Ma Cong, Wen Bo, Zhang Qin, Shao Pei-Pei, Gu Wen, Qu Kun, Shi Yang, Wang Bei
Department of Rheumatology and Immunology, Beijing Hospital of Traditional Chinese Medicine, Capital Medical University, Beijing 100010, People's Republic of China,
Drug Des Devel Ther. 2019 Feb 11;13:601-609. doi: 10.2147/DDDT.S182087. eCollection 2019.
Ankylosing spondylitis (AS) is a type of rheumatoid disease, which has been reported to be associated with the excessive proliferation of fibroblasts recently. Emodin, a single component from a traditional Chinese medicine , exerts anti-inflammation and antirheumatic arthritis activities. However, could emodin be used to treat AS remains unclear? Thus, this study aimed to investigate the effect of emodin on AS.
Fibroblasts obtained from patients with AS were used in the current study. In addition, multiple cellular and molecular biology techniques such as Cell Counting Kit-8, Western blotting, flow cytometry, monodansylcadaverine staining, and immunofluorescence assay were applied as well.
Emodin-induced apoptosis of fibroblasts obtained from patient with AS via increasing active caspase-9, active caspase-3, and Bax levels and downregulating Bcl-2. Meanwhile, emodin enhanced autophagy in fibroblasts via upregulation of the expression of Atg12, Atg5, and Beclin 1, which was further confirmed by monodansylcadaverine staining. As expected, autophagy inhibitor 3-methyladenine (3MA) completely reversed emodin-induced autophagy in fibroblasts. Moreover, 3MA significantly increased emodin-induced apoptosis of fibroblasts obtained from patient with AS by increasing the levels of γH2AX, active caspase-9, active caspase-3, and cleaved poly ADP-ribose polymerase.
Our results indicated that emodin effectively induced apoptosis and autophagy of fibroblasts obtained from patient with AS. In addition, suppression of autophagy enhanced emodin-induced apoptosis in fibroblasts. Therefore, we proposed that combination of emodin with autophagy inhibitor might be a potent strategy for improving the symptoms of AS in the future.
强直性脊柱炎（AS）是一种类风湿性疾病，最近有报道称其与成纤维细胞过度增殖有关。大黄素是一种来自中药的单一成分，具有抗炎和抗风湿性关节炎的活性。然而，大黄素是否可用于治疗AS仍不清楚。因此，本研究旨在探讨大黄素对AS的影响。
本研究使用从AS患者获取的成纤维细胞。此外，还应用了多种细胞和分子生物学技术，如细胞计数试剂盒-8、蛋白质印迹法、流式细胞术、单丹磺酰尸胺染色和免疫荧光测定法。
大黄素通过增加活化的半胱天冬酶-9、活化的半胱天冬酶-3和Bax水平以及下调Bcl-2诱导AS患者来源的成纤维细胞凋亡。同时，大黄素通过上调Atg12、Atg5和Beclin 1的表达增强成纤维细胞中的自噬，单丹磺酰尸胺染色进一步证实了这一点。正如预期的那样，自噬抑制剂3-甲基腺嘌呤（3MA）完全逆转了大黄素诱导的成纤维细胞自噬。此外，3MA通过增加γH2AX、活化的半胱天冬酶-9、活化的半胱天冬酶-3和切割的聚ADP-核糖聚合酶的水平，显著增加了大黄素诱导的AS患者来源的成纤维细胞凋亡。
我们的结果表明，大黄素有效地诱导了AS患者来源的成纤维细胞凋亡和自噬。此外，自噬的抑制增强了大黄素诱导的成纤维细胞凋亡。因此，我们提出大黄素与自噬抑制剂联合使用可能是未来改善AS症状的有效策略。
