噬菌体对小鼠泛耐药鲍曼不动杆菌脓毒症疗效的研究
发布时间：2018-08-30 10:32
【摘要】：研究背景和目的目前鲍曼不动杆菌已成为医院内感染的重要致病菌之一,而且在烧伤病房尤其突出,笔者所在科室烧伤重症病房2011年1月至2014年12月收治的162例烧伤患者的血液标本共计1658份,病原菌检出339株,前三位分别是鲍曼不动杆菌(Acinetobacter baumannii,ABA),金黄色葡萄球菌(Staphylococcus aureus,SAU)以及铜绿假单胞菌(Pseudomonas aeruginosa,PAE)。国内大部分文献报道鲍曼不动杆菌的检出率仅低于铜绿假单胞菌,位居第二。据国外文献报道,1991年纽约首次暴发鲍曼不动杆菌感染以来,世界各地对鲍曼不动杆菌耐药现象的报道层出不穷,尤其是对碳青霉烯类抗菌素耐药现象的增多,使成功治疗鲍曼不动杆菌所致感染变得困难。加上开发新的抗菌素周期长,耗费资金巨大以及细菌耐药变异迅速,亟需寻找新的治疗方案。噬菌体是一种以细菌为宿主的病毒,噬菌体制剂经过临床验证能够有效治疗细菌感染。1958年,我国细菌学教授余贺,就利用噬菌体成功治疗了铜绿假单胞菌所致烧伤患者感染。过去几十年里,大量抗生素得以研发并广泛应用于临床,相应限制了噬菌体的使用。近年来,随着细菌耐药情况越来越严重,欧美国家开始重新重视噬菌体在细菌感染治疗中的作用,国内一些单位也开始进行相关研究。笔者单位在筛选分离噬菌体并建立噬菌体库方面进行探索。本研究通过筛选泛耐药鲍曼不动杆菌裂解性噬菌体,建立泛耐药鲍曼不动杆菌所致小鼠脓毒症模型,并使用所筛选到的噬菌体进行相关的动物实验。通过分析噬菌体治疗后小鼠存活情况、对白细胞计数的影响及其细菌清除情况,观察和研究噬菌体对来源于烧伤患者的泛耐药鲍曼不动杆菌所致小鼠脓毒症的治疗效果。研究方法(1)泛耐药鲍曼不动杆菌裂解性噬菌体的分离与保存使用混合宿主菌扩增噬菌体的方法,以泛耐药鲍曼不动杆菌为宿主菌,在笔者所在医院未经处理的污水池中取污水筛选得到泛耐药鲍曼不动杆菌裂解性噬菌体8株,并测定各株裂解性噬菌体的裂解谱,选取其裂解谱最宽、裂解效果最好的裂解性噬菌体备用,作为抗菌剂进行后续动物治疗实验。(2)小鼠泛耐药鲍曼不动杆菌脓毒症模型的建立将48只8~12周龄雄性健康BALB/c小鼠,随机分为4组,每组12只,记为A,B,C,D四组。分别向A,B,C,D,四组小鼠腹腔注射泛耐药鲍曼不动杆菌5×108 CFU/mL,2.5×108 CFU/mL,1×108 CFU/mL,5×107 CFU/mL这四个浓度的菌液各1mL。注射成功后,将各组小鼠按正常饲养,密切观察各组小鼠生命体征,死亡小鼠数量,腹腔灌洗液细菌培养结果,血培养结果。(3)噬菌体对小鼠泛耐药鲍曼不动杆菌脓毒症的治疗效果Ⅰ.将60只BALB/c小鼠,按随机数字表法分为空白对照组、脓毒症对照组、抗生素治疗组、噬菌体治疗组、噬菌体对照组,每组12只。空白对照组小鼠腹腔(注射部位下同)注射生理盐水1 mL;脓毒症对照组、抗生素治疗组、噬菌体治疗组小鼠注射5×107 CFU/mL鲍曼不动杆菌1 mL建立脓毒症模型。2 h后,脓毒症对照组注射生理盐水1 mL,抗生素治疗组注射1 mg/mL亚胺培南/西司他丁1 mL,噬菌体治疗组腹腔注射1×108 PFU/mL噬菌体1 m L;噬菌体对照组注射1×108 PFU/mL噬菌体1 mL。每组均连续注射7 d,每天观察记录小鼠存活情况。Ⅱ.另取60只BALB/c小鼠,同Ⅰ分组处理。实验第2、4、6天当天注射前,每组选1~3只存活小鼠,每只鼠取眶静脉血20μL检查白细胞计数。实验第2天取血小鼠另取1 mL眶静脉血行细菌培养;另取小鼠肺、肝、肾、脾组织,匀浆、稀释后行菌落计数后计算细菌含量。对数据行Wilcoxon秩和检验、单因素方差分析、LSD检验及Kruskal-Wallis秩和检验。结果(1)以22株泛耐药鲍曼不动杆菌为宿主菌,笔者所在医院未经消毒处理的污水池为噬菌体来源,共筛选得到8株裂解性噬菌体,通过噬菌体裂解谱测定,其中Bp201404072号噬菌体裂解谱59%,高于其他7株噬菌体,故选取该株噬菌体作为抗菌剂,用于后续动物实验,4℃冰箱冻存。(2)各组小鼠注射不同浓度鲍曼不动杆菌后,四组小鼠均出现典型的脓毒症相关症状,其中A,B两组小鼠较C,D两组急性感染症状明显。实验第7天,A,B,C,D四组小鼠均死亡。其中A组小鼠实验第3天均死亡,B组小鼠于实验第4天全部死亡,C组小鼠于实验第5天全部死亡,D组小鼠于实验第7天均死亡(其中一只小鼠存活时予以处死,取眶静脉血做细菌培养)。各组小鼠生存天数采用log-rank test,与D组相比,其他各组存活天数较短(P=0.002,有统计学差异)。A,B,C,D四组小鼠腹腔灌洗液细菌培养可见明显菌落形成。D组存活小鼠眶静脉血培养结果提示阳性,细菌学鉴定结果提示为鲍曼不动杆菌。对比各组实验结果,D组注射菌液浓度(5×107CFU/mL)符合后续动物实验要求。故后续小鼠泛耐药鲍曼不动杆菌脓毒症模型菌液浓度定为5×107 CFU/mL。(3)Ⅰ.实验第7天,空白对照组、脓毒症对照组、抗生素治疗组、噬菌体治疗组、噬菌体对照组小鼠存活数分别为12、0、8、10、12只。与脓毒症对照组比较,其余4组小鼠存活比均明显升高(Z值为55.635~106.593,P值均小于0.05);噬菌体治疗组小鼠存活率稍高于抗生素治疗组,但差异无统计学意义(Z=2.797,P0.05)。Ⅱ.脓毒症对照组、抗生素治疗组、噬菌体治疗组小鼠实验第2天分别死亡3、2、1只,实验第4天分别死亡10、7、6只,实验第6天分别死亡12、11、8只。实验第2天,空白对照组、噬菌体治疗组、噬菌体对照组小鼠白细胞计数相近,3组均明显低于脓毒症对照组的(P0.05),且后2组明显低于抗生素治疗组的(P值均小于0.05)。实验第4天,抗生素治疗组、噬菌体治疗组、噬菌体对照组小鼠白细胞计数相近,明显低于空白对照组(P0.05),前4组白细胞计数均明显低于脓毒症对照组(P0.01)。实验第6天,空白对照组、抗生素治疗组、噬菌体治疗组、噬菌体对照组小鼠白细胞计数差异无统计学意义(χ2=4.128,P0.05)。实验第2天,空白对照组、脓毒症对照组、抗生素治疗组、噬菌体治疗组、噬菌体对照组分别有0、12、7、2、0只小鼠血细菌培养结果为阳性。空白对照组、噬菌体治疗组、噬菌体对照组小鼠血细菌培养阳性比显著低于脓毒症对照组(χ2值为-30.0~30.0,P值均小于0.01)。抗生素治疗组小鼠血细菌培养阳性比显著高于空白对照组和噬菌体对照组(χ2值分别为17.5、-17.5,P值均小于0.05)。实验第2天,除噬菌体治疗组小鼠肾组织外,空白对照组、噬菌体治疗组、噬菌体对照组小鼠各脏器组织细菌含量均为0,均显著低于脓毒症对照组(χ2值为-9.0~9.0,P值均小于0.01);抗生素治疗组小鼠肾组织细菌含量显著高于空白对照组和噬菌体对照组(χ2值分别为-7.5、7.5,P值均小于0.05)。结论通过筛选得到的泛耐药鲍曼不动杆菌裂解性噬菌体作为抗菌剂,用于噬菌体治疗小鼠泛耐药鲍曼不动杆菌所致脓毒症,能明显提高脓毒症小鼠生存率,控制炎症反应,证明了泛耐药鲍曼不动杆菌噬菌体运用于临床治疗相关细菌感染的可行性。
[Abstract]:BACKGROUND AND OBJECTIVE Acinetobacter baumannii has become one of the most important pathogens of nosocomial infections, especially in burn wards. 162 blood samples from 162 burn patients admitted to our department from January 2011 to December 2014 were collected. 339 strains of pathogens were detected. The first three strains of Acinetobacter baumannii were immobile. Acinetobacter baumannii (ABA), Staphylococcus aureus (SAU) and Pseudomonas aeruginosa (PAE). Most domestic literatures reported that the detection rate of Acinetobacter baumannii was only lower than Pseudomonas aeruginosa, ranking second. According to foreign literature reports, the first outbreak of Acinetobacter baumannii in New York in 1991. Since the infection of Acinetobacter baumannii, reports of drug resistance to Acinetobacter baumannii have emerged all over the world. Especially, the increasing resistance to carbapenems makes it difficult to successfully treat Acinetobacter baumannii infection. Phage is a bacterial-hosted virus that has been proved to be effective in the treatment of bacterial infections. In 1958, Yu He, a professor of bacteriology in China, successfully treated burn patients with Pseudomonas aeruginosa infection by using phages. In the past few decades, a large number of antibiotics have been developed and widely used. In recent years, with the bacterial drug resistance becoming more and more serious, European and American countries began to attach importance to the role of phages in the treatment of bacterial infections, and some domestic units began to carry out relevant research. To establish a model of sepsis in mice induced by pan-resistant Acinetobacter baumannii by screening the lysing phages of pan-resistant Acinetobacter baumannii and to carry out related animal experiments with the selected phages. Methods (1) Isolation and preservation of lysis phages of pan-resistant Acinetobacter baumannii using mixed host bacteria to amplify phages. Pan-resistant Acinetobacter baumannii was used as host bacteria and was not found anywhere in our hospital. Eight strains of pan-resistant Acinetobacter baumannii lysing phages were screened out from the sewage in a rational sewage tank, and the lysing spectra of each strain were determined. The lysing phages with the broadest lysing spectra and the best lysing effect were selected as reserve for subsequent animal treatment as antimicrobial agents. (2) Pan-resistant Acinetobacter baumannii sepsis in mice Forty-eight healthy male BALB/c mice aged from 8 to 12 weeks were randomly divided into four groups, 12 mice in each group, which were divided into four groups: A, B, C and D. The mice in each group were injected intraperitoneally with pan-drug-resistant Acinetobacter baumannii 5 *108 CFU/mL, 2.5 *108 CFU/mL, 1 *108 CFU/mL and 5 *107 CFU/mL, respectively. The vital signs, the number of dead mice, the results of bacterial culture in peritoneal lavage fluid and blood culture were observed. (3) The therapeutic effect of bacteriophage on Pan-drug-resistant Acinetobacter baumannii sepsis in mice I. 60 BALB/c mice were divided into blank control group, sepsis control group and antibiotic treatment group according to random number table. The mice in blank control group were injected with 1 mL normal saline, the mice in sepsis control group, the mice in antibiotic treatment group, and the mice in phage treatment group were injected with 5 *107 CFU/mL Acinetobacter baumannii to establish sepsis model. The treatment group was injected with 1 mg/mL imipenem/cilastatin 1 mL, the phage treatment group was injected with 1 x 108 PFU/mL phage 1 mL, and the phage control group was injected with 1 x 108 PFU/mL phage 1 mL. Before irradiation, 1-3 mice in each group were selected, and 20 mu L of orbital vein blood was taken from each mouse to check the white blood cell count.On the second day of the experiment, another 1 mL of orbital vein blood was taken from the blood of the mice for bacterial culture.The lung, liver, kidney, spleen tissue, homogenate of the mice were taken and the bacterial content was counted after dilution.The data were tested by Wilcoxon rank sum test, one-way ANOVA and LSD. Kruskal-Wallis rank sum test was performed. Results (1) A total of 8 lysing phages were obtained from 22 pan-resistant Acinetobacter baumannii strains as host bacteria and non-disinfected sewage pools in our hospital. The lysis spectrum of phage Bp2014072 was 59%, which was higher than that of the other 7 strains. The bacteriophage was used as an antimicrobial in the following animal experiments and frozen at 4 C for storage. (2) After injecting different concentrations of Acinetobacter baumannii into each group of mice, typical sepsis-related symptoms were found in the four groups. The acute infection symptoms of A, B, C and D mice were more obvious than those of C and D mice. All the mice in group B died on the 3rd day, all the mice in group B died on the 4th day, all the mice in group C died on the 5th day, and all the mice in group D died on the 7th day of the experiment (one of the mice was killed while surviving, and the orbital vein blood was taken for bacterial culture). The bacterial culture of peritoneal lavage fluid of mice in groups A, B, C and D showed obvious colony formation. The blood culture of orbital vein of surviving mice in group D was positive, and the result of bacteriological identification was Acinetobacter baumannii. (3) On the 7th day of the experiment, there were 12, 0, 8, 10 and 12 mice in the blank control group, sepsis control group, antibiotic treatment group, bacteriophage treatment group and bacteriophage control group, respectively. The survival rate of mice in the phage treatment group was slightly higher than that in the antibiotic treatment group, but there was no significant difference (Z = 2.797, P 0.05). II. Sepsis control group, antibiotic treatment group and phage treatment group died 3, 2, 1 mice on the second day, 10, 7, 6 mice on the fourth day, respectively. On the second day of the experiment, the white blood cell counts of the blank control group, the phage treatment group and the phage control group were similar. The white blood cell counts of the three groups were significantly lower than those of the sepsis control group (P 0.05), and the latter two groups were significantly lower than those of the antibiotic treatment group (P < 0.05). The white blood cell count of the control group was similar to that of the blank control group (P 0.05). The white blood cell count of the first four groups was significantly lower than that of the sepsis control group (P 0.01). On the sixth day of the experiment, the white blood cell count of the blank control group, the antibiotic treatment group, the phage treatment group and the phage control group had no significant difference (2=4.128, P 0.05). There were 0,12,7,2,0 mice in the blank control group, sepsis control group, antibiotic treatment group, phage treatment group and bacteriophage control group respectively. The positive rate of blood bacterial culture in the blank control group, phage treatment group and bacteriophage control group was significantly lower than that in the sepsis control group (_2 value - 30.0-30.0, P value was less than 0. The positive rate of blood bacterial culture in antibiotic treatment group was significantly higher than that in blank control group and bacteriophage control group (_2 value was 17.5, -17.5, P value was less than 0.05 respectively). On the second day of experiment, the bacterial content in all organs of mice except the kidney tissue of bacteriophage treatment group, blank control group, bacteriophage treatment group and bacteriophage control group were all 0. It was significantly lower than the sepsis control group (_2 value was - 9.0-9.0, P value was less than 0.01); the bacterial content of kidney tissue in the antibiotic treatment group was significantly higher than that in the blank control group and the phage control group (_2 value was - 7.5, 7.5, P value was less than 0.05). Conclusion The pan-drug-resistant Acinetobacter baumannii lysis phages were screened as antibiotics and used as antibacterial agents. Phage therapy for sepsis caused by pan-drug-resistant Acinetobacter baumannii in mice can significantly improve the survival rate of septic mice and control inflammatory reaction, which proves the feasibility of pan-drug-resistant Acinetobacter baumannii phage in clinical treatment of related bacterial infections.
【学位授予单位】：第三军医大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R644
本文编号：2212807
[Abstract]:BACKGROUND AND OBJECTIVE Acinetobacter baumannii has become one of the most important pathogens of nosocomial infections, especially in burn wards. 162 blood samples from 162 burn patients admitted to our department from January 2011 to December 2014 were collected. 339 strains of pathogens were detected. The first three strains of Acinetobacter baumannii were immobile. Acinetobacter baumannii (ABA), Staphylococcus aureus (SAU) and Pseudomonas aeruginosa (PAE). Most domestic literatures reported that the detection rate of Acinetobacter baumannii was only lower than Pseudomonas aeruginosa, ranking second. According to foreign literature reports, the first outbreak of Acinetobacter baumannii in New York in 1991. Since the infection of Acinetobacter baumannii, reports of drug resistance to Acinetobacter baumannii have emerged all over the world. Especially, the increasing resistance to carbapenems makes it difficult to successfully treat Acinetobacter baumannii infection. Phage is a bacterial-hosted virus that has been proved to be effective in the treatment of bacterial infections. In 1958, Yu He, a professor of bacteriology in China, successfully treated burn patients with Pseudomonas aeruginosa infection by using phages. In the past few decades, a large number of antibiotics have been developed and widely used. In recent years, with the bacterial drug resistance becoming more and more serious, European and American countries began to attach importance to the role of phages in the treatment of bacterial infections, and some domestic units began to carry out relevant research. To establish a model of sepsis in mice induced by pan-resistant Acinetobacter baumannii by screening the lysing phages of pan-resistant Acinetobacter baumannii and to carry out related animal experiments with the selected phages. Methods (1) Isolation and preservation of lysis phages of pan-resistant Acinetobacter baumannii using mixed host bacteria to amplify phages. Pan-resistant Acinetobacter baumannii was used as host bacteria and was not found anywhere in our hospital. Eight strains of pan-resistant Acinetobacter baumannii lysing phages were screened out from the sewage in a rational sewage tank, and the lysing spectra of each strain were determined. The lysing phages with the broadest lysing spectra and the best lysing effect were selected as reserve for subsequent animal treatment as antimicrobial agents. (2) Pan-resistant Acinetobacter baumannii sepsis in mice Forty-eight healthy male BALB/c mice aged from 8 to 12 weeks were randomly divided into four groups, 12 mice in each group, which were divided into four groups: A, B, C and D. The mice in each group were injected intraperitoneally with pan-drug-resistant Acinetobacter baumannii 5 *108 CFU/mL, 2.5 *108 CFU/mL, 1 *108 CFU/mL and 5 *107 CFU/mL, respectively. The vital signs, the number of dead mice, the results of bacterial culture in peritoneal lavage fluid and blood culture were observed. (3) The therapeutic effect of bacteriophage on Pan-drug-resistant Acinetobacter baumannii sepsis in mice I. 60 BALB/c mice were divided into blank control group, sepsis control group and antibiotic treatment group according to random number table. The mice in blank control group were injected with 1 mL normal saline, the mice in sepsis control group, the mice in antibiotic treatment group, and the mice in phage treatment group were injected with 5 *107 CFU/mL Acinetobacter baumannii to establish sepsis model. The treatment group was injected with 1 mg/mL imipenem/cilastatin 1 mL, the phage treatment group was injected with 1 x 108 PFU/mL phage 1 mL, and the phage control group was injected with 1 x 108 PFU/mL phage 1 mL. Before irradiation, 1-3 mice in each group were selected, and 20 mu L of orbital vein blood was taken from each mouse to check the white blood cell count.On the second day of the experiment, another 1 mL of orbital vein blood was taken from the blood of the mice for bacterial culture.The lung, liver, kidney, spleen tissue, homogenate of the mice were taken and the bacterial content was counted after dilution.The data were tested by Wilcoxon rank sum test, one-way ANOVA and LSD. Kruskal-Wallis rank sum test was performed. Results (1) A total of 8 lysing phages were obtained from 22 pan-resistant Acinetobacter baumannii strains as host bacteria and non-disinfected sewage pools in our hospital. The lysis spectrum of phage Bp2014072 was 59%, which was higher than that of the other 7 strains. The bacteriophage was used as an antimicrobial in the following animal experiments and frozen at 4 C for storage. (2) After injecting different concentrations of Acinetobacter baumannii into each group of mice, typical sepsis-related symptoms were found in the four groups. The acute infection symptoms of A, B, C and D mice were more obvious than those of C and D mice. All the mice in group B died on the 3rd day, all the mice in group B died on the 4th day, all the mice in group C died on the 5th day, and all the mice in group D died on the 7th day of the experiment (one of the mice was killed while surviving, and the orbital vein blood was taken for bacterial culture). The bacterial culture of peritoneal lavage fluid of mice in groups A, B, C and D showed obvious colony formation. The blood culture of orbital vein of surviving mice in group D was positive, and the result of bacteriological identification was Acinetobacter baumannii. (3) On the 7th day of the experiment, there were 12, 0, 8, 10 and 12 mice in the blank control group, sepsis control group, antibiotic treatment group, bacteriophage treatment group and bacteriophage control group, respectively. The survival rate of mice in the phage treatment group was slightly higher than that in the antibiotic treatment group, but there was no significant difference (Z = 2.797, P 0.05). II. Sepsis control group, antibiotic treatment group and phage treatment group died 3, 2, 1 mice on the second day, 10, 7, 6 mice on the fourth day, respectively. On the second day of the experiment, the white blood cell counts of the blank control group, the phage treatment group and the phage control group were similar. The white blood cell counts of the three groups were significantly lower than those of the sepsis control group (P 0.05), and the latter two groups were significantly lower than those of the antibiotic treatment group (P < 0.05). The white blood cell count of the control group was similar to that of the blank control group (P 0.05). The white blood cell count of the first four groups was significantly lower than that of the sepsis control group (P 0.01). On the sixth day of the experiment, the white blood cell count of the blank control group, the antibiotic treatment group, the phage treatment group and the phage control group had no significant difference (2=4.128, P 0.05). There were 0,12,7,2,0 mice in the blank control group, sepsis control group, antibiotic treatment group, phage treatment group and bacteriophage control group respectively. The positive rate of blood bacterial culture in the blank control group, phage treatment group and bacteriophage control group was significantly lower than that in the sepsis control group (_2 value - 30.0-30.0, P value was less than 0. The positive rate of blood bacterial culture in antibiotic treatment group was significantly higher than that in blank control group and bacteriophage control group (_2 value was 17.5, -17.5, P value was less than 0.05 respectively). On the second day of experiment, the bacterial content in all organs of mice except the kidney tissue of bacteriophage treatment group, blank control group, bacteriophage treatment group and bacteriophage control group were all 0. It was significantly lower than the sepsis control group (_2 value was - 9.0-9.0, P value was less than 0.01); the bacterial content of kidney tissue in the antibiotic treatment group was significantly higher than that in the blank control group and the phage control group (_2 value was - 7.5, 7.5, P value was less than 0.05). Conclusion The pan-drug-resistant Acinetobacter baumannii lysis phages were screened as antibiotics and used as antibacterial agents. Phage therapy for sepsis caused by pan-drug-resistant Acinetobacter baumannii in mice can significantly improve the survival rate of septic mice and control inflammatory reaction, which proves the feasibility of pan-drug-resistant Acinetobacter baumannii phage in clinical treatment of related bacterial infections.
【学位授予单位】：第三军医大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R644
【参考文献】
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