甲泼尼龙联合环磷酰胺对博来霉素诱导的肺纤维化大鼠模型炎症反应与免疫细胞活性的影响  

Effects of methylprednisolone combined with cyclophosphamide on inflammatory reaction and immune cell activity in bleomycin-induced pulmonary fibrosis rat model

作　　者：徐庆杰 陈燕 张曼卡 李周平 刘音[1] 唐铭 饶芝国[1] 马润林 薛晓艳[1] Xu Qingjie;Chen Yan;Zhang Manka;Li Zhouping;Liu Yin;Tang Ming;Rao Zhiguo;Ma Runlin;Xue Xiaoyan(Department of Critical Care Medicine,Aerospace Center Hospital,Beijing 100049,China;Department of Genetics and Developmental Biology,College of Life Sciences,University of Chinese Academy of Sciences,Beijing 100049,China)

机构地区：[1]航天中心医院重症医学科,北京100049 [2]中国科学院大学生命科学学院遗传与发育生物学教研室,北京100049

出　　处：《药物不良反应杂志》2023年第12期732-738,共7页Adverse Drug Reactions Journal

基　　金：分子发育生物学国家重点实验室开放项目(2021-MDB-KF-13);航天医科科研基金(2020YK03)。

摘　　要：目的观察甲泼尼龙(MP)联合环磷酰胺(CTX)对博来霉素(BLM)诱导的肺纤维化大鼠模型炎症和免疫细胞活性的影响。方法将健康6~8周龄SD大鼠40只以随机数字表法分为空白对照、BLM模型、BLM+MP和BLM+MP+CTX组,每组10只。采用气管内灌注BLM(一次性给予5 mg/kg)的方法制备肺纤维化大鼠模型,造模第7天开始每天予BLM+MP组尾静脉注射MP(3 mg/kg),BLM+MP+CTX组注射MP(3 mg/kg)+CTX(8 mg/kg),持续给药21 d。采用苏木精-伊红(HE)和Masson染色检测大鼠肺部炎症及纤维化程度;采用流式细胞术检测大鼠支气管肺泡灌洗液(BALF)中粒细胞、中性粒细胞数量及血液T细胞亚群。结果造模第7天,大鼠肺组织外部形态、HE及Masson染色结果显示BLM诱导肺纤维化模型制备成功。造模第28天,BLM组大鼠肺组织结构紊乱且有明显胶原沉积,BALF中粒细胞和中性粒细胞数量显著增多,血液T细胞、CD4^(+)T细胞和调节性T细胞(Tregs)占比下降,CD8^(+)T细胞占比增高,CD4^(+)/CD8^(+)T细胞(CD4/CD8)比值降低(均P<0.05)。与BLM组相比,BLM+MP+CTX组大鼠肺纤维化程度得到明显改善,BALF中粒细胞和中性粒细胞数量显著减少,血液T细胞、CD4^(+)T细胞和Tregs细胞占比升高,CD8^(+)T细胞占比降低,CD4^(+)/CD8^(+)T细胞(CD4/CD8)比值增高(均P<0.05)。BLM+MP+CTX组的改善情况优于BLM+MP组,差异有统计学意义(P<0.05)。结论MP联合CTX可减轻肺纤维化大鼠的炎症反应程度,提高T细胞免疫活性。Objective To observe the effect of methylprednisolone(MP)combined with cyclophosphamide(CTX)on inflammation and immune cell activity in bleomycin(BLM)-induced pulmonary fibrosis rat model.Methods Forty healthy 6 to 8-week-old SD rats were randomly divided into blank control,BLM model,BLM+MP,and BLM+MP+CTX groups,with 10 rats in each group.The rat model of pulmonary fibrosis was prepared by intratracheal infusion of BLM(5-mg/kg,only once).From the 7th day of modeling,MP(3-mg/kg)was injected in rats in the BLM+MP group and MP(3-mg/kg)+CTX(8-mg/kg)was injected via tail vein in rats in the BLM+MP+CTX group,once daily for 21 days.The degree of lung inflammation and fibrosis in rats was detected using HE and Masson staining methods.The numbers of granu-locytes and neutrophils in bronchoalveolar lavage fluid(BALF)and blood T cell subsets in rats were detected using flow cytometry.Results On the 7th day of modeling,the external morphology,HE and Masson staining results of rat lung tissue showed that BLM-induced pulmonary fibrosis model was successfully prepared.On the 28th day of modeling,the lung tissue structure of the BLM group was disordered with obvious collagen deposition,the number of granulocytes and neutrophils in BALF increased significantly,the proportion of blood T cells,CD4^(+)T cells,and regulatory T cells(Tregs)decreased,the proportion of CD8^(+)T cells,and the CD4^(+)/CD8^(+)T cells ratio decreased significantly(all P<0.05).Compared with the BLM group,the degree of pulmonary fibrosis in the BLM+MP+CTX group was improved significantly,the number of granulocytes and neutrophils in BALF decreased significantly,the proportion of blood T cells,CD4^(+)T cells and Tregs cells increased significantly,the proportion of CD8^(+)T cells decreased,and the ratio of CD4^(+)/CD8^(+)T cells increased significantly(all P<0.05).The improvement effect in rats of BLM+MP+CTX group was better than that of BLM+MP group,and the difference was statistically significant(P<0.05).Conclusion MP combined with CTX can reduce the degree

关 键 词：肺纤维化 疾病模型 动物 炎症 甲泼尼龙 环磷酰胺 T淋巴细胞 免疫活性 

分 类 号：R563[医药卫生—呼吸系统] R-332[医药卫生—内科学]