急性肺损伤对大鼠脾T细胞凋亡的影响及益肺健脾方的干预作用研究  

Effects of acute lung injury on apoptosis in rat splenic T cells and the interventional effect of Yifei Jianpi formula

作　　者：杨玉洁 朱中博 张艳美 刘喜平[1] 张旭辉 张志明 YANG Yujie;ZHU Zhongbo;ZHANG Yanmei;LIU Xiping;ZHANG Xuhui;ZHANG Zhiming(Gansu University of Chinese Medicine,Laboratory for TCM New Products Development Engineering of Gansu Province,Key Laboratory of Traditional Chinese Herbs and Prescription Innovation and Transformation of Gansu Province,Lanzhou 730000,China;Department of Pulmonary Disease,Affiliated Hospital of Gansu University of Chinese Medicine,Lanzhou 730000,China;Gansu Provincial Hospital of Traditional Chinese Medicine,Lanzhou 730000,China)

机构地区：[1]甘肃中医药大学,甘肃省中药新产品创制工程实验室,甘肃省中医方药挖掘与创新转化重点实验室,兰州730000 [2]甘肃中医药大学附属医院,肺病科,兰州730000 [3]甘肃省中医院,兰州730000

出　　处：《中国实验动物学报》2024年第9期1160-1170,共11页Acta Laboratorium Animalis Scientia Sinica

基　　金：甘肃省科技重大项目(22ZD1FA001);国家自然科学基金(82260889);第五批全国中医临床优秀人才研修项目(国中医药人教函[2022]239号);甘肃省自然科学基金项目(24JRRD002)。

摘　　要：目的观察急性肺损伤(acute lung injury,ALI)大鼠脾T细胞凋亡及X-连锁凋亡抑制蛋白相关因子(XIAP-associated factor 1,XAF1)、肿瘤坏死因子α(tumor necrosis factor-alpha,TNF-α)、死亡相关因子(factor associatedsuicide,FAS)蛋白表达情况,探讨益肺健脾方治疗ALI的机制是否与下调XAF1、FAS、TNF-α蛋白表达,抑制T细胞凋亡有关。方法60只SPF级雄性SD大鼠,随机分为空白组、模型组、阳性组、益肺健脾方高、中、低剂量组。阳性组给予地塞米松每天0.5 g/kg灌胃,益肺健脾方高、中、低剂量组分别给予每天12、6、3 g/kg的益肺健脾方灌胃,模型组与空白组均给予等量蒸馏水灌胃,每天给药1次,连续14 d。动物肺功能检测系统检测各组大鼠肺功能情况;观察各组大鼠肺部影像学特征及脏器指数和肺组织湿重/干重(W/D)变化;苏木素-伊红(HE)染色观察各组大鼠肺部组织的病理学变化;流式细胞术检测各组大鼠脾T细胞亚群(CD4^(+)/CD8^(+))及脾T细胞的凋亡情况,Western Blot检测脾中XAF1、FAS、TNF-α蛋白表达水平。结果模型组大鼠肺功能降低,脾和胸腺脏器指数减少,肺组织湿重/干重(W/D)显著升高(P<0.01),肺部组织出现炎性渗出、肺泡破裂等现象,同时伴有肺纹理增粗以及大片磨玻璃影,T细胞亚群(CD4^(+)/CD8^(+))显著减少,XAF1、FAS、TNF-α蛋白表达及T细胞凋亡率显著升高(P<0.01);经益肺健脾方干预后,大鼠肺组织湿重/干重(W/D)明显降低,胸腺的脏器指数显著升高(P<0.05,P<0.01),T细胞亚群(CD4^(+)/CD8^(+))明显增加,XAF1、FAS、TNF-α蛋白表达及T细胞凋亡率明显下降(P<0.05,P<0.01)。结论ALI可诱发大鼠脾XAF1、FAS、TNF-α蛋白表达上调及T细胞凋亡,益肺健脾方可能通过下调XAF1、FAS、TNF-α蛋白表达,抑制ALI诱发的脾T细胞凋亡。Objective To observe splenic T cell apoptosis and XIAP-associated factor 1(XAF1),FAS,and tumor necrosis factor(TNF)-αprotein expression levels in rats with acute lung injury(ALI),and to determine their roles in the protective effect of Yifei Jianpi formula.Methods Sixty male specific pathogen-free SD rats were divided randomly into blank,model,positive control,and high-,medium-,and low-dose Yifei Jianpi formula groups.Rats in the positive control group were given 0.5 g/kg dexamethasone by gavage,and rats in the high-,medium-,and low-dose Yifei Jianpi formula groups were given 12,6,and 3 g/kg Yifei Jianpi formula by gavage,respectively.Rats in the model and blank groups were given equal amounts of saline by gavage.All medications were administered once a day for 14 days.Lung function testing was carried out in all rats.We observed the imaging characteristics of the lungs and changes in the organ index and lung tissue wet/dry weight(W/D)in each group,and detected the pathological changes in lung tissues by hematoxylin-eosin staining.Splenic T-cell subpopulations(CD4^(+)/CD8^(+))and apoptosis of splenic T-cells were detected by flow cytometry and XAF1,FAS,and TNF-αprotein expression levels in the spleen were detected by Western Blot.Results Rats in the model group showed reduced lung function,decreased spleen and thymus organ indexes,and significantly higher W/D of lung tissue(P<0.01).In addition,they had inflammatory exudation and alveolar rupture in the lung tissue,accompanied by thickening of the lung texture and large areas of ground-glass shadows,with a significant decrease in T-cell subsets(CD4^(+)/CD8^(+))and significant increases in XAF1,FAS,and TNF-αproteins,and in the rate of T-cell apoptosis(P<0.01).Yifei Jianpi formula significantly reduced the W/D spleen of rat lung tissues,significantly increased the thymus organ index(P<0.05,P<0.01)and the T-cell subpopulation(CD4^(+)/CD8^(+)),and significantly decreased the protein expression levels of XAF1,FAS,and TNF-α,and the T-cell apoptosis rate(P<0.05,P<0.0

关 键 词：急性肺损伤 益肺健脾方 T细胞凋亡 XAF1 FAS TNF-Α 

分 类 号：Q95-33[生物学—动物学]