Zhang Kun, Gao Shan, Guo JiaYi, Ni GuoHua, Chen Zhe, Li Feng, Zhu XiaoLei, Wen YongBing, Guo YanXing
Department of Orthopedics, No. 91 Central hospital of Liberation Army, Jiaozuo 454150, Henan province, China.
Faculty of Graduate Studies, Hunan University of Chinese Medicine, Changsha 410208, Hunan Province, China.
Iran J Basic Med Sci. 2018 Feb;21(2):130-137. doi: 10.22038/IJBMS.2018.23871.5991.
To elucidate the effects and potential mechanisms of hypericin-photodynamic therapy (HYP-PDT) for treating the human rheumatoid arthritis (RA) fibroblast-like synoviocyte (FLS) MH7A cell-line.
MH7A cells were subjected to HYP-PDT intervention and apoptosis was evaluated via MTT, nuclear staining, and flowcytometry analyses. Intracellular reactive oxygen species (ROS) were measured with the fluorescent probe 2'7'-dichlorofluorescein diacetate (DCFH-DA). To verify the effects of HYP on apoptotic and nuclear factor kappa-B (NF-κB) pathways, caspase-8, 9, poly-ADP-ribose polymerase (PARP), phosphorylated (p)-NF-κB p65, NF-κB p65 and p-IκBα protein expressions were quantified with Western blot. Quantitative real-time PCR was used to assay NF-κB p65 mRNA.
HYP-PDT inhibited MH7A cell viability and induced apoptosis in a dose-dependent manner. Meanwhile, intracellular ROS levels increased significantly after HYP-PDT treatment. Furthermore, the expression of cleaved caspase-9 and PARP was increased by HYP-PDT treatment, with a concurrent decline in NF-κB.
HYP-PDT induces apoptosis in MH7A cells, at least partially, via generation of ROS, regulation of the apoptotic pathway and suppression of the NF-κB pathway. These findings suggest that HYP-PDT may be a potential treatment for RA.
阐明金丝桃素光动力疗法(HYP-PDT)治疗人类风湿关节炎(RA)成纤维样滑膜细胞(FLS)MH7A细胞系的效果及潜在机制。
对MH7A细胞进行HYP-PDT干预,并通过MTT法、核染色和流式细胞术分析评估细胞凋亡情况。用荧光探针2',7'-二氯二氢荧光素二乙酸酯(DCFH-DA)检测细胞内活性氧(ROS)水平。为验证金丝桃素对凋亡和核因子κB(NF-κB)通路的影响,采用蛋白质印迹法对caspase-8、9、聚ADP核糖聚合酶(PARP)、磷酸化(p)-NF-κB p65、NF-κB p65和p-IκBα蛋白表达进行定量分析。采用定量实时PCR检测NF-κB p65 mRNA。
HYP-PDT以剂量依赖方式抑制MH7A细胞活力并诱导细胞凋亡。同时,HYP-PDT处理后细胞内ROS水平显著升高。此外,HYP-PDT处理使裂解的caspase-9和PARP表达增加,同时NF-κB表达下降。
HYP-PDT至少部分通过产生活性氧、调节凋亡通路和抑制NF-κB通路诱导MH7A细胞凋亡。这些研究结果表明,HYP-PDT可能是一种潜在的RA治疗方法。