Evaluation of the antibacterial, anti-inflammatory, anti-osteoclastogenic and anti-HIV activities of Malva sylvestris

Author(s)

Bruna Benso

• Document type: Doctoral Thesis
• Press: Piracicaba, SP.
• Institution: Universidade Estadual de Campinas (UNICAMP). Faculdade de Odontologia de Piracicaba
• Defense date: 2016-02-18

Examining board members

Pedro Luiz Rosalen; Severino Matias de Alencar; Carina Denny; Janaina de Cassia Orlandi Sardi; Francisco Carlos Groppo

• Advisor: Gilson Cesar Nobre Franco; Pedro Luiz Rosalen

Abstract

Nature has been a source of medicinal products for centuries, yielding many useful drugs. A wide variety of plants are well recognized for their medicinal and nutraceutical value, Malva sylvestris being one example; the ethnopharmacological literature has reported a long history of recognition of biological properties. The aim of this study was to conduct a pharmacological screening of Malva sylvestris and its interest. Therefore, we investigated: (1) the antimicrobial and anti-inflammatory activity of M. sylvestris extract (MSE) and fractions using a cell culture technique with epithelial and gingival cells infected with Aggregatibacter actinomycetemcomitans and a gene expression and cytokine quantification related to the inflammatory response; (2) The activity of MSE and fractions in the in vivo anti-inflammatory activity (neutrophil migration, paw edema and cytokine quantification, anti-osteoclastogenic action (gene expression, number of positive TRAP positive cells and zymography), antioxidant activity (DPPH and ABTS¿+), and chemical identification of the bioactive fraction (MS/MS); (3) anti-HIV activity of aqueous fraction (AF) in cells infected with HIV-Bal using the in vitro dual chamber model, quantifying p24 antigen, gene expression and cytokines. Statistical analysis were performed by analysis of variance (ANOVA) and Dunnett¿s post-hoc test. The results showed that chloroform fraction CLF (75 ?g/mL) was efficient in reducing the bacteria colonization and inflammatory mediators, promoting the gene regulation of IL-1beta, IL-6, IL-10, CD14, PTGS, MMP-1 and FOS, as well as reducing protein expression IL-1beta, IL-6, IL-8 and GM-CSF. The in vivo reduction of anti-inflammatory effect (30 mg/kg, orally) was significant for the extract (MSE) and all fractions (CLF, EAF and AF) with the exception of the hexane fraction in the neutrophil migration assay. The AF (30 mg/kg, orally) reduced the paw edema in the first 3 hours analyzed, with a faster action than the positive control, reducing the levels of IL-1? expression. The activity of M. sylvestris in the bone remodeling assay demonstrated that the aqueous fraction (AF) in the concentration of 10 ?g/mL regulated the gene transcription of the study genes (carbonic anhydrase, cathepsin K and tartrate-resistant acid phosphatase) and reduced the number of TRAP-positive osteoclasts and the specific proteolytic enzyme MMP-9. In terms of the antioxidant activity, the AF and the ethyl acetate fraction (EAF) had the best ability to capture free radicals. The chemical identification revealed rutin as the bioactive compound in the AF. Results for the antiviral activity showed a p24 antigen reduction, reverse transcriptase mechanism of action, controlled transcription of the genes CD4, Bcl-2 and TRIM5, and a reduction in the cytokines IL-beta, IL-6, IL-8 and GM-CSF after treatment with AF (50 ?g/mL). Therefore, we can conclude that M. sylvestris and its bioactive fractions are promising compounds as novel therapeutic agents (AU)

• FAPESP's process: 11/23980-5 - Evaluation of the potential antimicrobial, atiinflamatory, bone remodeling, antioxidant and anticancer of Malva sylvestris.
• Grantee: Bruna Benso
• Support Opportunities: Scholarships in Brazil - Doctorate