thus, AE may be a potential therapeutic agent for the treatment of RA, and may be complimentary to MTX, based on its antiproliferative effect on synovial cells.
类风湿关节炎(RA)是一种以滑膜增生为特征的慢性炎症性疾病。甲氨蝶呤(MTX)是一种抗叶酸衍生物,因其对淋巴细胞和滑膜细胞具有抗增殖作用而用于治疗RA。芦荟大黄素(AE)是芦荟中蒽醌类的主要成分,对多种肿瘤细胞具有抗增殖和凋亡作用。在本研究中,检测了AE对MH7A人RA滑膜细胞增殖和凋亡的影响。此外,将AE的作用与已确立的RA治疗药物MTX的作用进行了比较。将MH7A细胞与5、10、20或40 μM AE,或0.01、0.05、0.1或1 μM MTX孵育24、48或72小时。随后,使用台盼蓝染色和细胞计数试剂盒-8评估总细胞数。此外,对用AE或MTX孵育48小时的MH7A细胞进行膜联蛋白V/碘化丙啶(PI)染色后评估凋亡情况,并用PI染色评估细胞周期分布。结果表明,≥10 μM AE和≥0.05 μM MTX有效降低了存活的MH7A细胞数量。此外,40 μM AE和1 μM MTX诱导MH7A细胞凋亡。细胞周期分析显示,≥20 μM AE诱导G2/M期阻滞,而≥0.1 μM MTX诱导S期阻滞。这些观察结果表明,与MTX治疗相比,AE治疗通过在不同的检查点阻滞细胞周期来抑制MH7A细胞的生长。因此,基于其对滑膜细胞的抗增殖作用,AE可能是治疗RA的潜在治疗药物,并且可能与MTX互补。