Reversal of antibiotic resistance methicillanin-resistant staphylococcus aureus by polyethyleneimine-phosphothioate oligodeoxynucleotide nanometer particle

Abstract

Abstract: AIM: This study used polyethyleneimine (PEI) and phosphothioate oligodeoxynucleotide (PS-ODNs) to prepare PEI-ODNs nanometer particle and transmit it into methicillin resistant staphylococcus aureus (MRSA) in vitro by pharmacodynamic evaluation. METHODS: The PEI-ODNs nanometer particle was prepared and its diameter was analyzed. The PEI and PS-ODNs binding rate by fluorospectrophotometer was evaluated. The total colony forming unit (CFU) was counted. The cell growth curve was drawn by measuring A630 nm at different time points with microplate reader. The minimal inhibitory concentration (MIC) was determined by fluid dilution method. RESULTS: The diameter of PEI-ODNs nanometer particle was (85±22) nm and the highest binding rate was (97.3±1.1) %. In M-H agar plate(including 6 μg/mL oxacillin), the number of MRSA colonies was 6.4×108 mL in 30 μg/mL PEI-ODNs nanometer particle treated group, while the number of MRSA colonies in blank control was 3.3×109 mL. The number of MRSA colonies was significantly decreased in PEI-ODNs nanometer particle treated group (P<0.01), while CFU of MRSA was not influenced in control group (P >0.05). Significant growth inhibition of cells treated with PEI-ODNs nanometer particle was observed as compared with those cells in control. The MIC of oxacillin to MRSA was decreased from 1 024 μg/mL to 16 μg/mL when treated with 30 μg/mL PEI-ODNs nanometer particle. CONCLUSION: PEI binds PS-ODNs with high binding rate and has little diameter. The susceptibility of MRSA to β-lactam antibiotics is significantly restored. The results indicate that the PEI can transmit PS-ODNs into MRSA and it is considered as a new vector for PS-ODNs into bacteria.

Key words: phosphothioate oligodeoxynucleotide, polyethyleneimine, nanometer particle, methicillin resistant staphylococcus aureus, transfection

CLC Number:

R966

WANG Hui, MENG Jing-ru, CHEN Tao, JIA Ming, HE Gong-hao, MA Xue, LUO Xiaoxing. Reversal of antibiotic resistance methicillanin-resistant staphylococcus aureus by polyethyleneimine-phosphothioate oligodeoxynucleotide nanometer particle[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2008, 13(1): 31-35.

References

[1] Hiramatsu K, Hanaki H, Ino T, et al. Methicillin- resistant staphylococcus aureus clinical strain with reduced vancomycin susceptibity[J]. J Antimicrob Chemother, 1997,40(2): 135-136.
[2] Archer GL, Bosilevac JM. Signaling Antibiotic Resistance in Staphylococci[J]. Science, 2001, 291(9): 1915-1916.
[3] Crooke ST. Oligonucleotide analogs might be designed to bind to mRNA[J]. Anticancer Drug Des, 1997, 12(5): 311-313.
[4] 孟静茹, 罗晓星, 扈本荃, 等. 特异性阻断MecR1-MecI-MecA 通路逆转耐甲氧西林金黄色葡萄球菌耐药性[J]. 第四军医大学学报, 2005, 26(15): 1352-1355.
[5] Meng JR, Hu BQ, Liu J, et al. Restoration of oxacillin susceptibility in methicillin-resistant staphylococcus aureus by blocking the MecR1-mediated signaling pathway[J]. J Chemother, 2006, 18(4): 88-93.
[6] Zhou Z, Meng JR, Niu C, et al. Restoration of antibiotic susceptibility in methicillinresistant staphylococcus aureus by targeting mecr1 with a phosphorothioate deoxyribozyme[J]. Clin Exp Pharmacol Physiol, 2007, 34(11): 1160-1164.
[7] Lungwitz U, Breuning M, Blunk T, et al. Polyethyleneimine-based non-viral gene delivery system[J]. Eur J Pharm Biopharm, 2005, 60(2): 247-266.
[8] ThomasM, Klibanov AM. Non-viral gene therapy: polycation-mediated DNA delivery[J]. Appl Microbiol Biotechnol,2003, 62(1): 27-34.
[9] Boussif O, Lezoualc'h F, Zanta MA, et al. A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethyleneimine[J]. Proc Natl Acad Sci USA, 1995, 92(16): 7297-7301.
[10] Torres Viera C, Tsiodras S, Gold HS, et al. Restoration of Vancomycin susceptibility in Enterococcus faecalis by antiresistance determinant gene transfer[J]. Antimicrob Agents Chemother, 2001, 45(3): 973-975.
[11] Harth G, Zamecnik PC, Tang JY, et al. Treatment of Mycobaterium tuberculosis with antisense oligonucleotides to glutamine synthetase mRNA inhibits glutamine synthetase activity, formation of the poly-L-glutamate glutamine cell wall struture, and bacterial replication[J]. Proc Natl Acad Sci USA, 2000, 97(1): 418-423.