Krupple样因子6调控诱导型一氧化氮合酶介导铜绿假单胞菌感染小鼠肺组织细胞凋亡的研究  

KLF6 and iNOS Regulated Apoptosis During Pseudomonas Aeruginosa Lung Infection in Mice

作　　者：喻志芳 韩彬 柴文戍 

机构地区：[1]锦州医科大学附属第一医院呼吸内科,辽宁省锦州市121001

出　　处：《中国全科医学》2016年第20期2435-2439,共5页Chinese General Practice

基　　金：辽宁省自然科学基金资助项目(2014022015;20092189)

摘　　要：目的通过研究Krupple样因子6（KLF6）在铜绿假单胞菌感染肺组织中表达水平的变化,探讨KLF6调控诱导型一氧化氮合酶（iNOS）介导铜绿假单胞菌感染小鼠肺组织细胞凋亡的机制。方法于2014年10—12月,30只小鼠随机分为对照组和低感染组,每组15只,低感染组注射铜绿假单胞菌1.0×10~7CFU制作动物模型,感染后3、9、24 h,分别随机取各组5只小鼠留取标本。另10只小鼠随机分为中感染组和高感染组,每组5只,分别注射铜绿假单胞菌5.0×10~7、1.0×10~8CFU,均在感染后9 h留取标本。在各时间点处死动物后无菌分离肺组织,左肺结扎后测定湿干重之比。采用TUNEL法检测肺组织细胞凋亡指数,采用硝酸还原酶法测定一氧化氮（NO）水平,化学比色法测定iNOS活性,采用Western blotting法检测肺组织KLF6水平。结果光镜下可见低感染组小鼠肺组织大量炎性细胞浸润,肺泡隔增厚,肺泡壁结构破坏,局部有肺泡膨胀不全或肺泡塌陷,并扩散到肺间质内,24 h时炎症改变最为严重。3 h时,对照组和低感染组肺组织湿干重之比比较,差异无统计学意义（P〉0.05）;9、24 h时,低感染组肺组织湿干重之比大于对照组,差异有统计学意义（P〈0.05）。低感染组不同时间肺组织凋亡指数比较,差异有统计学意义（P〈0.05）。不同剂量铜绿假单胞菌感染9 h后肺组织凋亡指数比较,差异有统计学意义（P〈0.05）。3、9、24 h时,低感染组肺组织NO水平、iNOS活性均高于对照组,差异有统计学意义（P〈0.05）。不同剂量铜绿假单胞菌感染9 h后肺组织NO水平、iNOS活性比较,差异均有统计学意义（P〈0.05）。低感染组不同时间肺组织KLF6表达水平比较,差异有统计学意义（P〈0.05）。不同剂量铜绿假单胞菌感染9 h后肺组织KLF6表达水平比较,差异均有统计学意义（P〈0.05）。结论铜绿假单胞菌感染能诱导KLF6的表达,并有可能通过iNOS调控介�Objective To research the changes of KLF6 expression in lung tissue with pseudomonas aeruginosa infection and investigate the mechanism of KLF6 and iNOS regulated apoptosis during pseudomonas aeruginosa lung infection in mice. Methods From October to December in 2014,30 mice were randomly divided into control group and low- infection group,with 15 mice in each group. The low- infection group was injected with pseudomonas aeruginosa by 1. 0 × 10^7 CFU to make models; 5 mice were randomly selected respectively at 3,9 and 24 hours. Other 10 mice were divided into medium-infection group and high- infection group,with 5 mice in each group. Medium- infection group and high- infection group were infected with pseudomonas aeruginosa by 5. 0 × 10^7 CFU and 1. 0 × 10^8 CFU respectively,and samples were obtained 9 hours later. At each time point mentioned above,animals were killed to perform sterile separation of their lung tissue,and litigation was conducted on the left lung to determine wet and dry weight ratio. Lung tissue apoptosis index was detected using TUNEL method,and NO level was determined using nitrale reduetase method,iNOS activity was determined by chemical colorimetry. The KLF6 expression of lung tissue was detected by Western blotting. Results Using the microscope,we observed much inflammatory cell infiltration,thickened alveolar septa,destroyed structure of the alveolar wall,and local alveolar atelectasis or alveolar collapse spreading to pulmonary interstitium; the inflammatory changes became the severest at 24 h. At 3 h,control group and low-infection group were not significantly different in wet and dry weight ratio（ P 〉0. 05）; at 9 and 24 h,low- infection group was higher than control group in wet and dry weight ratio（ P 〈0. 05）. There were significant differences in the lung tissue apoptosis index of low- infection group among different time points（ P〈 0. 05）. There were significant differences in the lung tissue apoptosis index among different dosages of pseudomonas aeruginosa inf

关 键 词：铜绿假单胞菌 Krupple样因子6 一氧化氮合酶 细胞凋亡 

分 类 号：R378.991[医药卫生—病原生物学]