Wang Hailong, Jiang Quan, Feng Xinghua
J Tradit Chin Med. 2017 Apr;37(2):207-13. doi: 10.1016/s0254-6272(17)30046-8.
To investigate the effects of icariin on apoptosis and the expression of Fas, Fas ligand (FasL), B cell lymphoma (Bcl-2), and Bcl-2-associated X protein (Bax) in CD4+ T lymphocytes from patients with ankylosing spondylitis.
Primary cultures of peripheral blood CD4+ T lymphocytes were established and treated with icariin at high, medium, and low doses (0.5, 0.25, and 0.125 mg/mL). Sulfasalazine treated and helthy cells were used as controls. Apoptosis of treated cells was determined by flow cytometry. Reverse transcription polymerase chain reaction and enzyme-linked immunosorbent assays were used to determine the effects of icariin on the expression of Fas, FasL, Bcl-2, and Bax. The activity of caspase 8 and caspase 3 was determined by a colorimetric assay.
The mRNA and protein expression of Fas, and activity of caspase 8 and caspase 3 in CD4+ T lymphocytes were increased by icariin (P < 0.05). Conversely, the mRNA and protein expression of Bcl-2 was decreased (P < 0.05). The expression of FasL and Bax were not significantly different between groups. The proapoptotic effects of icariin were dose-dependent.
Icariin induces the apoptosis of CD4 + T cells from patients with AS comparing to normal control. Therefore, the induction of apoptosis may be the likely mechanism of action of icariin's antirheumatics activities.
探讨淫羊藿苷对强直性脊柱炎患者CD4+T淋巴细胞凋亡及Fas、Fas配体(FasL)、B细胞淋巴瘤-2(Bcl-2)和Bcl-2相关X蛋白(Bax)表达的影响。
建立外周血CD4+T淋巴细胞原代培养物,并用高、中、低剂量(0.5、0.25和0.125mg/mL)的淫羊藿苷进行处理。以柳氮磺胺吡啶处理的细胞和健康细胞作为对照。通过流式细胞术测定处理后细胞的凋亡情况。采用逆转录聚合酶链反应和酶联免疫吸附测定法来确定淫羊藿苷对Fas、FasL、Bcl-2和Bax表达的影响。通过比色法测定半胱天冬酶8和半胱天冬酶3的活性。
淫羊藿苷可增加CD4+T淋巴细胞中Fas的mRNA和蛋白表达以及半胱天冬酶8和半胱天冬酶3的活性(P<0.05)。相反,Bcl-2的mRNA和蛋白表达降低(P<0.05)。各组之间FasL和Bax的表达无显著差异。淫羊藿苷的促凋亡作用呈剂量依赖性。
与正常对照相比,淫羊藿苷可诱导强直性脊柱炎患者CD4 + T细胞凋亡。因此,诱导凋亡可能是淫羊藿苷抗风湿活性的作用机制。