补脾清肺方对耐药铜绿假单胞菌毒力因子的影响
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摘要
目的探讨补脾清肺方治疗下呼吸道耐药铜绿假单胞菌(PA)感染的作用机制,为治疗PA所致下呼吸道感染提供依据。方法 SPF级雄性SD大鼠,体重240～260 g,2～3月龄,共30只,设空白对照组(n=10),不造模,给予等体积生理盐水灌服;模型组(n=10),造模后,给予等体积生理盐水灌服;中药组(n=10),造模后,给予2倍剂量(10.68 g)中药灌服,以PA液注入大鼠气管内的方法建立大鼠下呼吸道感染模型,以补脾清肺方进行干预。测定弹性蛋白酶及绿脓菌素含量。结果在弹性蛋白酶含量上,中药组与模型组比较,中药组弹性蛋白酶含量降低,差异有统计学意义(P <0.05);中药组与空白对照组比较,中药组弹性蛋白酶含量升高,差异无统计学意义(P> 0.05);模型组与空白对照组比较,模型组弹性蛋白酶含量升高,差异有统计学意义(P <0.05)。在绿脓菌素含量上,中药组与模型组比较,中药组绿脓菌素含量升高,差异无统计学意义(P> 0.05);中药组与空白对照组比较,中药组绿脓菌素含量升高,差异无统计学意义(P> 0.05);模型组与空白对照组比较,模型组绿脓菌素含量升高,差异无统计学意义(P> 0.05)。结论补脾清肺方可以显著减轻气道炎性反应,降低弹性蛋白酶的含量,减轻PA侵袭。
Objective To investigate the mechanism of Bupi Qingfei Decoction in the treatment of drug-resistant Pseudomonas aeruginosa(PA) infection of the lower respiratory tract, to provide evidence for the treatment of PA-induced lower respiratory tract infection. Methods A total of 30 SPF male SD rats, weighing 240-260 g, aged 2-3 months.Blank control group(n = 10) was set, no model was made, and equal volume of normal saline was given. In the model group(n = 10), normal saline of equal volume was administered after modeling. The rats in the traditional Chinese medicine group(n = 10) were given 2 times the dose(10.68 g) of traditional Chinese medicine after modeling, and the rats′ lower respiratory tract infection model were established by injecting PA solution into rats′ trachea, given Bupi Qingfei Decocion for intervention. The contents of elastase and pyocyanine were determined. Results In terms of elastase content, compared with the model group, the elastase content in the traditional Chinese medicine group decreased,and the difference was statistically significant(P < 0.05). Compared with the blank control group, elastase content in the traditional Chinese medicine group increased, and the difference was not statistically significant(P > 0.05). Compared with the blank control group, the elastase content in the model group increased, and the difference was statisti-cally significant(P < 0.05). In terms of the content of pyocyanine, compared with the model group, the content of pyocyanine increased in the traditional Chinese medicine group, and the difference was not statistically significant(P > 0.05). Compared with the blank control group, pyocyanine content increased in the TCM group,and the difference was not statistically significant(P > 0.05). Compared with the blank control group, the content of pyocyanine in the model group increased, and the difference was not statistically significant(P > 0.05). Conclusion Bupi Qingfei Decoction can significantly improve airway inflammation, reduce the content of elastase and relieve the PA′s invasion.
Objective To investigate the mechanism of Bupi Qingfei Decoction in the treatment of drug-resistant Pseudomonas aeruginosa(PA) infection of the lower respiratory tract, to provide evidence for the treatment of PA-induced lower respiratory tract infection. Methods A total of 30 SPF male SD rats, weighing 240-260 g, aged 2-3 months.Blank control group(n = 10) was set, no model was made, and equal volume of normal saline was given. In the model group(n = 10), normal saline of equal volume was administered after modeling. The rats in the traditional Chinese medicine group(n = 10) were given 2 times the dose(10.68 g) of traditional Chinese medicine after modeling, and the rats′ lower respiratory tract infection model were established by injecting PA solution into rats′ trachea, given Bupi Qingfei Decocion for intervention. The contents of elastase and pyocyanine were determined. Results In terms of elastase content, compared with the model group, the elastase content in the traditional Chinese medicine group decreased,and the difference was statistically significant(P < 0.05). Compared with the blank control group, elastase content in the traditional Chinese medicine group increased, and the difference was not statistically significant(P > 0.05). Compared with the blank control group, the elastase content in the model group increased, and the difference was statisti-cally significant(P < 0.05). In terms of the content of pyocyanine, compared with the model group, the content of pyocyanine increased in the traditional Chinese medicine group, and the difference was not statistically significant(P > 0.05). Compared with the blank control group, pyocyanine content increased in the TCM group,and the difference was not statistically significant(P > 0.05). Compared with the blank control group, the content of pyocyanine in the model group increased, and the difference was not statistically significant(P > 0.05). Conclusion Bupi Qingfei Decoction can significantly improve airway inflammation, reduce the content of elastase and relieve the PA′s invasion.
引文
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[15]McCarty SM,Cochrane CA,Clegg PD,et al.The role of endogenous and exogenous enzymes in chronic wounds:afocus on the implications of aberrant levels of both host and bacterial proteases in wound healing[J].Wound Repair Regen,2012,20(2):125-136.
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[18]潘婷婷,李镛,刘嘉琳,等.铜绿假单胞菌感应系统基因表达与毒力因子的相关性分析[J].内科理论与实践,2015,10(4):294-298.
[19]Welsh MA,Eibergen NR,Moore JD,et al.Small molecule disruption of quorum sensing cross-regulation in Pseudomonas aeruginosa causes major and unexpected alterations to virulence phenotypes[J].J Am Chem Soc,2015,137(4):1510-1519.
[20]Longo F,Rampioni G,BondìR,et al.A new transcriptional repressor of the Pseudomonas aeruginosa quorum sensing receptor gene lasR[J].PLoS One,2013,8(7):E69554.
[2]Jensen PO,Bjarnsholt T,Phipps R,et al.Rapid necrotic killing of polymor-phonuclear leukocytes is caused by quorum-sensing-controlled production of rhamnolipid by Pseudomonas aeruginosa[J].Microbiology,2007,153(5):1329-1338.
[3]Thaden JT,Lory S,Gardner TS.Quorum-sensing regulation of a copper toxicity system in Pseudomonas aeruginosa[J].J Bacteriol,2010,192(10):2557-2568.
[4]Alayande AB,Aung MM,Kim IS.Correlation Between Quorum Sensing Signal Molecules and Pseudomonas aeruginosa′s Biofilm Development and Virulency[J].Curr Microbiol,2018.doi:10.1007.
[5]陈代杰.微生物药物学[M].2版.北京:化学工业出版社,2008:269-290.
[6]Gupta P,Chhibber S,Harjai K.Subinhibitory concentration of ciprofloxacin targets quorum sensing system of Pseudomonas aeruginosa causing inhibition of biofilm formation&reduction of virulence[J].Indian J Med Res,2016,143(5):643-651.
[7]万毅刚,曹世宏,毕建军,等.大鼠支气管扩张症模型研制[J].上海实验动物科学,2000,20(2):65-68.
[8]Cegelski L,Marshall GR,Eldridge GR,et al.The biology and future prospects of antivirulence therapies[J].Nat Rev Microbiol,2008,6(1):17-27.
[9]Olivier MV,Martin K,Sanda R,et al.Identification of catechin as one of the flavonoids from combretum albiflorum bark extract that reduces the production of QuorumSensing-controlled virulence factors in Pseudomonas aeruginosa PAO1[J].Appl Environ Microb,2010,76(1):243-253.
[10]Francesco I,Francesco M,Cejoice RP,et al.New life for an old drug:the anthelmintic drug niclosamide inhibits Pseudomonas aeruginosa quorumsensing[J].Antimicrob Agents Chemother,2013,57(2):996-1005.
[11]周至品,杨兴秀,叶晓雪.传统药物对铜绿假单胞菌生物被膜作用研究进展[J].中药药理与临床,2015,31(6):230-235.
[12]宋雪萍.补脾清肺汤治疗支气管扩张症(脾虚肺热证)的临床观察[D].北京:北京中医药大学,2015.
[13]樊长征.补中柴前连梅煎减少支气管扩张患者急性加重次数的临床疗效研究[D].北京:中国中医科学院,2016.
[14]樊长征,张文江,任培忠,等.补中柴前连梅煎对支气管扩张症大鼠模型肺气管组织病理学的影响[J].世界中医药,2016,11(11):2370-2374.
[15]McCarty SM,Cochrane CA,Clegg PD,et al.The role of endogenous and exogenous enzymes in chronic wounds:afocus on the implications of aberrant levels of both host and bacterial proteases in wound healing[J].Wound Repair Regen,2012,20(2):125-136.
[16]Beaufort N,Corvazier E,Mlanaoindrou S,et al.Disruption of the endothelial barrier by proteases from the bacterial pathogen Pseudomonas aeruginosa:implication of matrilysis and receptor cleavage[J].PLoS One,2013,8(9):E75708.
[17]Jayaseelan S,Ramaswamy D,Dharmaraj S.Pyocyanin:production,applications,challenges and new insights[J].World J Microbiol Biotechnol,2014,30(4):1159-1168.
[18]潘婷婷,李镛,刘嘉琳,等.铜绿假单胞菌感应系统基因表达与毒力因子的相关性分析[J].内科理论与实践,2015,10(4):294-298.
[19]Welsh MA,Eibergen NR,Moore JD,et al.Small molecule disruption of quorum sensing cross-regulation in Pseudomonas aeruginosa causes major and unexpected alterations to virulence phenotypes[J].J Am Chem Soc,2015,137(4):1510-1519.
[20]Longo F,Rampioni G,BondìR,et al.A new transcriptional repressor of the Pseudomonas aeruginosa quorum sensing receptor gene lasR[J].PLoS One,2013,8(7):E69554.