摘要
目的探讨微小RNA-199a-3p(miR-199a-3p)通过沉默信息调节因子1(SIRT1)/核因子-κB(NF-κB)通路抑制支原体肺炎发展的作用。方法80只SPF级BALB/c小鼠随机分为对照组、模型组、miR-199a-3p组及抑制组,每组各20例。除对照组外,通过高载量肺炎支原体菌液连续滴鼻3 d建立支原体肺炎小鼠模型。造模后,各组小鼠均进行尾静脉注射,miR-199a-3p组和抑制组小鼠分别注射agomir液、antagomir液,对照组和模型组小鼠均给予尾静脉注射等量生理盐水。实验结束后处死各组小鼠,各组小鼠均眼球取血,酶联免疫吸附试验检测白细胞介素-4(IL-4)、白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)水平。取各组小鼠肺组织,行HE染色观察肺组织病理改变,实时荧光定量PCR法检测肺组织miR-199a-3p基因表达水平,Western Blot检测肺组织SIRT1/NF-κB信号通路蛋白表达。结果与模型组比较,miR-199a-3p组血清IL-4、IL-6、IL-1β和TNF-α水平均降低,差异均有统计学意义(P<0.05)。HE染色结果显示,对照组小鼠的肺组织结构基本正常且无肺泡内渗出或炎性反应,其余3组均存在不同程度的肺泡间质增宽、肺泡内渗出以及炎性细胞浸润。与模型组比较,miR-199a-3p组miR-199a-3p基因表达水平和SIRT1蛋白相对表达水平升高,差异均有统计学意义(均P<0.05);NF-κB蛋白相对表达水平降低,差异有统计学意义(P<0.05)。结论miR-199a-3p基因在支原体肺炎小鼠肺组织中表达降低,提高miR-199a-3p基因表达水平通过抗炎作用对支原体肺炎小鼠肺组织损伤起到保护作用,其作用机制可能与其对SIRT1/NF-κB通路的调控有关。
Objective To explore the effects of MiR-199a-3p on promoting the development of mycoplasma pneumonia via the silent information regulator 1(SIRT1)/nuclear factor-κB(NF-κB)pathway.Methods Totally 80 SPF-grade BALB/c mice were randomly divided into a control group,a model group,a miR-199a-3p group,and a inhibitory group,with 20 rats in each group.Excep the control group,the model group was established as a mouse model of mycoplasma pneumoniae through a continuous nasal drip of a high-load mycoplasma pneumoniae bacterial solution for 3 days.After modeling,mice in each group had tail vein injections.The miR-199a-3p group and the inhibitory group mice were injected with agomir solution and antagonir solution,respectively.And the model group and control group mice were injected with the same amount of physiological saline through the tail vein.After the experiment,mice of all groups were killed and their blood was collected from the eyeballs,and interleukin-4(IL-4),interleukin-6(IL-6),interleukin-1β(IL-1β),and tumor necrosis factor-α(TNF-α)levels were detected by the enzyme-linked immunosorbent assay method.Subsequently,the lung tissues of the mice were taken for HE staining to observe pathological changes in the lung tissue.Real-time fluorescence quantitative PCR was used to detect miR-199a-3p gene expression levels in lung tissue,and Western Blot was used to detect SIRT1/NF-κB signaling pathway protein expression in lung tissue.Results Compared with the model group,IL-4,IL-6,IL-1β,and TNF-αlevels in the serum of the miR-199a-3p group were decreased,with a significant difference(P<0.05).The HE staining results showed that the lung tissue structure of the control group mice was nearly normal and there was no alveolar exudation or inflammatory response.The other three groups all had varying degrees of alveolar interstitial widening,alveolar exudation,and inflammatory cell infiltration.Compared with the model group,miR-199a-3p gene and SIRT1 protein expression levels in the miR-199a-3p group increased,with a sig
作者
赵鲁笑
冯战超
高继勇
Zhao Luxiao;Feng Zhanchao;Gao Jiyong(Department of Pediatrics,Jinan Maternal and Child Health Hospital,Jinan 250001,China;Department of Children’s Health,Danxian Maternal and Child Health Hospital,Heze 274300,China;Rehabilitation Center of Children’s Medical,Jinan Maternal and Child Health Hospital,Jinan 250001,China)
出处
《国际生物医学工程杂志》
CAS
2024年第2期167-173,共7页
International Journal of Biomedical Engineering
