Mohanty Soumitra, Kamolvit Witchuda, Zambrana Silvia, Sandström Corine, Gonzales Eduardo, Östenson Claes-Göran, Brauner Annelie
Department of Microbiology, Tumour and Cell Biology, Division of Clinical Microbiology, Karolinska Institutet and Karolinska University Hospital, 17176 Stockholm, Sweden.
Department 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.
J Ethnopharmacol. 2017 Feb 23;198:214-220. doi: 10.1016/j.jep.2017.01.011. Epub 2017 Jan 10.
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.
The extract was analyzed 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.
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.
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.
玻利维亚风轮菜是一种具有抗炎和抗感染活性的南美植物。抗生素耐药性的不断增加促使人们寻求替代疗法。基于其在传统医学中的应用，我们研究了玻利维亚风轮菜对膀胱上皮细胞抵御大肠杆菌感染能力的影响。
采用液相色谱 - 质谱联用仪对提取物进行分析。使用膀胱上皮细胞系T24和5637、尿路致病性大肠杆菌12号菌株、其同基因突变体WE16 csgBA bscA::Cm以及CFT073来研究玻利维亚风轮菜对尿道上皮感染的影响。分析了玻利维亚风轮菜处理后的细胞上细菌的黏附和侵袭情况。采用逆转录 - 聚合酶链反应评估了玻利维亚风轮菜处理后尿血小板溶素1a、β1整合素、小窝蛋白 - 1、白细胞介素 - 8和抗菌肽的表达。通过蛋白质印迹分析或酶联免疫吸附测定法确认蛋白质表达。使用最低抑菌浓度研究了玻利维亚风轮菜对细菌和真菌的抗菌作用。此外，用结晶紫测定法研究了生物膜的形成。
玻利维亚风轮菜提取物由70多种不同类型的植物化学物质组成，包括糖类和酚类化合物。该提取物降低了尿血小板溶素1a的表达以及大肠杆菌对尿道上皮细胞的黏附和侵袭，同时上调了小窝蛋白 - 1的表达。在未感染且经玻利维亚风轮菜处理的细胞中，白细胞介素 - 8的水平低于未处理的细胞。然而，在感染细胞中，处理组和未处理组之间未观察到差异。此外，玻利维亚风轮菜处理可减少尿路致病性大肠杆菌（UPEC）生物膜的形成，但不抑制细菌生长。
我们的结果表明，玻利维亚风轮菜通过减少细菌黏附、侵袭和生物膜形成，对膀胱上皮细胞抵御UPEC感染具有保护作用。
