摘要
目的探讨衰老相关肺纤维化与免疫功能的关系及细菌溶解产物对老年小鼠肺纤维化的影响。方法24只老年健康雄性C57BL/6小鼠随机分为对照组(N组)、肺纤维化组(M组)、肺纤维化+细菌溶解产物干预组(P组),M、P组于气管内注射博来霉素(5 mg/kg)建立小鼠肺纤维化模型,N组注射生理盐水,P组建模后经口注入细菌溶解产物0.4 ml,1次/d,造模28 d后取肺组织和血液。通过苏木精-伊红(HE)和马松三色(Masson)染色检测小鼠肺组织病理变化并进行Ashcroft评分,免疫组化检测肺组织CD4+、CD8+表达,酶联免疫吸附测定(ELISA)法检测小鼠血清干扰素-γ(INF-γ)、白细胞介素-13(IL-13)和免疫球蛋白A(IgA)蛋白含量,实时定量反转聚合酶链式反应(RT-qPCR)检测肺组织INF-γ、IL-13 mRNA的含量,蛋白质免疫印迹法(Western Blot)检测肺组织基质金属蛋白酶-9(MMP-9)和组织型金属蛋白酶抑制剂-1(TIMP-1)蛋白表达水平。结果P组小鼠在细菌溶解产物干预下肺纤维化程度较M组明显减轻;与N组比较,M组CD4+T细胞表达明显减少,CD8+T细胞表达明显增加(P<0.05),IgA含量减少(P<0.05);与M组比较,P组CD4+T细胞表达明显增加,CD8+T细胞表达明显减少(P<0.05),IgA含量升高(P<0.05);细菌溶解产物干预后,IFN-γ的mRNA和蛋白表达水平升高(P<0.05),而IL-13的mRNA和蛋白表达水平降低(P<0.05);与N组相比,M组MMP-9和TIMP-1的蛋白表达显著上调(P<0.05),细菌溶解产物干预后MMP-9和TIMP-1蛋白表达降低(P<0.05)。结论免疫机制参与了肺纤维化的形成,细菌溶解产物可以通过调节机体的免疫平衡状态,从而在一定程度上减轻博来霉素诱导的老年小鼠肺纤维化。
Objective To investigate the correlation between immune function and age-related pulmonary fibrosis,as well as the potential impact of bacterial lysates on this condition.MethodsTwenty-four healthy male C57BL/6 mice,aged 24,were randomly divided into three groups:a control group(Group N),a pulmonary fibrosis group(Group M),and a pulmonary fibrosis+bacterial lysis product intervention group(Group P).Mice in Groups M and P were intratracheally injected with bleomycin(5 mg/kg)to induce a mouse pulmonary fibrosis model,while mice in Group N were injected with saline.After modeling,mice in Group P were orally administered 0.4 ml of a bacterial lysis product once a day.After 28 days,lung tissue and blood samples were collected for analysis.Pathological changes in lung tissue were assessed using hematoxylin and tosin staining(HE)and Masson staining and the Ashcroft score.The expression of CD4+and CD8+in lung tissue was evaluated using immunohistochemistry.The levels of serum interferon-γ(INF-γ),interleukin-3(IL-13),and immunoglobulin A(IgA)protein were measured using Enzyme-linked Immuno Sorbent Assay(ELISA).The levels of INF-γand IL-13 mRNA in lung tissue were determined using Real-Time Quantitative Transcription PCR(RT-qPCR).Additionally,the protein expression levels of matrix metalloprotein-9(MMP-9)and tissue inhibitor of metalloproteincise 1(TIMP-1)in lung tissue were assessed using blot analysis.ResultsThe degree of lung fibrosis was significantly reduced in mice in group P compared with group M when treated with bacterial lysis products.Group M showed a significant decrease in the expression of CD4+T cells and an increase in the expression of CD8+T cells(P<0.05)compared to group N.Additionally,the content of IgA was decreased(P<0.05)in group M.On the other hand,group P showed a significant increase in the expression of CD4+T cells and a decrease in the expression of CD8+T cells(P<0.05)compared to group M.Furthermore,the content of IgA was elevated(P<0.05)in group P.After bacterial lysis product intervention,mRN
作者
钱力
孙梓越
郭香蕊
李丹
刘学军
韩永康
杜毓锋
Qian Li;Sun Ziyue;Guo Xiangrui;Li Dan;Liu Xuejun;Han Yongkang;Du Yufeng(Department of Geriatrics,First Hospital of Shanzi Medical University,Taiyuan 03000l,China;First Clinical Medical College,Shanri Medical University,Taiyuan O3000l,China)
出处
《中华老年医学杂志》
CAS
CSCD
北大核心
2024年第2期209-215,共7页
Chinese Journal of Geriatrics
基金
山西省基础研究计划面上项目(20210302123248)
山西省基础研究计划青年项目(202203021222373)
山西省卫生健康委科研课题(2021051)。
关键词
肺纤维化
免疫
细菌溶解产物
Pulmonary fibrosis
Immanity
Bacterial lysates
