Transcriptional analysis and target genes discovery of Pseudomonas aeruginosa biofilm developed ex vivo chronic wound model

AMB Express

Xiaojuan Tan¹  Xi Cheng¹  Yifan Zhang¹  Guoping Zhu¹  Mei Hu¹  Jinwei Zhou²  Aiqun Jia³ 

[1] Anhui Provincial Key Laboratory of Molecular Enzymology and Mechanism of Major Diseases, Anhui Normal University, 241000, Wuhu, Anhui, China;Key Laboratory of Biomedicine in Gene Diseases and Health of Anhui Higher Education Institutes, College of Life Sciences, Anhui Normal University, 241000, Wuhu, Anhui, China;School of Food and Biology Engineering, Xuzhou University of Technology, 221018, Xuzhou, China;School of Life and Pharmaceutical Sciences, State Key Laboratory of Marine Resource Utilization in South China Sea, Hainan University, 570228, Haikou, China;

关键词: Pseudomonas aeruginosa;    Biofilms;    Chronic wound infections;    Alkaline phosphatase;    Nitric oxide;   

DOI  :  10.1186/s13568-021-01317-2

来源: Springer

【 摘 要 】

Bacterial biofilms formation is one of the major reasons for treatment failure in chronic wound infections. Therefore, diagnostic biomarkers remain the best option for prevention and treatment of chronic wound infections by biofilms. Herein, Pseudomonas aeruginosa PAO1 was used to mimic biofilm development in porcine skin explants wells as ex vivo wound model. The microscopic imaging showed that PAO1 in porcine skin explants wells formed micro-colonies at 24 h, developed mushroom-like structure at 48 h, and at 72 h mushroom-like structure disappeared, remaining a thin bacterial lawn. RNA-seq data analysis revealed that the expression levels of genes involved in the type II hxc secretion system were significantly higher in biofilms than in planktonic cells, especially the expression of lapA encoding alkaline phosphatase. However, the expression levels of genes associated with denitrification pathway were markedly decreased in biofilms, especially the transcription of nirS encoding nitrite reductase to produce nitric oxide (NO). Therefore, their expressions and products were further detected using RT-qPCR and biochemical assays, respectively. The results found that the expression of lapA and alkaline phosphatase activity were induced, but the expression of nirS and intracellular NO were reduced at the whole biofilms cycle. The study indicates that LapA and NO would play an important role for P. aeruginosa biofilm formation in chronic wound infections. LapA would serve as potential target to monitor chronic wound infections by P. aeruginosa biofilms. Inducing NO would be used to treat chronic wound infections due to P. aeruginosa biofilms.

【 授权许可】

CC BY