黄连提取物含药血清对MRSA毒力因子表达的影响
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摘要
目的研究不同浓度黄连含药血清对耐甲氧西林金黄色葡萄球菌毒力因子的表达情况,阐明黄连对MRSA活性的抑制作用机制,为临床应用黄连治疗MRSA感染提供依据。方法选取5株MRSA临床分离株,大鼠灌胃给予黄连提取物后制备含药血清,采用与MRSA体外共培养16h,分为空白血清组、6mg·kg~(-1)组、12mg·kg~(-1)组、24mg·kg~(-1)组的黄连含药血清培养。采用荧光定量PCR方法检测各组MRSA株毒力相关基因葡萄球菌蛋白A(spa)、α-溶血素(hlα)mRNA相对表达水平。结果无抗菌活性的黄连含药血清与MRSA体外共培养16h,2个MRSA毒力因子spa和hla的mRNA相对表达水平平均低于对照组(P<0.01)。结论黄连可作为一种潜在的抗MRSA感染药物,可进一步开发。
OBJECTIVE To explore the effect of different concentrations of serum containing coptis on the genetic expression of methicillin-resistant Staphylococcus aureus(MRSA) virulence factor and illuminate their mechanisms,and to provide the basis for using coptis.METHODS Five MRSA clinical isolates were selected and were cultured with drug-containing serum of mouse which was prepared after intragastric administration with coptis for 16 h.which were divided into blank serum(control group) and serum contained 6 mg·kg~(-1),12 mg·kg~(-1),24 mg·kg~(-1) of coptis(serum containing coptis with low,medium and high concentration groups).Fluorescent quantitative real-time PCR method was used to determine the relative mRNA expression levels of virulence-related genes staphylococcal protein A(spa),α-hemolysin(Hla) in various groups.RESULTS After co-culture for 16 h,different concentrations of serum containing coptis without antibacterial activity were significantly decreased compared with control group(P<0.01).CONCLUSION The extracts from coptis are potentially useful in developing anti-MRSA drugs.
OBJECTIVE To explore the effect of different concentrations of serum containing coptis on the genetic expression of methicillin-resistant Staphylococcus aureus(MRSA) virulence factor and illuminate their mechanisms,and to provide the basis for using coptis.METHODS Five MRSA clinical isolates were selected and were cultured with drug-containing serum of mouse which was prepared after intragastric administration with coptis for 16 h.which were divided into blank serum(control group) and serum contained 6 mg·kg~(-1),12 mg·kg~(-1),24 mg·kg~(-1) of coptis(serum containing coptis with low,medium and high concentration groups).Fluorescent quantitative real-time PCR method was used to determine the relative mRNA expression levels of virulence-related genes staphylococcal protein A(spa),α-hemolysin(Hla) in various groups.RESULTS After co-culture for 16 h,different concentrations of serum containing coptis without antibacterial activity were significantly decreased compared with control group(P<0.01).CONCLUSION The extracts from coptis are potentially useful in developing anti-MRSA drugs.
引文
[1]吴静,王克霞,胡联华.黄连对幽门螺杆菌的体外抗菌活性研究[J].时珍国医国药,2006,17(12),2486-2487.
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[7]Welte T,Pletz MW.Antimicrobial treatment of nosocomial meticillin-resistant Staphylococcus aureus(MRSA) pneumonia:current and future options[J].Int J Antimicrob Agents,2010,36:391-410.
[8]Karampela I,Poulakou G,Dimopoulos G.Community acquired methicilin resistant Staphylococcus aureus pneuomonia:an update for the emergency and intensive care physician[J].Minerva Anestesiol,2012,78(8):930-940.
[9]Francois VG,Thomas H.Staphylococcus aureus hemolysins,bi-component liukocidins,and cytolyticpeptides:are dundantarsenal of membrane-damaging virulence factors[J].Cellular and Infection Microbiology,2012,2:120-146.
[10]Dinges PM,Schlievert PM.Exotoxins of Staphylococcus aureus[J].Clin Mierobio Rev,2000,3(1):16-34.
[11]Jayasinghe L,Miles G,Bayley H.Role of the anino latch of staphylococcalalpha-hemolysin in pore formation:a cooperative interaction between the N-terminus and position 217[J].Biol Chem,2006,281(4):2195-2204.
[12]Schmitt CK,Meysick KC,Brien AD.Bacterial Toxins:Friends or Fose[J].Emerg Infect Dis,1999,5(2):224-334.
[13]Wardenburg JB,Patel RJ,Schneewind O.Surface proteins and exotoxins are required for the pathogenesis of Staphylococcus aureus pneumonia[J].2007,75:1040-1044.
[14]Wardenburg JB,Schneewind O.Vaccine protection against Staphylococcus aureus pneumonia[J].J Exp Med,2008,205:287-294.
[15]Kobayashi SD,DeLio FR.Staphylococcus aureus protein A promotes immune suppression[J].mBio,2013,4(5):e00764-13.
[2]林健,林志立,张小玲.黄连提取物含药血清抑制MRSA α-溶血素分泌活性研究[J].九江学院学报,2017,32(2):96-98.
[3]Nygaard TK1,Pallister KB,Ruzevich P,et al.SaeR binds a consensus sequence within virulence gene promoters to advance USA300 pathogenesis[J].J Infect Dis,2010,201(2):241-254.
[4]Iwama H,Amagaya S,Ogihara Y,Effect of shosaikoto,a Japanese and Chinese traditional herbal medicinal mixture,on the mitogenic activity of lipopolysaccharide:a new pharmacological testing method[J].J Ethnopharmacol,1987,21(1):45-53.
[5]陈智能,苏友新,杨连梓.疾病模型血清药理学及其在骨代谢研究中的若干问题[J].中国骨伤,2007,20(1):41-43.
[6]刘建勋,韩笑,孙宇扬.含药血清药理作用强度与体内给药量效时效关系研究[J].中国中药杂志,2006,31(10):829-831.
[7]Welte T,Pletz MW.Antimicrobial treatment of nosocomial meticillin-resistant Staphylococcus aureus(MRSA) pneumonia:current and future options[J].Int J Antimicrob Agents,2010,36:391-410.
[8]Karampela I,Poulakou G,Dimopoulos G.Community acquired methicilin resistant Staphylococcus aureus pneuomonia:an update for the emergency and intensive care physician[J].Minerva Anestesiol,2012,78(8):930-940.
[9]Francois VG,Thomas H.Staphylococcus aureus hemolysins,bi-component liukocidins,and cytolyticpeptides:are dundantarsenal of membrane-damaging virulence factors[J].Cellular and Infection Microbiology,2012,2:120-146.
[10]Dinges PM,Schlievert PM.Exotoxins of Staphylococcus aureus[J].Clin Mierobio Rev,2000,3(1):16-34.
[11]Jayasinghe L,Miles G,Bayley H.Role of the anino latch of staphylococcalalpha-hemolysin in pore formation:a cooperative interaction between the N-terminus and position 217[J].Biol Chem,2006,281(4):2195-2204.
[12]Schmitt CK,Meysick KC,Brien AD.Bacterial Toxins:Friends or Fose[J].Emerg Infect Dis,1999,5(2):224-334.
[13]Wardenburg JB,Patel RJ,Schneewind O.Surface proteins and exotoxins are required for the pathogenesis of Staphylococcus aureus pneumonia[J].2007,75:1040-1044.
[14]Wardenburg JB,Schneewind O.Vaccine protection against Staphylococcus aureus pneumonia[J].J Exp Med,2008,205:287-294.
[15]Kobayashi SD,DeLio FR.Staphylococcus aureus protein A promotes immune suppression[J].mBio,2013,4(5):e00764-13.