不同来源白念珠菌感染小鼠前后胞外水解酶活力差异
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摘要
目的了解白念珠菌进入小鼠宿主前后其分泌胞外水解酶能力是否改变。方法挑选分泌型天冬氨酸蛋白酶、磷脂酶、脂肪酶活力无差异的艾滋病、外阴阴道念珠病病患来源和健康人来源白念珠菌,尾静脉注射感染小鼠,分离发病死亡小鼠肾脏菌株,比较感染小鼠前后菌株胞外水解酶活力差异。结果不同来源菌株感染的小鼠28d内死亡速率差异显著(χ2=82.5,P<0.05),从高到低依次为艾滋病、外阴阴道念珠病、健康人来源组;各来源菌株感染小鼠后肾脏分离株分泌型天冬氨酸蛋白酶表达水平皆高于感染小鼠前(F=7.89,P<0.05),不同来源区别显著(F=4.96,P<0.05),从高到低依次为艾滋病、外阴阴道念珠病、健康人来源;感染小鼠前均未曾测得磷脂酶活力的各来源菌株中,在感染小鼠后仅艾滋病、外阴阴道念珠病来源组小鼠肾脏分离株可测得磷脂酶活力,但艾滋病、外阴阴道念珠病来源两组间感染小鼠后肾脏分离株磷脂酶活力无显著差异(F=2.54,P>0.05);各来源白念珠菌感染小鼠前后均未测得脂肪酶活力。结论在宿主体内白念珠菌表达天冬氨酸蛋白酶、磷脂酶能力相较于体外有增强;在不同宿主环境状态下,白念珠菌分泌型天冬氨酸蛋白酶和磷脂酶的潜在表达能力有差异,即宿主免疫状态越弱时,白念珠菌分泌型天冬氨酸蛋白酶、磷脂酶潜在表达能力越强。
Extracellular hydrolytic enzymes have been proved as part of the virulence factors of Candida albicans in vitro.However,how those enzymes work is still unknown when the C.albicans cells' growth environment have been changed from the vitro to the host.Therefore,we analyzed the secreted aspartyl proteinase(SAP),lipase(Lip)and phospholipase(PL)from the C.albicans cells isolated from AIDS,vulvovaginal candidiasis(VVC)patients and normal people.During our study,female KM mice were treated with above C.albicans cells from various sources by tail vein injection,and the C.albicans cells were collected from the mice kidneys when they died.And the activities of extracellular hydrolases produced by the selected strains of C.albicans were compared between in vitro and in vivo of mice respectively.The death rate showed significant difference in the mice treated with C.albicans cells from various sources within 28 d,and the death rate caused by C.albicans isolates from AIDS patients was higher than those form VVC and normal persons.Especially the expression levels of SAP of the strains isolated from mice were higher than in vitro,obvious difference was observed(F=7.89,P<0.05),and the differences were more significant in C.albicans isolates from AIDS patients than those from VVC and normal persons.On the other hand,the PL activity was tested as negative when the C.albicans cells cultured on the artificial media,but this activity changed into positive when C.albicans cells from AIDS and VVC sources grew inside the mice's bodies,which showed no significant difference between these two groups(F=2.54,P>0.05).In addition,the activities of Lip were not detected in all the strains.In conclusion,the secretion amount of SAP and PL of C.albicans in vivo in mice is enhanced than that in vitro.Under different host environmental conditions,the potential expression abilities of SAP and PL of C.albicans are different,that means the weaker the host immune status,the stronger the potential expression ability of SAP and PL of C.albicans.
Extracellular hydrolytic enzymes have been proved as part of the virulence factors of Candida albicans in vitro.However,how those enzymes work is still unknown when the C.albicans cells' growth environment have been changed from the vitro to the host.Therefore,we analyzed the secreted aspartyl proteinase(SAP),lipase(Lip)and phospholipase(PL)from the C.albicans cells isolated from AIDS,vulvovaginal candidiasis(VVC)patients and normal people.During our study,female KM mice were treated with above C.albicans cells from various sources by tail vein injection,and the C.albicans cells were collected from the mice kidneys when they died.And the activities of extracellular hydrolases produced by the selected strains of C.albicans were compared between in vitro and in vivo of mice respectively.The death rate showed significant difference in the mice treated with C.albicans cells from various sources within 28 d,and the death rate caused by C.albicans isolates from AIDS patients was higher than those form VVC and normal persons.Especially the expression levels of SAP of the strains isolated from mice were higher than in vitro,obvious difference was observed(F=7.89,P<0.05),and the differences were more significant in C.albicans isolates from AIDS patients than those from VVC and normal persons.On the other hand,the PL activity was tested as negative when the C.albicans cells cultured on the artificial media,but this activity changed into positive when C.albicans cells from AIDS and VVC sources grew inside the mice's bodies,which showed no significant difference between these two groups(F=2.54,P>0.05).In addition,the activities of Lip were not detected in all the strains.In conclusion,the secretion amount of SAP and PL of C.albicans in vivo in mice is enhanced than that in vitro.Under different host environmental conditions,the potential expression abilities of SAP and PL of C.albicans are different,that means the weaker the host immune status,the stronger the potential expression ability of SAP and PL of C.albicans.
引文
[1]Sardi JCO,Scorzoni L,Bernardi T,et al.Candidaspecies:current epidemiology,pathogenicity,biofilm formation,natural antifungal products and new therapeutic options[J].J Med Microbiol,2013,62(1):10-24.DOI:10.1099/jmm.0.045054-0
[2]马虎,崔凡,林昭春.白念珠菌致病相关的水解酶[J].中国真菌学杂志,2012,7(4):247-51.
[3]Silva S,Negri M,Henriques M,et al.Candida glabrata,Candida parapsilosis and Candida tropicalis:biology,epidemiology,pathogenicity and antifungal resistance[J].FEMS Microbiol Rev,2012,36(2):288-305.DOI:10.1111/j.1574-6976.2011.00278.x
[4]Bouza E,Burillo A,Mu1oz P,et al.Mixed bloodstream infections involving bacteria and Candida spp[J].J Antimicrob Chemoth,2013,68(8):1881-1888.DOI:10.1093/jac/dkt099
[5]Tragiannidis A,Tsoulas C,Kerl K,et al.Invasive candidiasis:update on current pharmacotherapy options and future perspectives[J].Expert Opin Pharmacother,2013,14(11):1515-1528.DOI:10.1517/14656566.2013.805204
[6]Hube B.From commensal to pathogen:stage-and tissuespecific gene expression of Candida albicans[J].Curr Opin Microbiol,2004,7(4):336-341.DOI:10.1016/j.mib.2004.06.003
[7]Tsai PW,Chen YT,Hsu PC,et al.Study of Candida albicans and its interactions with the host:A mini review[J].Biomed,2013,3(1):51-64.DOI:10.1016/j.riam.2013.09.018
[8]H9fs S,Mogavero S,Hube B.Interaction of Candida albicans with host cells:virulence factors,host defense,escape strategies,and the microbiota[J].J Microbiol,2016,54(3):149.DOI:10.1007/s12275-016-5514-0
[9]孙康德,张家胜,陈旭,等.口腔白念珠菌3种致病相关分泌型水解酶检测方法的建立及应用[J].检验医学,2016,31(5):355-358.
[10]Teresa DJ.Role of phospholipases in fungal fitness,pathogenicity,and drug development-lessons from Cryptococcus Neoformans[J].Front Microbiol,2010,1(1):125.DOI:10.3389/fmicb.2010.00125
[11]Gropp K,Schild L,Schindler S,et al.The yeast Candida albicans,evades human complement attack by secretion of aspartic proteases[J].Mol Immunol,2009,47(2/3):465-475.DOI:10.1016/j.molimm.2009.08.019
[12]Ells R,Kilian W,Hugo A,et al.Virulence of south african Candida albicans strains isolated from different clinical samples.[J].Medical Mycol,2014,52(3):246-253.DOI:10.1093/mmy/myt013
[13]Stehr F,Felk A,Kretschmar M,et al.Extracellular hydrolytic enzymes and their relevance during Candida albicans infections.[J].Mycoses,2000,43Suppl 2(2):17.
[14]Richardson JP,Moyes DL.Adaptive immune responses to Candida albicans infection[J].Virulence,2015,6(4):327-337.DOI:10.1080/21505594.2015.1004977
[15]Maccallum DM,Odds FC.Temporal events in the intravenous challenge model for experimental Candida albicans infections in female mice[J].Mycoses,2005,48(3):151-161.DOI:10.1111/j.1439-0507.2005.01121.x
[16]Homma M,Kanbe T,Chibaba H,et al.Detection of intracellular forms of secretory aspartic proteinase in Candida albicans[J].J Gen Microbiol,1992,138(3):627-633.DOI:10.1099/00221287-138-3-627
[17]Price MF,Wilkinson ID,Gentry LO.Plate method for detection of phospholipase activity in Candida albicans[J].Sabouraudia,1982,20(1):7.DOI:10.1099/00221287-138-3-627
[18]Kouker G,Jaeger KE.Specific and sensitive plate assay for bacterial lipases[J].Appl Environ Microb,1987,53(1):211-213.
[19]王梅竹,曹煜,赵亮,等.贵州地区临床酵母菌种类组成和分布[J].山东大学学报(医学版),2014,52(8):94-97.
[20]Staib P,Kretschmar M,Nichterlein T,et al.Differential activation of a Candida albicans virulence gene family during infection[J].PNAS,2000,97(11):6102-6107.DOI:10.1073/pnas.110031497
[21]Kretschmar M,Felk A,Staib P,et al.Individual acid aspartic proteinases(Saps)1-6of Candida albicans are not essential for invasion and colonization of the gastrointestinal tract in mice[J].Microb Pathogenesis,2002,32(2):61-70.DOI:10.1006/mpat.2001.0478
[22]毕爽,胡丹丹,姜远英,等.白念珠菌的高适应性与代谢[J].药学实践杂志,2016,34(2):116-118.
[23]Staniszewska M,Bondaryk M,Piat J,et al.Virulence factors of Candida albicans[J].Przeglad Epidemiologiczny,2012,66(4):629-633.
[24]Gropp K,Schild L,Schindler S,et al.The yeast Candida albicans,evades human complement attack by secretion of aspartic proteases[J].Mol Immunol,2009,47(2/3):465-475.DOI:10.1016/j.molimm.2009.08.019
[25]Naglik JR,Challacombe SJ,Hube B.Candida albicans secreted aspartyl proteinases in virulence and pathogenesis[J].Microbiol Mol Biol Rev,2003,67(3):400-428.DOI:10.1128/MMBR.67.3.400-428.2003
[26]Teresa D J.Role of phospholipases in fungal fitness,pathogenicity,and drug development-lessons from cryptococcus neoformans[J].Front Microbiol,2010,1(1):125.DOI:10.3389/fmicb.2010.00125
[27]Schaller M,Borelli C,Korting HC,et al.Hydrolytic enzymes as virulence factors of Candida albicans[J].Mycoses,2005,48(6):365-377.DOI:10.1111/j.1439-0507.2005.01165.x
[28]Stehr F,Felk A,Gácser A,et al.Expression analysis of the Candida albicans lipase gene family during experimental infections and in patient samples[J].FEMS Yeast Res,2004,4(4/5):401-408.
[29]王芸,宋光耀,刘颐轩,等.脂质肾毒性研究进展[J].医学综述,2014,20(19):3475-3477.
[2]马虎,崔凡,林昭春.白念珠菌致病相关的水解酶[J].中国真菌学杂志,2012,7(4):247-51.
[3]Silva S,Negri M,Henriques M,et al.Candida glabrata,Candida parapsilosis and Candida tropicalis:biology,epidemiology,pathogenicity and antifungal resistance[J].FEMS Microbiol Rev,2012,36(2):288-305.DOI:10.1111/j.1574-6976.2011.00278.x
[4]Bouza E,Burillo A,Mu1oz P,et al.Mixed bloodstream infections involving bacteria and Candida spp[J].J Antimicrob Chemoth,2013,68(8):1881-1888.DOI:10.1093/jac/dkt099
[5]Tragiannidis A,Tsoulas C,Kerl K,et al.Invasive candidiasis:update on current pharmacotherapy options and future perspectives[J].Expert Opin Pharmacother,2013,14(11):1515-1528.DOI:10.1517/14656566.2013.805204
[6]Hube B.From commensal to pathogen:stage-and tissuespecific gene expression of Candida albicans[J].Curr Opin Microbiol,2004,7(4):336-341.DOI:10.1016/j.mib.2004.06.003
[7]Tsai PW,Chen YT,Hsu PC,et al.Study of Candida albicans and its interactions with the host:A mini review[J].Biomed,2013,3(1):51-64.DOI:10.1016/j.riam.2013.09.018
[8]H9fs S,Mogavero S,Hube B.Interaction of Candida albicans with host cells:virulence factors,host defense,escape strategies,and the microbiota[J].J Microbiol,2016,54(3):149.DOI:10.1007/s12275-016-5514-0
[9]孙康德,张家胜,陈旭,等.口腔白念珠菌3种致病相关分泌型水解酶检测方法的建立及应用[J].检验医学,2016,31(5):355-358.
[10]Teresa DJ.Role of phospholipases in fungal fitness,pathogenicity,and drug development-lessons from Cryptococcus Neoformans[J].Front Microbiol,2010,1(1):125.DOI:10.3389/fmicb.2010.00125
[11]Gropp K,Schild L,Schindler S,et al.The yeast Candida albicans,evades human complement attack by secretion of aspartic proteases[J].Mol Immunol,2009,47(2/3):465-475.DOI:10.1016/j.molimm.2009.08.019
[12]Ells R,Kilian W,Hugo A,et al.Virulence of south african Candida albicans strains isolated from different clinical samples.[J].Medical Mycol,2014,52(3):246-253.DOI:10.1093/mmy/myt013
[13]Stehr F,Felk A,Kretschmar M,et al.Extracellular hydrolytic enzymes and their relevance during Candida albicans infections.[J].Mycoses,2000,43Suppl 2(2):17.
[14]Richardson JP,Moyes DL.Adaptive immune responses to Candida albicans infection[J].Virulence,2015,6(4):327-337.DOI:10.1080/21505594.2015.1004977
[15]Maccallum DM,Odds FC.Temporal events in the intravenous challenge model for experimental Candida albicans infections in female mice[J].Mycoses,2005,48(3):151-161.DOI:10.1111/j.1439-0507.2005.01121.x
[16]Homma M,Kanbe T,Chibaba H,et al.Detection of intracellular forms of secretory aspartic proteinase in Candida albicans[J].J Gen Microbiol,1992,138(3):627-633.DOI:10.1099/00221287-138-3-627
[17]Price MF,Wilkinson ID,Gentry LO.Plate method for detection of phospholipase activity in Candida albicans[J].Sabouraudia,1982,20(1):7.DOI:10.1099/00221287-138-3-627
[18]Kouker G,Jaeger KE.Specific and sensitive plate assay for bacterial lipases[J].Appl Environ Microb,1987,53(1):211-213.
[19]王梅竹,曹煜,赵亮,等.贵州地区临床酵母菌种类组成和分布[J].山东大学学报(医学版),2014,52(8):94-97.
[20]Staib P,Kretschmar M,Nichterlein T,et al.Differential activation of a Candida albicans virulence gene family during infection[J].PNAS,2000,97(11):6102-6107.DOI:10.1073/pnas.110031497
[21]Kretschmar M,Felk A,Staib P,et al.Individual acid aspartic proteinases(Saps)1-6of Candida albicans are not essential for invasion and colonization of the gastrointestinal tract in mice[J].Microb Pathogenesis,2002,32(2):61-70.DOI:10.1006/mpat.2001.0478
[22]毕爽,胡丹丹,姜远英,等.白念珠菌的高适应性与代谢[J].药学实践杂志,2016,34(2):116-118.
[23]Staniszewska M,Bondaryk M,Piat J,et al.Virulence factors of Candida albicans[J].Przeglad Epidemiologiczny,2012,66(4):629-633.
[24]Gropp K,Schild L,Schindler S,et al.The yeast Candida albicans,evades human complement attack by secretion of aspartic proteases[J].Mol Immunol,2009,47(2/3):465-475.DOI:10.1016/j.molimm.2009.08.019
[25]Naglik JR,Challacombe SJ,Hube B.Candida albicans secreted aspartyl proteinases in virulence and pathogenesis[J].Microbiol Mol Biol Rev,2003,67(3):400-428.DOI:10.1128/MMBR.67.3.400-428.2003
[26]Teresa D J.Role of phospholipases in fungal fitness,pathogenicity,and drug development-lessons from cryptococcus neoformans[J].Front Microbiol,2010,1(1):125.DOI:10.3389/fmicb.2010.00125
[27]Schaller M,Borelli C,Korting HC,et al.Hydrolytic enzymes as virulence factors of Candida albicans[J].Mycoses,2005,48(6):365-377.DOI:10.1111/j.1439-0507.2005.01165.x
[28]Stehr F,Felk A,Gácser A,et al.Expression analysis of the Candida albicans lipase gene family during experimental infections and in patient samples[J].FEMS Yeast Res,2004,4(4/5):401-408.
[29]王芸,宋光耀,刘颐轩,等.脂质肾毒性研究进展[J].医学综述,2014,20(19):3475-3477.