维生素D干预对支气管哮喘幼鼠外周血T淋巴细胞平衡调节及肠道菌群的影响  

Effects of vitamin D intervention on balance regulation of peripheral blood T lymphocytes and intestinal flora in young rats with bronchial asthma

作　　者：蔡姿秀 周国华[1] 田军 CAI Zixiu;ZHOU Guohua;TIAN Jun(Department of Pediatrics,the First People’s Hospital of Xiaoshan District,Hangzhou City,Zhejiang Province,Hangzhou 311200,China;Department of Pediatrics,the Third People’s Hospital of Xiaoshan District,Hangzhou City,Zhejiang Province,Hangzhou 311256,China)

机构地区：[1]杭州市萧山区第一人民医院儿科,浙江杭州311200 [2]杭州市萧山区第三人民医院儿科,浙江杭州311256

出　　处：《中国医药导报》2023年第16期21-25,36,共6页China Medical Herald

基　　金：浙江省医药卫生科技计划项目(2022ZH043);浙江省杭州市萧山区重大科技计划项目(2020301)。

摘　　要：目的研究维生素D(VitD)干预对支气管哮喘(以下简称“哮喘”)幼鼠外周血T淋巴细胞平衡调节及肠道菌群的影响。方法选取30只7 d龄SPF级野生型BALB/c雌性小鼠(体重4.09~6.13 g)为研究对象,采用随机数字表法将其分为哮喘模型组(15只)和VitD干预组(15只),另选取10只为健康对照组。实验第1、7天,哮喘模型组及VitD干预组采用10%卵清蛋白(OVA)致敏液200μl腹腔注射,健康对照组注射等量生理盐水。第18天,VitD干预组给予VitD滴剂[35μg/(kg·周),1次/2 d]灌胃,共给药2周,健康对照组、哮喘模型组灌胃等量磷酸盐缓冲液。第19天,哮喘模型组和VitD干预组给予3%OVA溶液10 ml雾化吸入20 min,健康对照组给予等量生理盐水雾化吸入,1次/d,共6 d。实验结束后,观察三组肺组织切片苏木精-伊红染色结果;采用流式细胞仪检测并计算三组血清Th1/Th2、Th17/调节性T细胞(Treg)比值;通过16S rDNA测序法分析三组粪便内菌群情况。结果健康对照组肺组织支气管壁及肺泡壁结构完整,肺泡腔内无渗出,肺泡上皮排列有序,肺泡壁厚度正常;哮喘模型组肺泡壁结构受损,肺泡上皮细胞破坏,黏膜充血水肿,血管周围炎症细胞浸润;VitD干预组肺组织炎症反应较轻。三组测序样本微生物物种Shannon指数曲线趋平缓,三组Simpson指数比较,差异无统计学意义(P>0.05)。PCoA图分析显示,哮喘模型组和健康对照组组间距离大,VitD干预组与哮喘模型组组间距离近(P<0.05)。三组肠道菌群涉及15个门,23个属。哮喘模型组Th1/Th2比值及Actinobacteriota、Helicobacter、Colidextribacter丰度低于健康对照组,Th17/Treg比值及Proteobacteria、Ligilactobacillus、Bacteroides丰度高于健康对照组(P<0.05)。VitD干预组Thl/Th2比值及Actinobacteriota、Proteobacteria、Helicobacter、Colidextribacter丰度高于哮喘模型组,Th17/Treg比值及Ligilactobacillus、Bacteroides丰度低于哮喘模型组(P<0.05),与基于属水平的Objective To investigate the effects of vitamin D(VitD)intervention on the balance regulation of peripheral blood T lymphocytes and intestinal flora in young rats with bronchial asthma(hereinafter referred to as“asthma”).Methods Thirty SPF wild-type BALB/c female mice(4.09-6.13 g)aged seven days were selected as the study objects,and they were divided into asthma model group(15 rats)and VitD intervention group(15 rats)by random number table method,another ten rats were selected as healthy control group.On the first and seventh day of the experiment,asthma model group and VitD intervention group were injected intraperitoneally with 10%ovalbumin(OVA)sensitizing solution 200μl,while healthy control group was injected with the same amount of normal saline.On the 18th day,VitD intervention group was given VitD drops(35μg/kg,per week,one every two days)for two weeks,and healthy control group and asthma model group were given the same amount of phosphate buffer.On the 19th day,asthma model group and VitD intervention group were given atomized inhalation of 3%OVA solution 10 ml for 20 min,while healthy control group was given atomized inhalation of normal saline,once a day for six days.After the experiment,the hematoxylin-eosin staining results of lung tissue sections in three groups were observed.Serum Th1/Th2 and Th17/regulatory T cell(Treg)ratios were measured and calculated by flow cytometry.The bacterial community in feces of the three groups was analyzed by 16S rDNA sequencing method.Results In the healthy control group,the structure of the bronchial wall and alveolar wall of the lung tissue were complete,there was no exudation in the alveolar cavity,the alveolar epithelium was orderly,and the alveolar wall thickness was normal.In asthmatic model group,the structure of alveolar wall was damaged,alveolar epithelial cells were destroyed,mucosal hyperemia and edema were observed,and perivascular inflammatory cells were infiltrated.The inflammatory reaction of lung tissue in VitD intervention group was less.The S

关 键 词：维生素D 支气管哮喘 肠道菌群 外周血T淋巴细胞平衡 调节机制 

分 类 号：R725.6[医药卫生—儿科]