摘要
目的探究BPOZ-2在缓解LPS诱导的小鼠急性肺损伤(ALI)中的作用。方法C57BL/6J小鼠气管内滴注不同剂量LPS(1、2.5和5 mg/kg)作为实验组,同时以PBS作为对照组,构建小鼠ALI模型。收集支气管肺泡灌洗液(BALF),利用流式细胞仪分析BALF中的细胞数量;通过BCA蛋白定量法检测其中的总蛋白水平;利用ELISA分析BALF中的炎症因子水平;取小鼠肺组织进行HE染色,分析小鼠肺组织的损伤程度,采用RT-qPCR技术检测小鼠肺组织中bpoz-2的转录水平;Western blot和免疫组化法检测小鼠肺组织中BPOZ-2的蛋白表达水平。在C57BL/6J小鼠中分别气管内滴注7.0×10^(10)v.g AAV-bpoz-2,并以AAV-GFP为对照,3~4周后,利用RT-qPCR分析小鼠肺组织中bpoz-2的转录水平。待BPOZ-2过表达后,按照5 mg/kg的剂量气管内滴注LPS,72 h后,收集BALF,取小鼠肺组织,分析小鼠ALI的程度。结果LPS诱导的小鼠ALI程度随LPS剂量的增加而加重,BALF中的蛋白浓度、细胞数量以及炎症因子TNF-α、IL-6显著增加;小鼠肺组织中BPOZ-2的转录及蛋白表达水平随着LPS剂量的增加以及诱导时间的延长显著降低;过表达BPOZ-2后,小鼠肺组织表观观察以及HE染色的结果显示其损伤面积和程度明显减弱;BALF中的蛋白浓度、细胞数量以及炎症因子TNF-α、IL-6、IL-1β均显著降低,LPS诱导的小鼠ALI相关症状减轻。结论过表达BPOZ-2可以缓解LPS诱导的小鼠ALI。
This study was designed to explore the role of BPOZ-2 in alleviating LPS-induced acute lung injury(ALI)in mice.C57BL/6J mice were given different doses of LPS(1,2.5 and 5 mg/kg)or PBS by intratracheal instillation to establish mouse ALI model or controls.Bronchoalveolar lavage fluid(BALF)was collected and flow cytometry was used to analyze the number of cells;total proteins in BALF were detected by BCA protein quantitative method,and cytokines in BALF were analyzed by ELISA.HE staining was used to analyze the degree of lung injury.The transcriptional level of bpoz-2 in lung tissues of mice were detected by RT-qPCR,and the level of protein expression was detected by Western blotting and immunohistochemistry.C57BL/6J mice were given 7.0×10^(10) v.g AAV-BPOZ-2 by intratracheal instillation,while AAV-GFP was used as control.And 3-4 weeks later,RT-qPCR was used to analyze the transcriptional level of bpoz-2 in lung tissues of mice.After overexpression of BPOZ-2,mice were given 5 mg/kg LPS by intratracheal instillation for 72 h,and then BALF and the lung tissues were collected to analyze the degree of ALI.Data showed that the ALI induced by LPS in mice was significantly aggravated with the increase of dose of LPS.The protein concentrations,number of cells and cytokines such as TNF-αand IL-6 in BALF were increased significantly,while the transcription level and protein expression of BPOZ-2 in lung tissues decreased significantly with the increase of LPS dose and induction time.After overexpression of BPOZ-2,the lung injury area and degree decreased significantly,the protein concentrations,number of cells and cytokines such as TNF-α,IL-6 and IL-1βin BALF were all significantly decreased,and the symptoms related to ALI induced by LPS were alleviated.Taken together,overexpression of BPOZ-2 can alleviate LPS-induced ALI in mice.
作者
范廷辉
林浩天
杨小盼
钟辉
王炜
王斌
FAN Tinghui;LIN Haotian;YANG Xiaopan;ZHONG Hui;WANG Wei;WANG Bin(School of Basic Medicine,Qingdao University,Qingdao 266071,China;Institute of Biotechnology,Academy of Military Medical Sciences,Beijing 100071,China;Health Management Center,Zhuzhou Center Hospital,Zhuzhou 412007,China)
出处
《免疫学杂志》
CAS
CSCD
北大核心
2023年第9期746-753,760,共9页
Immunological Journal
基金
中国博士后科学基金(2020M683743)。
