中国感染与化疗杂志
中國感染與化療雜誌
중국감염여화료잡지
CHINESE JOURNAL OF INFECTION AND CHEMOTHERAPY
2015年
1期
51-56
,共6页
鲍曼不动杆菌%气管插管%肺炎%动物模型
鮑曼不動桿菌%氣管插管%肺炎%動物模型
포만불동간균%기관삽관%폐염%동물모형
Acinetobacter baumannii%intubation%pneumonia%animal model
目的：建立耳窥镜直视下气管插管法，并采用上述方法构建小鼠鲍曼不动杆菌肺炎模型。方法①采用美国Braintree 公司插管工具盒建立在耳窥镜直视下小鼠气管插管方法；②24只 ICR 雌性小鼠随机分为正常对照组、免疫缺陷组、免疫缺陷感染组，每组8只，其中免疫缺陷组和免疫缺陷感染组采用环磷酰胺腹腔注射后造成免疫缺陷。3组小鼠在试验当日均按上述方法行气管插管后，免疫缺陷感染组沿气管内插管注射10μL 鲍曼不动杆菌（3．2×108 CFU／mL），正常对照组和免疫缺陷组沿气管内插管注射10μL 生理盐水；每组小鼠均分别于0 h 和48 h 眼眶后静脉丛取血，观察白细胞、中性粒细胞、淋巴细胞、单核细胞等计数。并在上述时间点各处死4只小鼠，进行小鼠肺组织菌落计数和肺组织病理检查。结果①10只小鼠行耳窥镜直视下气管插管，均获成功，无小鼠插管后死亡。沿气管内插管注射菌液浓度为3．0×108 CFU／mL，气管插管后0 h 处死小鼠取肺并定量，其菌落计数为2．91×107～5．32×107 CFU／g 肺，平均值±标准差为（4．05×107±0．82×107） CFU／g 肺，表明该方法重复性好；②3组小鼠观察48 h，正常对照组和免疫缺陷组小鼠无死亡，免疫缺陷感染组有2只小鼠死亡（2／8）。与正常对照组比较免疫缺陷组和免疫缺陷感染组小鼠在0 h 均出现血液白细胞和中性粒细胞计数平均值明显下降（P ＜0．01）；免疫缺陷感染组在感染后即刻（0 h）肺内菌量平均值达4．13×107 CFU／g 肺，48 h 小鼠肺内菌量平均值显著升高达到3．62×1010 CFU／g 肺，与0 h 比较菌量增长近1000倍（P ＜0．01），免疫缺陷感染组病理检查结果显示肺组织内局限性肉芽肿形成，肺泡腔内可见脓肿形成。结论采用耳窥镜直视下气管插管法构建小鼠鲍曼不动杆菌肺炎模型操作简便，成功率高，且注射入小鼠肺内的菌量较恒定，试验重复性好，本研究成功构建了小鼠鲍曼不动杆菌肺炎模型。
目的：建立耳窺鏡直視下氣管插管法，併採用上述方法構建小鼠鮑曼不動桿菌肺炎模型。方法①採用美國Braintree 公司插管工具盒建立在耳窺鏡直視下小鼠氣管插管方法；②24隻 ICR 雌性小鼠隨機分為正常對照組、免疫缺陷組、免疫缺陷感染組，每組8隻，其中免疫缺陷組和免疫缺陷感染組採用環燐酰胺腹腔註射後造成免疫缺陷。3組小鼠在試驗噹日均按上述方法行氣管插管後，免疫缺陷感染組沿氣管內插管註射10μL 鮑曼不動桿菌（3．2×108 CFU／mL），正常對照組和免疫缺陷組沿氣管內插管註射10μL 生理鹽水；每組小鼠均分彆于0 h 和48 h 眼眶後靜脈叢取血，觀察白細胞、中性粒細胞、淋巴細胞、單覈細胞等計數。併在上述時間點各處死4隻小鼠，進行小鼠肺組織菌落計數和肺組織病理檢查。結果①10隻小鼠行耳窺鏡直視下氣管插管，均穫成功，無小鼠插管後死亡。沿氣管內插管註射菌液濃度為3．0×108 CFU／mL，氣管插管後0 h 處死小鼠取肺併定量，其菌落計數為2．91×107～5．32×107 CFU／g 肺，平均值±標準差為（4．05×107±0．82×107） CFU／g 肺，錶明該方法重複性好；②3組小鼠觀察48 h，正常對照組和免疫缺陷組小鼠無死亡，免疫缺陷感染組有2隻小鼠死亡（2／8）。與正常對照組比較免疫缺陷組和免疫缺陷感染組小鼠在0 h 均齣現血液白細胞和中性粒細胞計數平均值明顯下降（P ＜0．01）；免疫缺陷感染組在感染後即刻（0 h）肺內菌量平均值達4．13×107 CFU／g 肺，48 h 小鼠肺內菌量平均值顯著升高達到3．62×1010 CFU／g 肺，與0 h 比較菌量增長近1000倍（P ＜0．01），免疫缺陷感染組病理檢查結果顯示肺組織內跼限性肉芽腫形成，肺泡腔內可見膿腫形成。結論採用耳窺鏡直視下氣管插管法構建小鼠鮑曼不動桿菌肺炎模型操作簡便，成功率高，且註射入小鼠肺內的菌量較恆定，試驗重複性好，本研究成功構建瞭小鼠鮑曼不動桿菌肺炎模型。
목적：건립이규경직시하기관삽관법，병채용상술방법구건소서포만불동간균폐염모형。방법①채용미국Braintree 공사삽관공구합건립재이규경직시하소서기관삽관방법；②24지 ICR 자성소서수궤분위정상대조조、면역결함조、면역결함감염조，매조8지，기중면역결함조화면역결함감염조채용배린선알복강주사후조성면역결함。3조소서재시험당일균안상술방법행기관삽관후，면역결함감염조연기관내삽관주사10μL 포만불동간균（3．2×108 CFU／mL），정상대조조화면역결함조연기관내삽관주사10μL 생리염수；매조소서균분별우0 h 화48 h 안광후정맥총취혈，관찰백세포、중성립세포、림파세포、단핵세포등계수。병재상술시간점각처사4지소서，진행소서폐조직균락계수화폐조직병리검사。결과①10지소서행이규경직시하기관삽관，균획성공，무소서삽관후사망。연기관내삽관주사균액농도위3．0×108 CFU／mL，기관삽관후0 h 처사소서취폐병정량，기균락계수위2．91×107～5．32×107 CFU／g 폐，평균치±표준차위（4．05×107±0．82×107） CFU／g 폐，표명해방법중복성호；②3조소서관찰48 h，정상대조조화면역결함조소서무사망，면역결함감염조유2지소서사망（2／8）。여정상대조조비교면역결함조화면역결함감염조소서재0 h 균출현혈액백세포화중성립세포계수평균치명현하강（P ＜0．01）；면역결함감염조재감염후즉각（0 h）폐내균량평균치체4．13×107 CFU／g 폐，48 h 소서폐내균량평균치현저승고체도3．62×1010 CFU／g 폐，여0 h 비교균량증장근1000배（P ＜0．01），면역결함감염조병리검사결과현시폐조직내국한성육아종형성，폐포강내가견농종형성。결론채용이규경직시하기관삽관법구건소서포만불동간균폐염모형조작간편，성공솔고，차주사입소서폐내적균량교항정，시험중복성호，본연구성공구건료소서포만불동간균폐염모형。
Objective To construct a new intubation method with an otoscope and establish a murine model of Acinetobacter baumannii pneumonia with this method.Methods Part I:The Hallowell Intubation Pack for mice (Braintree Scientific Inc., USA)was used to construct a new intubation method with an otoscope.Part II:Twenty-four female ICR mice were randomized into 3 groups including control (group 1),immunosuppression (group 2)and infection after immunosuppression groups (group 3),with 8 mice in each group.The mice were treated with cyclophosphamide (CTX)by peritoneal injection to posterior orbital venous plexus.The total number of white blood cells,the number of neutrophils and the percentage of neutrophils were determined.Four mice were sacrificed at 0 h and 48 h after inoculation in each group.Then the lungs from each mouse were aseptically collected for quantitative culture and histopathology.Results Part I:Ten mice were successfully intubated using the new method and none of the mice was dead.Pulmonary bacterial culture at baseline (0 h)was (2.91×107-5.32×107 )CFU/g tissue,while the mean± standard deviation was (4.05 × 107 ± 0.82 × 107 )CFU/g tissue.The results showed that this new method had a perfect repeatability.Part II:Over 48 h,2 mice were dead in group 3,while no mouse was dead in other 2 groups.For group 3,the average pulmonary bacterial culture was 4.13×107 CFU/g tissue at 0 h and reached 3.62×1010 CFU/g tissue at 48 h (increased appropriate 1 000 times,P <0.01).The histopathologic changes in lung showed local granulomas and abscess in the alveolar space.Conclusions Intubation under the guidance of otoscope had the advantages of high repeatability and easy to operate.Additionally,the method provided stable and consistent bacterial inocula into lungs.The murine model of Acinetobacter baumannii pneumonia was successfully established with a new intubation method under the guidance of otoscope.
