Qingre Baidu mixture‑induced effect of AI‑2 on Staphylococcus aureus and Pseudomonas aeruginosa biofilms in chronic and refractory wounds

Shan,Wei; Wang,Yunfei; Zhang,Zhen; Xing,Jie; Xu,Jienan; Xiao,Wen; Shen,Yiting; Guo,Shuyu; Que,Huafa

doi:10.3892/etm.2019.7391

Qingre Baidu mixture‑induced effect of AI‑2 on Staphylococcus aureus and Pseudomonas aeruginosa biofilms in chronic and refractory wounds

Authors:
- Wei Shan
- Yunfei Wang
- Zhen Zhang
- Jie Xing
- Jienan Xu
- Wen Xiao
- Yiting Shen
- Shuyu Guo
- Huafa Que
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Affiliations: Department of Surgery of Traditional Chinese Medicine, Longhua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 200032, P.R. China
Published online on: March 13, 2019 https://doi.org/10.3892/etm.2019.7391
Pages: 3343-3350
Copyright: © Shan et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The present study aimed to investigate the effect of the traditional Chinese medicine Qingre Baidu mixture (QBM) on the regulation of various parameters, including the morphology of bacterial biofilms, the bacterial density sensing system, the self‑induction of the molecule autoinducer (AI)‑2 and the hypoxia‑inducible factor (HIF)‑vascular endothelial growth factor (VEGF) signaling pathway. For that purpose, Sprague Dawley rats were administered the QBM, the Wu Wei Xiao Du Wan drink (WXD) and cefoperazone (Cef) prior to drug isolation from serum. Vibrio harveyi BB170 was employed as a reporter strain to detect the AI‑2 signaling pathway in Staphylococcus aureus and Pseudomonas aeruginosa. The expression of HIF‑1α and VEGF expression was examined by immunohistochemistry. ELISAs were used to measure the expression of HIF‑1α, HIF‑2α, HIF‑3α and VEGF. The level of inflammation was evaluated by hematoxylin and eosin staining. Biofilm formation and the number of macrophages were detected by immunofluorescence. The results revealed that the QBM could reduce the concentration of AI‑2 derived from Staphylococcus aureus and Pseudomonas aeruginosa, and markedly increase the levels of HIF‑1α, HIF‑2α and VEGF in chronic and refractory wounds. The QBM strongly inhibited the formation of bacterial biofilms and the number of macrophages, therefore promoting wound healing. In conclusion, the QBM could inhibit the biofilm formation of Staphylococcus aureus and Pseudomonas aeruginosa through decreasing the levels of AI‑2 while upregulating the expression of HIF‑1α, HIF‑2α and HIF‑3α, which increased the levels of VEGF, thereby promoting angiogenesis and wound healing in chronic and refractory wounds.

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1	Merckoll P, Jonassen TØ, Vad ME, Jeansson SL and Melby KK: Bacteria, biofilm and honey: A study of the effects of honey on ‘planktonic’ and biofilm-embedded chronic wound bacteria. Scand J Infect Dis. 41:341–347. 2009. View Article : Google Scholar : PubMed/NCBI
2	Clinton A and Carter T: Chronic wound biofilms: Pathogenesis and potential therapies. Lab Med. 46:277–284. 2015. View Article : Google Scholar : PubMed/NCBI
3	Kathju S, Nistico L, Hall-Stoodley L, Post JC, Ehrlich GD and Stoodley P: Chronic surgical site infection due to suture-associated polymicrobial biofilm. Surg Infect (Larchmt). 10:457–461. 2009. View Article : Google Scholar : PubMed/NCBI
4	Sun J, Daniel R, Wagner-Döbler I and Zeng AP: Is autoinducer-2 a universal signal for interspecies communication: A comparative genomic and phylogenetic analysis of the synthesis and signal transduction pathways. BMC Evol Biol. 4:362004. View Article : Google Scholar : PubMed/NCBI
5	Li M, Villaruz AE, Vadyvaloo V, Sturdevant DE and Otto M: AI-2-dependent gene regulation in Staphylococcus epidermidis. BMC Microbiol. 8:42008. View Article : Google Scholar : PubMed/NCBI
6	Taga ME, Semmelhack JL and Bassler BL: The LuxS-dependent autoinducer AI-2 controls the expression of an ABC transporter that functions in AI-2 uptake in Salmonella typhimurium. Mol Microbiol. 42:777–793. 2001. View Article : Google Scholar : PubMed/NCBI
7	Wiesener MS, Turley H, Allen WE, Willam C, Eckardt KU, Talks KL, Wood SM, Gatter KC, Harris AL, Pugh CW, et al: Induction of endothelial PAS domain protein-1 by hypoxia: Characterization and comparison with hypoxia-inducible factor-1alpha. Blood. 92:2260–2268. 1998.PubMed/NCBI
8	Ikeda E, Achen MG, Breier G and Risau W: Hypoxia-induced transcriptional activation and increased mRNA stability of vascular endothelial growth factor in C6 glioma cells. J Biol Chem. 270:19761–19766. 1995. View Article : Google Scholar : PubMed/NCBI
9	Agarwala V, Choudhary N and Gupta S: A risk-benefit assessment approach to selection of adjuvant chemotherapy in elderly patients with early breast cancer: A mini review. Indian J Med Paediatr Oncol. 38:526–534. 2017. View Article : Google Scholar : PubMed/NCBI
10	Shan W and Que HF: The effect of qingre baidu mixture on the biofilm of Staphylococcus aureus and Pseudomonas aeruginosa in chronic and refractory wounds. Guiding J Trad Chin Med Pharm. 23:12–16. 2017.
11	Driver VR and Blume PA: Evaluation of wound care and health-care use costs in patients with diabetic foot ulcers treated with negative pressure wound therapy versus advanced moist wound therapy. J Am Podiatr Med Assoc. 104:147–153. 2014. View Article : Google Scholar : PubMed/NCBI
12	Price LB, Liu CM, Melendez JH, Frankel YM, Engelthaler D, Aziz M, Bowers J, Rattray R, Ravel J, Kingsley C, et al: Community analysis of chronic wound bacteria using 16S rRNA gene-based pyrosequencing: Impact of diabetes and antibiotics on chronic wound microbiota. PLoS One. 4:e64622009. View Article : Google Scholar : PubMed/NCBI
13	Rondas AA, Schols JM, Stobberingh EE and Halfens RJ: Prevalence of chronic wounds and structural quality indicators of chronic wound care in Dutch nursing homes. Int Wound J. 12:630–635. 2015. View Article : Google Scholar : PubMed/NCBI
14	Camilli A and Bassler BL: Bacterial small-molecule signaling pathways. Science. 311:1113–1116. 2006. View Article : Google Scholar : PubMed/NCBI
15	Miller MB and Bassler BL: Quorum sensing in bacteria. Annu Rev Microbiol. 55:165–199. 2001. View Article : Google Scholar : PubMed/NCBI
16	Hentzer M, Wu H, Andersen JB, Riedel K, Rasmussen TB, Bagge N, Kumar N, Schembri MA, Song Z, Kristoffersen P, et al: Attenuation of Pseudomonas aeruginosa virulence by quorum sensing inhibitors. EMBO J. 22:3803–3815. 2003. View Article : Google Scholar : PubMed/NCBI
17	Scutera S, Zucca M and Savoia D: Novel approaches for the design and discovery of quorum-sensing inhibitors. Expert Opin Drug Discov. 9:353–366. 2014. View Article : Google Scholar : PubMed/NCBI
18	Sztajer H, Lemme A, Vilchez R, Schulz S, Geffers R, Yip CY, Levesque CM, Cvitkovitch DG and Wagner-Döbler I: Autoinducer-2-regulated genes in Streptococcus mutans UA159 and global metabolic effect of the luxS mutation. J Bacteriol. 190:401–415. 2008. View Article : Google Scholar : PubMed/NCBI
19	Auger S, Krin E, Aymerich S and Gohar M: Autoinducer 2 affects biofilm formation by Bacillus cereus. Appl Environ Microbiol. 72:937–941. 2006. View Article : Google Scholar : PubMed/NCBI
20	Bachtiar EW, Bachtiar BM, Jarosz LM, Amir LR, Sunarto H, Ganin H, Meijler MM and Krom BP: AI-2 of Aggregatibacter actinomycetemcomitans inhibits Candida albicans biofilm formation. Front Cell Infect Microbiol. 4:942014. View Article : Google Scholar : PubMed/NCBI
21	Azakami H, Teramura I, Matsunaga T, Akimichi H, Noiri Y, Ebisu S and Kato A: Characterization of autoinducer 2 signal in Eikenella corrodens and its role in biofilm formation. J Biosci Bioeng. 102:110–117. 2006. View Article : Google Scholar : PubMed/NCBI
22	González Barrios AF, Zuo R, Hashimoto Y, Yang L, Bentley WE and Wood TK: Autoinducer 2 controls biofilm formation in Escherichia coli through a novel motility quorum-sensing regulator (MqsR, B3022). J Bacteriol. 188:305–316. 2006. View Article : Google Scholar : PubMed/NCBI
23	Huang Z, Meric G, Liu Z, Ma R, Tang Z and Lejeune P: luxS-based quorum-sensing signaling affects Biofilm formation in Streptococcus mutans. J Mol Microbiol Biotechnol. 17:12–19. 2009. View Article : Google Scholar : PubMed/NCBI
24	Geier H, Mostowy S, Cangelosi GA, Behr MA and Ford TE: Autoinducer-2 triggers the oxidative stress response in mycobacterium avium, leading to biofilm formation. Appl Environ Microbiol. 74:1798–1804. 2008. View Article : Google Scholar : PubMed/NCBI
25	Oh SY, Kwon HC, Kim SH, Jang JS, Kim MC, Kim KH, Han JY, Kim CO, Kim SJ, Jeong JS and Kim HJ: Clinicopathologic significance of HIF-1alpha, p53, and VEGF expression and preoperative serum VEGF level in gastric cancer. BMC Cancer. 8:1232008. View Article : Google Scholar : PubMed/NCBI
26	Cane G, Ginouvès A, Marchetti S, Buscà R, Pouysségur J, Berra E, Hofman P and Vouret-Craviari V: HIF-1alpha mediates the induction of IL-8 and VEGF expression on infection with Afa/Dr diffusely adhering E. coli and promotes EMT-like behaviour. Cell Microbiol. 12:640–653. 2010. View Article : Google Scholar : PubMed/NCBI
27	Yang M, Jing L, Wang J, Yu Y, Cao L, Zhang L, Zhou X and Sun Z: Macrophages participate in local and systemic inflammation induced by amorphous silica nanoparticles through intratracheal instillation. Int J Nanomedicine. 11:6217–6228. 2016. View Article : Google Scholar : PubMed/NCBI
28	Cuffini AM, Carlone NA, Xerri L and Pizzoglio MF: Synergy of ceftazidime and human macrophages on phagocytosis and killing of Staphylococcus aureus and Pseudomonas aeruginosa. J Antimicrob Chemother. 20:261–271. 1987. View Article : Google Scholar : PubMed/NCBI
29	Brubaker AL, Rendon JL, Ramirez L, Choudhry MA and Kovacs EJ: Reduced neutrophil chemotaxis and infiltration contributes to delayed resolution of cutaneous wound infection with advanced age. J Immunol. 190:1746–1757. 2013. View Article : Google Scholar : PubMed/NCBI
30	Gander S: Bacterial biofilms: Resistance to antimicrobial agents. J Antimicrob Chemother. 37:1047–1050. 1996. View Article : Google Scholar : PubMed/NCBI
31	Ghigo JM: Natural conjugative plasmids induce bacterial biofilm development. Nature. 412:442–445. 2001. View Article : Google Scholar : PubMed/NCBI
32	Chew SC, Yam JKH, Matysik A, Seng ZJ, Klebensberger J, Givskov M, Doyle P, Rice SA, Yang L and Kjelleberg S: Matrix polysaccharides and SiaD diguanylate cyclase alter community structure and competitiveness of Pseudomonas aeruginosa during dual-species biofilm development with Staphylococcus aureus. MBio. 9(pii): e00585–18. 2018.PubMed/NCBI