Homeopathic medicine Cantharis modulates uropathogenic E. coli (UPEC)-induced cystitis in susceptible mice Cidéli de Paula Coelho a,b , Priscilla Dias Motta a , Mariana Petrillo a , Renata de Oliveira Iovine a , Luciane Costa Dalboni a , Fabiana Rodrigues Santana a , Michelle Sanchez Freitas Correia a , Renato Corrêa Viana Casarin c , Vania Maria Carvalho d , Leoni Villano Bonamin a,⇑ a Paulista University (UNIP), Research Center, Av José Maria Whitaker, 290, São Paulo, Brazil b University of Santo Amaro (UNISA), Rua Professor Enéas de Siqueira Neto, 340, São Paulo, Brazil c Faculdade de Odontologia de Piracicaba, Universidade Estadual de Campinas (UNICAMP), Piracicaba, Brazil d Faculty of Veterinary Medicine, University of São Paulo (USP), Av. Prof. Dr. Orlando Marques de Paiva, 87-Cidade Universitária, São Paulo, Brazil article info Article history: Received 22 September 2016 Received in revised form 11 January 2017 Accepted 16 January 2017 Keywords: Escherichia coli Homeopathy UPEC Cytokines Cantharis abstract Objective: This is a random blinded placebo controlled murine experimental model to study the effects of Cantharis 6 CH, a homeopathic medicine, on E coli-induced cystitis. Methods: 24 adult susceptible female BALB/c mice were inoculated with E coli – UPEC O4:K-:H5 by a transurethral catheter. Cantharis 6cH or vehicle (placebo) was offered to mice by free access into the drinking water (1:100), during 24 h after infection. Spleen, bladder and kidneys were processed for quan- titative histopathology after immunohistochemistry, using anti-CD3, CD79, MIF, NK and VEGF antibodies; the cytokines present in the bladder washing fluid were measured using a LUMINEX-Magpix KIT. Mann- Whitney and Fisher exact test were used as statistical analysis. Results: Cantharis 6 CH increased IL12p40, IFN-c and decreased IL10 concentrations in the bladder fluid (p 6 0.05); in the bladder mucosa, it increased the ratio between B and T lymphocytes (31%) and between B lymphocytes and MIF+ macrophages (57%, p 6 0.05). In the pelvis, instead, it decreased the B/T cells ratio (41%, p 6 0.05) and increased the M1/M2 macrophage ratio (42%, p 6 0.05). No differences were seen in the kidney and spleen analysis. Conclusion: The inverted balance of inflammatory cells and cytokines in bladder and pelvis mucosa shows specific local immune modulation induced by Cantharis 6cH. Ó 2017 Elsevier Ltd. All rights reserved. 1. Introduction Escherichia coli is one of the agents commonly involved in diges- tive and extra-intestinal infections, such as those in the urinary tract infection (UTI), being this pathotype called UPEC (uropatho- genic of E. coli) [1]. The infection happens when uropathogens ascend and colonize the lower or upper parts of the urinary tract, originating cystitis and/or pyelonephritis [2]. That is due to the unbalance of the host-parasite relation which involves two simul- taneous factors: the rupture of the hosts’ organism’s defense mech- anisms and the presence of sufficient number of virulent microorganisms capable of adhering, multiplying and persisting in a portion of the urinary tract [3]. Since uropathogens present great resistance to available antibiotics, exploring alternative strategies for managing UTI is a theme of interest [4–7]. Two adhesins participate in uropathogenic processes: type 1 fringes and pili, both involved in bacteria colonization in the uri- nary tract [8–10]. The interaction between E. coli and the hosts’ tis- sue, however, also depends on the immune condition. It is known that B-lymphocytes produce immunoglobulins capable of fixing complement and inducing bacterial lysis; the pro-inflammatory macrophages (M1) are capable of secreting cytokines, that amplify the interaction between lymphocytes and phagocytes, enhancing the presentation of antigens and specificity of the immune response [11]. A recent study developed in vitro showed that the interaction between B-lymphocytes and macrophages can ease the expression of pro-inflammatory cytokines by the later [12]. The development of medications capable of optimizing those interactions and, thus, decreasing the vulnerability of hosts bearers of UPECs to ascendant and systemic infection, would be very valu- able for controlling UTI in susceptible populations. Homeopathy is http://dx.doi.org/10.1016/j.cyto.2017.01.014 1043-4666/Ó 2017 Elsevier Ltd. All rights reserved. ⇑ Corresponding author at: Paulista University, Graduation Program in Environ- mental and Experimental Pathology, Rua Dr. Bacelar, 1212 – 4th Floor, CEP 04026- 002 São Paulo, Brazil. E-mail address: leonibonamin@unip.br (L.V. Bonamin). Cytokine 92 (2017) 103–109 Contents lists available at ScienceDirect Cytokine journal homepage: www.journals.elsevier.com/cytokine