berberine chloride mediates its anti-leishmanial activity via differential regulation of the mitogen activated protein kinase pathway in macrophages小檗碱氯介导anti-leishmanial活动通过微分调节巨噬细胞的有丝分裂原激活蛋白激酶通路.pdfVIP

Berberine Chloride Mediates Its Anti-Leishmanial Activity via Differential Regulation of the Mitogen Activated Protein Kinase Pathway in Macrophages 1 2¤ 1 1 2 1 Piu Saha , Surajit Bhattacharjee , Avijit Sarkar , Alak Manna , Subrata Majumder , Mitali Chatterjee * 1 Department of Pharmacology, Institute of Post Graduate Medical Education and Research, Kolkata, India, 2 Division of Molecular Medicine, Bose Institute, Kolkata, India Abstract Background: A complex interplay between Leishmania and macrophages influences parasite survival and necessitates disruption of signaling molecules, eventually resulting in impairment of macrophage function. In this study, we demonstrate the immunomodulatory activity of Berberine chloride in Leishmania infected macrophages. Principal Findings: The IC50 of Berberine chloride, a quaternary isoquinoline alkaloid was tested in an amastigote macrophage model and its safety index measured by a cell viability assay. It eliminated intracellular amastigotes, the IC50 being 2.8 fold lower than its IC50 in promastigotes (7.10 mM vs. 2.54 mM) and showed a safety index .16. Levels of intracellular and extracellular nitric oxide (NO) as measured by flow cytometry and Griess assay respectively showed that Berberine chloride in Leishmania infected macrophages increased production of NO. Measurement of the mRNA expression of iNOS, IL-12 and IL-10 by RT-PCR along with levels of IL-12p40 and IL-10 by ELISA showed that in infected macrophages, Berberine chloride enhanced expression of iNOS and IL-12p40, concomitant with a downregulation of IL-10. The phosp