17-AAG联合卡泊芬净治疗中性粒细胞缺乏大鼠侵袭性肺曲霉菌病的效果
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摘要
目的探讨Hsp90抑制剂17-烯丙基-17脱甲氧基格尔德霉素(17-AAG)联合卡泊芬净(CAS)治疗中性粒细胞缺乏的侵袭性曲霉菌病(IPA)的效果。方法将40只Wistar雄性大鼠按随机数字表法分为4组,即A组(空白对照)、B组、C组及D组,每组10只。四组大鼠均用环磷酰胺腹腔注射制备中性粒细胞缺乏模型,再经鼻吸入烟曲霉孢子悬液。A组正常饲养,作为空白对照,B组和C组分别给予1 mg/kg的CAS和1 mg/kg的17-AAG腹腔注射,D组给予17-AAG联合CAS(用法、用量同上),共用药10 d。记录大鼠的生存状况。检测治疗第3天及第7天大鼠的血清半乳糖甘露醇聚糖抗原(GM),并对肺组织进行GMS染色,观察感染部位的真菌负荷。结果与A组比较,B组及D组生存率均明显较高(P <0.01),且D组平均生存时间长于B组,但差异无统计学意义(P>0.05)。与A组比较,治疗第3天各组GM值差异无统计学意义(P> 0.05);治疗第7天,各治疗组GM值低于A组(P <0.01),D组低于B组(P <0.05)。肺组织切片GMS染色显示,各治疗组真菌感染病灶均减小,且D组减小最为显著,其治疗第15天未发现病灶。结论 17-AAG可通过抑制应激反应来提高CAS对大鼠IPA的疗效。
Objective To explore the effects of the combination of Hsp90 inhibitor 17-AAG(17-allyl-17-demethoxygeldanamycin) and Caspofungin(CAS) in the treatmemt of invasive aspergillosis(IPA) with neutrophil deficiency. Methods Forty male Wistar rats were divided into 4 groups, group A, group B, group C and group D, according to random number table method, with 10 rats in each group. Neutrophil deficiency models were made by intraperitoneal injection of Cyclophosphamide in four groups of rats, and then Aspergillus fumigatus spore suspension was inhaled through nose.Group A was fed normally and served as blank control. Group B and C were given CAS of 1 mg/kg and 17-AAG of 1 mg/kg respectively by intraperitoneal injection. Group D was given 17-AAG combined with CAS(the same usage and dosage)for 10 days. The survival status of rats was recorded. The serum galactomannan antigen(GM) was detected on the 3 rd and 7 th day after treatment, and the lung tissues were stained with GMS to observe the fungal load at the infected sites.Results Compared with group A, the survival rates of group B and group D were significantly increased(P < 0.01), and the average survival time of group D was longer than that of group B, but the difference was not statistically significant(P > 0.05). Compared with group A, there was no significant difference in GM values between the groups on the third day of treatment(P > 0.05). On the 7 th day of treatment, the GM values of each treatment group were lower than those of group A(P < 0.01), while group D was lower than group B(P < 0.05). GMS staining of lung slices showed that the lesions of fungal infection decreased in all treatment groups, and the most significant reduction was found in group D.No lesions were found on the 15 th day of treatment. Conclusion 17-AAG improves the efficacy of CAS in the treatment of IPA by inhibiting stress response.
Objective To explore the effects of the combination of Hsp90 inhibitor 17-AAG(17-allyl-17-demethoxygeldanamycin) and Caspofungin(CAS) in the treatmemt of invasive aspergillosis(IPA) with neutrophil deficiency. Methods Forty male Wistar rats were divided into 4 groups, group A, group B, group C and group D, according to random number table method, with 10 rats in each group. Neutrophil deficiency models were made by intraperitoneal injection of Cyclophosphamide in four groups of rats, and then Aspergillus fumigatus spore suspension was inhaled through nose.Group A was fed normally and served as blank control. Group B and C were given CAS of 1 mg/kg and 17-AAG of 1 mg/kg respectively by intraperitoneal injection. Group D was given 17-AAG combined with CAS(the same usage and dosage)for 10 days. The survival status of rats was recorded. The serum galactomannan antigen(GM) was detected on the 3 rd and 7 th day after treatment, and the lung tissues were stained with GMS to observe the fungal load at the infected sites.Results Compared with group A, the survival rates of group B and group D were significantly increased(P < 0.01), and the average survival time of group D was longer than that of group B, but the difference was not statistically significant(P > 0.05). Compared with group A, there was no significant difference in GM values between the groups on the third day of treatment(P > 0.05). On the 7 th day of treatment, the GM values of each treatment group were lower than those of group A(P < 0.01), while group D was lower than group B(P < 0.05). GMS staining of lung slices showed that the lesions of fungal infection decreased in all treatment groups, and the most significant reduction was found in group D.No lesions were found on the 15 th day of treatment. Conclusion 17-AAG improves the efficacy of CAS in the treatment of IPA by inhibiting stress response.
引文
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[20]Li J,Soroka J,Buchner J.The Hsp90 chaperone machinery;conformational dynamics and regulation by cochaperones[J].Acta Biochim Biophys Sin,2012,1823(3):624-635.
[2]Snelders E,Melchers WJ,Verweij PE.Azole resistance in Aspergillus fumigatus:a new challenge in the management of invasive aspergillosis[J].Future Microbiol,2011,6(3):335-347.
[3]De Pauw B,Walsh TJ,Donnelly JP,et al.Revised definitions of invasive fungal disease from the European Organization for Research and Treatment of Cancer/Invasive Fungal Infections Cooperative Group and the National Institute of Allergy and Infectious Diseases Mycoses Study Group(EORTC/MSG)Consensus Group[J].Clin Infect Dis,2008,46(12):1813-1821.
[4]Patterson TF,Thompson GR,Denning DW,et al.Practice Guidelines for the Diagnosis and Management of Aspergillosis:2016 Update by the Infectious Diseases Society of America[J].Clin Infect Dis,2016,63(4):1-60.
[5]Morrison VA.Echinocandin antifungals:review and update[J]Expert Rev Anti Infect Ther,2006,4(2):325-342.
[6]Maschmeyer G,Glasmacher A.Pharmacological properties and clinical efficacy of a recently licensed systemic antifungal,caspofungin[J].Mycoses,2005,48(4):227-234
[7]Cowen LE,Steinbach WJ.Stress,drugs,and evolution:the role of cellular signaling in fungal drug resistance[J].Eukaryotic Cell,2008,7(5):747-764.
[8]Cowen LE.The evolution of fungal drug resistance:modulating the trajectory from genotype to phenotype[J].Nat Rev Microbiol,2008,6(3):187-198.
[9]Leach MD,Klipp E,Cowen LE,et al.Fungal Hsp90:abiological transistor that tune scelular outputs to thermal inputs[J].Nat Rev Microbiol,2012,10(10):693-704.
[10]Mehrad B,Strieter RM,Standiford TJ.Role of TNF-αin pulmonary host defense in murine invasive Aspergillosis[J]J Immunol,1999,162(3):1633-1640.
[11]王妍,龙飞,祁卉卉,等.烟曲霉菌孢子对哮喘大鼠气道炎症和气道反应性的影响[J].中国呼吸与危重监护杂志,2007,6(3):185-190.
[12]Havaux X,Zeine A,Dits A,et al.A new mouse model of lung allergy induced by the spores of Alternaria alternate and Cladosporium herbarum molds[J].Clin Exp Immunol,2004,139(2):179-188.
[13]Denis O,van den Brule S,Heymans J,et al.Chronic intranasal administration of mould spores or extracts to unsensitized mice leads to lung allergic inflammation,hyperreactivity and remodeling[J].Immunology,2007,122(2):268-278.
[14]Kontoyiannis DP,Marr KA,Park BJ,et al.Prospective surveillance for invasive fungal infections in hematopoietic stem cell transplant recipients,2001-2006:overview of the Transplant-Associated Infection Surveillance Network(TRANSNET)Database[J].Clin Infect Dis,2010,50(8):1091-1100.
[15]Pappas PG,Alexander BD,Andes DR,et al.Invasive fungal infections among organ transplant recipients:results of the Transplant Associate Infection Surveillance Network(TRANSNET)[J].Clin Infect Dis,2010,50(8):1101-1111.
[16]Trepel J,Mollapour M,Giaccone G,et al.Targeting the dynamic HSP90 complex in cancer[J].Nat Rev Cancer,2010,10(8):537-549.
[17]Whitesell L,Lindquist SL.HSP90 and the chaperoning of cancer[J].Nat Rev Cancer,2005,5(10):761-772.
[18]Fortwendel JR,Juvvadi PR,Pinchai N,et al.Differential effects of inhibiting chitin and 1,3-{beta}-D-glucan synthesis in ras and calcineurin mutants of Aspergillus fumigatus[J].Antimicrob Agents Chemother,2009,53(2):476-482.
[19]Li J,Buchner J.Structure,function and regulation of the hsp90 machinery[J].Biomed J,2013,36(3):106-117.
[20]Li J,Soroka J,Buchner J.The Hsp90 chaperone machinery;conformational dynamics and regulation by cochaperones[J].Acta Biochim Biophys Sin,2012,1823(3):624-635.