摘要
目的:探索能使药物基本均匀、稳定地分布于肺部的暴露式气管注射给药方法,并科学设置其对照组。方法:以美蓝代替药物,暴露成年健康小鼠(体质量18~30 g)气管,从干预措施、给药体位、速度、所需的客观药物体积及小鼠主观可耐受体积考虑,予暴露式气管注射。继而以小鼠(体质量20~24 g)为实验对象,以水溶性药物脂多糖(LPS)为例,按上述方法予LPS(2 mg/m L)及0.9%氯化钠溶液建立LPS组及NS组,于3、8、14 h时处死,正常组小鼠无干预,予同时处死,ELISA法检测支气管肺泡灌洗液(BALF)中单核细胞趋化蛋白1(MCP-1)水平,BALF中沉淀的细胞予瑞氏-姬姆萨染色计算中性粒细胞百分比(PMN%),肺组织行病理HE染色及细胞间黏附分子1(ICAM-1)免疫组织化学检测,测定肺组织湿重/干重比(W/D比)。结果:美蓝基本均匀、稳定地分布于小鼠各肺叶;与正常组比,LPS组BALF中的MCP-1水平在3、8 h时明显增加(P<0.05),随时间的推移BALF中PMN%逐渐增加,肺组织炎症改变明显,ICAM-1表达明显增加,W/D比明显增加(P<0.05);与正常组比,NS组仅BALF中的MCP-1水平、W/D比在3 h时明显升高(P<0.05),余时间点各检测指标差异均无统计学意义(P>0.05)。结论:可采用本研究方法对小鼠暴露式气管注射水溶性药物。当研究8 h内的肺脏变化时对照组宜予等量0.9%氯化钠溶液;≥8 h时,可以设正常组为对照组。
Objective： To make water-soluble drugs instilled through the exposed trachea a stable and roughly equal distribution in the lungs of mice and to design the control group scientifically. Methods： Intervention measure, mouse position, instillment speed, minimum volume that requires and maximum volume that tolerates were all taken into consideration to make the distribution of the drug （Methlene Blue instead） in lung lobes in mice （weighting from 18 g to 30 g） stable and roughly equal. Then taking lipopolysaccharide （LPS） as example, the LPS group and normal saline （NS） group were treated with LPS （2 mg/mL） and normal saline （NS） respectively in the way above mentioned in mice weighting from 20 g to 24 g. The normal group was with no pretreatment. After modeling for 3 h, 8 h, 14 h, the level of monocyte chemoattractant protein-1 （MCP-1） in bronehoalveolar lavage fluid （BALF） detected by enzyme-linked immunoabsorbent assay （ELISA）, the polymorphonuclear neutrophil percentage （PMN%） in BALF tested with Wright-Giemsa stain, histopathologieal changes assayed by HE staining, the expression of intercellular adhesion molecule 1 （ICAM-1） assayed by immunohistochemical staining and the lung wet weight/dry weight ratio （W/D ratio） in lung tissue would be tested. Results： Using the process developed above, the distribution of Methlene Blue in lung lobes was stable and roughly equal. Compared with the normal group, the level of MCP-1 in BALF in the LPS group increased at 3 h, 8 h, and as time went by, the PMN% in BALF increased and histopathological changes appeared obvious, and both the expression level of ICAM-1 and the W/D ratio increased （P〈0.05） in LPS group. Compared with the normal group, only the level of MCP-1 in BALF and the W/D ratio increased significantly （P〈0.05） in the NS group at 3 h, and all measurements had no difference at 8 h and 14 h. Conclusion： The process developed here to instill watersoluble drugs through exposed trachea is successful. The control group should be treated with NS for the same time if researching the lung changes within 8 h, or just the normal mice instead if at least 8 hours.
作者
王怒静
彭传鹏
郑爽
周玲萍
徐红蕾
WANG Nujing PENG Chuanpeng ZHENG Shuang ZHOU Lingping XU Honglei(The First Clinical Medical College, Wenzhou Medical University, Wenzhou, 325035 Department of Respiratory Medicine, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325015)
出处
《温州医科大学学报》
CAS
2017年第6期401-405,共5页
Journal of Wenzhou Medical University
基金
温州市科技局科研基金资助项目(Y20140680)
浙江省卫生厅科研基金资助项目(2015RCB018)
浙江省自然科学基金资助项目(LY12H01001)
关键词
小鼠
气管注射
水溶性药物
对照组
mice
intratracheal instillation
water-soluble drug
control group
