Extract of Clinopodium bolivianum protects against E. coli invasion of uroepithelial cells.

J Ethnopharmacol. 2017 Jan 10. pii: S0378-8741(16)31228-4. doi: 10.1016/j.jep.2017.01.011. [Epub ahead of print]

Mohanty S¹, Kamolvit W¹, Zambrana S², Sandström C³, Gonzales E⁴, Östenson CG⁵, Braunerf A⁶.

Author information

• ¹Department of Microbiology, Tumour and Cell Biology, Division of Clinical Microbiology, Karolinska Institutet and Karolinska University Hospital, 17176 Stockholm, Sweden.
• ²Department of Molecular Medicine and Surgery, Karolinska Institutet and Karolinska University Hospital, 17176 Stockholm, Sweden; Area de Farmacologia, Instituto de Investigaciones Farmaco Bioquimicas, Facultad de Ciencias Farmacéuticas y Bioquimicas, Universidad Mayor de San Andrés, La Paz, Bolivia.
• ³Department of Chemistry and Biotechnology, Uppsala BioCenter, Swedish University of Agricultural Sciences, 750 07 Uppsala, Sweden.
• ⁴Area de Farmacologia, Instituto de Investigaciones Farmaco Bioquimicas, Facultad de Ciencias Farmacéuticas y Bioquimicas, Universidad Mayor de San Andrés, La Paz, Bolivia.
• ⁵Department of Molecular Medicine and Surgery, Karolinska Institutet and Karolinska University Hospital, 17176 Stockholm, Sweden.
• ⁶Department of Microbiology, Tumour and Cell Biology, Division of Clinical Microbiology, Karolinska Institutet and Karolinska University Hospital, 17176 Stockholm, Sweden. Electronic address: Annelie.Brauner@ki.se.

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE:

Clinopodium bolivianum is a South American plant with anti-inflammatory and anti-infective activities. The increasing antibiotic resistance urges for alternative therapy. Based on its use in traditional medicine, we investigated the effect of C. bolivianum on the ability to defend bladder epithelial cells from E. coli infection.

MATERIALS AND METHODS:

The extract was analysed by LC-MS. Bladder epithelial cell lines T24 and 5637 and uropathogenic E. coli No. 12, its isogenic mutant WE16 csgBA bscA::Cm and CFT073 were used to investigate the effect of C. bolivianum on uroepithelial infection. Bacterial adherence and invasion to cells treated with C. bolivianum were analyzed. Expression of uroplakin 1a, β1 integrin, caveolin-1, IL-8 and antimicrobial peptides in response to C. bolivianum treatment was assessed using RT-PCR. Protein expression was confirmed by Western blot analysis or ELISA. The antimicrobial effects of C. bolivianum on bacteria and fungus were investigated using minimum inhibitory concentration. Furthermore, the formation of biofilm was investigated with crystal violet assay.

RESULTS:

C. bolivianum extract consisted of more than 70 different types of phytochemicals including sugars and phenolic compounds. The extract decreased the uroplakin 1a expression and E. coli adhesion and invasion of uroepithelial cells while up-regulated caveolin-1. In uninfected C. bolivianum treated cells, IL-8 was lower than in non-treated cells. In infected cells, however, no difference was observed between treated and non-treated cells. Further, C. bolivianum treatment reduced uropathogenic E. coli (UPEC) biofilms but did not inhibit bacterial growth.

CONCLUSIONS:

Our results show that C. bolivianum has a protective role on bladder epithelial cells against UPEC infection by decreasing the bacterial adhesion, invasion and biofilm formation.

Copyright © 2017 Elsevier Ireland Ltd. All rights reserved.

KEYWORDS:

Clinopodium bolivianum; E. coli; biofilm; urinary tract infection; uroepithelial cells