| 摘要: |
| 目的研究白介素35(IL-35)蛋白在小鼠铜绿假单胞肺炎中的保护作用。方法雄性健康清洁级昆明小鼠90只,随机分成正常对照组(生理盐水组:30只)、肺部感染组(PA组:30只)、干预组(30只)。PA组:造模前12h自腹腔注射生理盐水1mL,戊巴比妥钠(50mg/kg)麻醉小鼠后固定,待麻药显效后,常规消毒铺巾,依次切开小鼠颈部皮肤、皮下、皮下组织,显露气管上段,从气管环状软骨间隙内穿刺注入铜绿假单胞菌(PA)菌液0、03mL(2个麦氏单位浓度即6×108cfu/mL),以建立小鼠肺炎模型;干预组:在造模前12h自腹腔注射白介素35蛋白1mL(含白介素35蛋白2μg),肺炎模型建立同PA组;正常对照组:操作同前两组,在造模前12h自腹腔注射生理盐水1mL,最后从气管环状软骨间隙内穿刺注入生理盐水0.03mL作为正常对照。造模成功后,三组均在第3、9、24h随机抓取10只小鼠,留置标本后处死小鼠。取各个实验组在不同时间点小鼠右肺肺泡灌洗液,经离心后,用酶联免疫吸附法(enzyme-linked immunosorbnent assay,ELISA)检测各组肺泡灌洗液中IL-17、IL-8的浓度;取左肺大部分肺组织经甲醛固定后,行HE染色比较肺炎炎症的范围和程度,取左肺小部分肺组织,用于制作肺组织匀浆并行细菌培养及菌落计数。结果感染组、干预组小鼠肺组织在各个时间点存在明显的炎症改变,以24h组炎症改变最明显,但干预组在3、9、24h炎症改变明显轻于感染组,两者比较差异有统计学意义(P<0.05);干预组各时间组IL-17,IL-8表达水平明显低于感染组,且差异有统计学意义(P<0.05)。结论 (1)IL-35对小鼠铜绿假单胞菌肺炎具有保护作用,可明显减轻其炎症反应。(2)IL-35减轻小鼠铜绿假单胞菌肺炎炎症反应可能与抑制炎性介质IL-8,IL-17有关。 |
| 关键词: 白介素35 铜绿假单胞菌 炎症 |
| DOI： |
| 分类号: |
| 基金项目: |
|
| Protective Effects of IL-35 on Pneumonia in Mice caused by Pseudomonas Aeruginosa |
|
Tang Yujian;Zhan Xiaoqin;1,2
|
|
1.Respiratory Department of the People’s Hospital of RongXian County;2.The Affiliated Hospital of Southwest Medical University
|
| Abstract: |
| Objective To study the protectiue effect of IL-35 protein on pseudomonas aeruginosa pneumonia in mice.Methods 90 healthy and clean male kunming mice were randomly divided into normal control group( normal saline,n = 30),pulmonary infection group( group PA,n = 30) and the intervention group( n = 30). Group PA: Firstly the mice received injection of physiological saline 1 ml through intraperitoneal 12 hours before modeling. Secondly the mice were anesthetized with pentobarbital sodium( 50 mg/kg) and then was fixed. After the anesthesia had taken effect,we conventionally disinfected,shopped towels and successively cut the skin,subcutaneous,subcutaneous tissue and exposed upper trachea of mice. And then we injected pseudomonas aeruginosa fluid 0. 03 mL( 2 Maxwell unit concentration,or 6 × 10~8 cfu/mL) by puncture of cricoid cartilage gap of trachea puncture,in order to establish the mouse pneumonia model; The intervention group: Firstly the mice received injection of interleukin( including IL-35 protein 2 ug) through intraperitoneal 12 hours before modeling,and the next steps were the same as the PA group. Normal control group: Operations were the same as 2 groups above. The mice received injection of physiological saline 1 ml through intraperitoneal 12 hours before modeling,and last we injected physiological saline 0. 03 ml by puncture of cricoid cartilage gap of trachea puncture as normal control. After succeeding modeling,10 mice were randomly selected respectively after 3,9 and24 hours,and were killed after taking samples. The bronchoalveolar lavage fluids which came from the right lung of micewere collected at different time points in each group. And after they were centrifugal separated,the concentrations of IL-8 and IL-17 in bronchoalveolar lavage fluid in each group were tested by enzyme-linked immunosorbent assay( ELISA). The most of the left lung tissue was taken and fixed by 10% paraformaldehyde,and stained by HE staining to compare the range and degree of inflammation.A small part of left lung tissue was taken to make lung tissue homogenate which was used for bacterial culture and colony counting.Results The lung tissues of mice in the infection group and the intervention group had obvious inflammatory changes at each time point,particularly inflammatory change at 24 h was the most obvious while inflammatory changes of the intervention group at 3,9,24 hours were significantly lighter than the infection group,and the comparison between the two groups had significant difference( P<0. 05). 2. The expression levels of IL-8 and IL-17 of the intervention group at different time points were significantly lower than that of the infection group,and there was a statistically significant difference( P<0. 05). Conclusions IL-35 has a protective effect on mice with pseudomonas aeruginosa pneumonia and can obviously reduce the inflammatory process. IL-35 attenuating pseudomonas aeruginosa pneumonia inflammation may be related to the inhibition of inflammatory mediators IL-8 and IL-17. |
| Key words: IL-35 pseudomonasaeruginosa inflammation |
