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Chlorogenic acid acts upon Leishmania donovani arresting cell cycle and modulating cytokines and nitric oxide in vitro.
Parasite Immunology ( IF 1.4 ) Pub Date : 2020-04-20 , DOI: 10.1111/pim.12719
Nilanjana Majumder 1 , Subhrajit Ganguly 1 , Arijit Kumar Ghosh 1 , Shreetoma Kundu 1 , Antara Banerjee 2 , Samiran Saha 1
Affiliation  

AIMS Visceral leishmaniasis (VL), caused by Leishmania donovani in India, is fatal if untreated, having serious concern of limited chemotherapeutic options. In this study, we evaluated antileishmanial efficacy of purified chlorogenic acid (CGA) against promastigotes and intracellular amastigotes infected into RAW264.7 macrophages. METHODS AND RESULTS Chlorogenic acid was effective both on promastigotes (IC50 = 78.394 µmol/L, i.e. 27.75 µg/mL) and intracellular amastigotes (ED50 = 26.752 µmol/L, i.e. 9.47 µg/mL). In promastigotes, significant retardation in mitotic growth was caused both by cell-death and reduction of metabolic activity, evidenced by propidium-iodide uptake and MTT assay, respectively. Flow cytometric analysis revealed that retardation of mitotic growth was due to cell-cycle arrest at G1/S checkpoint. Complete clearance of amastigotes from infected RAW264.7 cells, assessed by microscopic counting, was achieved with 60 µmol/L (21.24 µg/mL) CGA for 24 hours, with negligible toxicity to host macrophages. This parasite clearing efficacy was comparable to 1.0 µg/mL (1.082 µmol/L) Amphotericin B, and 20 µmol/L Miltefosine, two standard antileishmanial drugs. Cytokine-ELISA revealed that elevated IL-10 production by infected macrophages was reduced after parasite clearance. Consequently, IL-12, TNF and NO (assayed by Griess test) production by macrophages were significantly increased after successful resolution of infection. CONCLUSION Chlorogenic acid might emerge as a potential antileishmanial drug.

中文翻译:

绿原酸可在体外抑制多形性利什曼原虫的细胞周期并调节细胞因子和一氧化氮。

AIMS由印度利什曼原虫引起的内脏利什曼病(VL),如果不加以治疗,将是致命的,严重关注有限的化学疗法选择。在这项研究中,我们评估了纯化的绿原酸(CGA)对被RAW264.7巨噬细胞感染的前鞭毛体和胞内变形虫的抗疟疾功效。方法和结果绿原酸对前鞭毛体(IC50 = 78.394 µmol / L,即27.75 µg / mL)和胞内变形虫(ED50 = 26.752μmol/ L,即9.47 µg / mL)均有效。在前鞭毛体中,细胞死亡和代谢活性的降低均引起有丝分裂生长的显着延迟,分别由碘化丙锭摄取和MTT测定法证明。流式细胞仪分析表明,有丝分裂生长的延迟是由于细胞周期停滞在G1 / S检查点。通过显微镜计数评估,用60 µmol / L(21.24 µg / mL)CGA进行24小时的发酵,可以完全清除受感染的RAW264.7细胞中的变形虫,对宿主巨噬细胞的毒性可忽略不计。这种寄生虫清除功效可与两种标准的抗盲肠药物安非他命A(两性霉素B)和安非他命(20 mg / L)两性霉素B的浓度相媲美,为1.0 µg / mL(1.082 µmol / L)。细胞因子酶联免疫吸附测定显示,寄生虫清除后,感染的巨噬细胞产生的高水平的IL-10减少。因此,在成功解决感染后,巨噬细胞的IL-12,TNF和NO的产量(通过Griess试验测定)显着增加。结论绿原酸可能是一种潜在的抗衰老药物。对宿主巨噬细胞的毒性可忽略不计。这种寄生虫清除功效可与两种标准的抗盲肠药物安非他命(Amphotericin B)和1.0 µg / mL(1.082 µmol / L)两性霉素B和20 µmol / L Miltefosine媲美。细胞因子酶联免疫吸附测定表明,寄生虫清除后,感染的巨噬细胞产生的高水平的IL-10减少。因此,在成功解决感染后,巨噬细胞的IL-12,TNF和NO的产量(通过Griess试验测定)显着增加。结论绿原酸可能是一种潜在的抗衰老药物。对宿主巨噬细胞的毒性可忽略不计。这种寄生虫清除功效可与两种标准的抗盲肠药物安非他命A(两性霉素B)和安非他命(20 mg / L)两性霉素B的浓度相媲美,为1.0 µg / mL(1.082 µmol / L)。细胞因子酶联免疫吸附测定显示,寄生虫清除后,感染的巨噬细胞产生的高水平的IL-10减少。因此,在成功解决感染后,巨噬细胞的IL-12,TNF和NO的产量(通过Griess试验测定)显着增加。结论绿原酸可能是一种潜在的抗衰老药物。成功解决感染后,巨噬细胞产生的IL-12,TNF和NO(通过Griess试验测定)显着增加。结论绿原酸可能是一种潜在的抗衰老药物。成功解决感染后,巨噬细胞产生的IL-12,TNF和NO(通过Griess试验测定)显着增加。结论绿原酸可能会成为一种潜在的抗衰老药物。
更新日期:2020-04-05
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