摘要
目的探讨人脐带间充质干细胞来源外泌体(hUCMSC-Exo)对大鼠放射性肺损伤(RILI)的作用及机制。方法流式细胞术检测人脐带间充质干细胞(hUC-MSC)表面标志物;蛋白免疫印迹法、纳米粒子跟踪分析及透射电子显微镜鉴定hUCMSC-Exo。根据随机数字表法将15只健康SD大鼠分为对照组、照射组和照射+Exo组,每组各5只,照射+Exo组大鼠在辐照后立即注射100μl的hUCMSC-Exo(剂量为1×10^(7)个/μl),其余两组注射100μl生理盐水,照射后4周处死大鼠并取材;苏木精-伊红(HE)和Masson染色法观察肺组织病理学变化;酶联免疫吸附法(ELISA)检测大鼠血清中白细胞介素1β(IL-1β)、白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)和干扰素γ(INF-γ)表达;蛋白免疫印迹法检测大鼠右肺组织中E-钙黏蛋白(E-cadherin)、波形蛋白(vimentin)、转化生长因子-β1(TGF-β1)和IL-1β蛋白表达;实时荧光定量聚合酶链式反应(PCR)检测E-cadherin、Vimentin、IL-1β和IL-6的mRNA表达;免疫组织化学染色检测大鼠右肺组织中E-cadherin、Vimentin、TGF-β1和磷酸化Smad同源物(p-Smad)蛋白表达。结果与照射组相比较,照射+Exo组炎性细胞浸润减少,胶原沉积和纤维组织增生减少(P<0.05);血清中促炎因子IL-1β、IL-6、TNF-α及INF-γ表达减少(P<0.05);肺组织中E-cadherin蛋白表达增加,Vimentin、TGF-β1、p-Smad和IL-1β蛋白表达减少(P<0.05);肺组织中E-cadherin的mRNA表达增加,Vimentin、IL-1β及IL-6的mRNA表达减少(P<0.05)。结论hUCMSC-Exo能够通过减少促炎因子分泌及抑制纤维化进展缓解RILI。
Objective To investigate the alleviating effect and mechanism of human umbilical cord mesenchymal stem cells-derived exosome(hUCMSC-Exo)on radiation induced lung injury(RILI)in rats.Methods The human umbilical cord mesenchymal stem cells(hUC-MSC)surface markers were detected by flow cytometry.The western blot,nanoparticle tracking analysis and transmission electron microscopy were used to identify hUCMSC-Exo.The 15 healthy SD rats were randomly divided into 3 groups:the control group,the irradiation group and the irradiation+Exo group,with 5 rats in each group.The rats in the irradiation+Exo group were injected with 100μl hUCMSC-Exo once immediately after irradiation(the dose was 1×10^(7) particles/μl),and the rats in the other 2 groups were injected with 100μl normal saline and were killed 4 weeks after irradiation and samples were collected.The pathological changes of lung were observed by hematoxylin-eosin(HE)and Masson staining.The expression of inflammatory factors interleukin-1β(IL-1β),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α)and interferon-γ(INF-γ)in serum of rats was detected by enzyme-linked immunosorbent assay(ELISA).The expressions of E-cadherin,Vimentin,transforming growth factor-β1(TGF-β1)and IL-1βin the right lung were detected by western blot.The mRNA expression of E-cadherin,Vimentin,IL-1βand IL-6 was detected by real-time fluorescence quantitative polymerase chain reaction(PCR).The expressions of E-cadherin,Vimentin,TGF-β1 and phosphorylation mothers against decapentaplegic homolog(p-Smad)in the right lung were detected by immunohistochemistry.Results Compared with the irradiation group,the inflammatory cell infiltration,collagen deposition and fibrous tissue hyperplasia were decreased in the lung tissue of the irradiation+Exo group(P<0.05).Serum expressions of pro-inflammatory factors IL-1β,IL-6,TNF-αand INF-γwere decreased(P<0.05).The expression of E-cadherin protein was increased,and the expression of Vimentin,TGF-β1,p-Smad and IL-1βprotein was decreased(P<0.05).Th
作者
杨紫恩
欧阳明玥
王丽丽
刘硕
孙凌燕
阎英
刘宇
于卉影
YANG Zi-en;OUYANG Ming-yue;WANG Li-li;LIU Shuo;SUN Ling-yan;YAN Ying;YU Hui-ying(Department of Basic Medical Laboratory,General Hospital of Northern Theater Command,Shenyang 110016,China)
出处
《创伤与急危重病医学》
2024年第4期216-221,226,共7页
Trauma and Critical Care Medicine
关键词
人脐带间充质干细胞来源外泌体
放射性肺损伤
促炎因子
纤维化
Human umbilical cord mesenchymal stem cells-derived exosome
Radiation-induced lung injury
Proinflammatory factors
Fibrosis
