阿尔法骨化醇联合地塞米松减轻小鼠的肺纤维化及其机制

Alphacalcidol combined with dexamethasone for reducing pulmonary fibrosis in mice and its mechanism

目的研究阿尔法骨化醇联合地塞米松对肺纤维化影响。方法 C57BL/6小鼠40只,随机分为正常对照组、肺纤维化组、阿尔法骨化醇组、地塞米松组、地塞米松联合阿尔法骨化醇组。正常对照组予以0.5μL/g生理盐水一次性气管注射,同时每天胃内灌注生理盐水0.2 m L;其他组予以0.5μL/g博来霉素溶液一次性气管注射,肺纤维化组每天给予生理盐水0.2 m L灌胃,地塞米松组予以地塞米松[0.5 mg/(kg·d)]灌胃,阿尔法骨化醇组予以阿尔法骨化醇[0.5μg/(kg·d)]灌胃,地塞米松联合阿尔法骨化醇组给予地塞米松[0.5 mg/(kg·d)]联合阿尔法骨化醇[0.5μg/(kg·d)]灌胃,21 d后处死。行HE及Masson染色观察肺组织病理变化,收集血清检测各组羟脯氨酸(Hyp)值,收集支气管肺泡灌洗液(BALF),沉淀重悬行细胞计数,上清液行ELISA检测白细胞介素1β(IL-1β)、IL-6、肿瘤坏死因子α(TNF-α)及转化生长因子β1(TGF-β1)。结果阿尔法骨化醇联合地塞米松组IL-1β、IL-6、TNF-α及TGF-β1,细胞计数、Sazpiel评分、Hyp值均低于除对照组外其他各组,其肺泡炎情况及纤维沉积也均低于除对照组外其他各组。结论阿尔法骨化醇联合地塞米松对肺纤维疗效优于地塞米松及阿尔法骨化醇单药疗效,其机制可能与抑制TGF-β1、炎性因子相关。

Objective To study the effect of alphacalcidol and dexamethasone on pulmonary fibrosis and its mechanism. Methods Forty C57BL/6 mice were randomly divided into five groups： normal control group, pulmonary fibrosis group, alphacalcidol group, dexamethasone group, dexamethasone combined with alphacalcidol group. The normal control group was given 0. 5 μL/g normal saline by one-time intratracheal injection as well as 0. 2 mL normal saline by daily intragastric infusion. The other groups received 0. 5 μL/g bleomycin solution by one-time intratracheal injection; besides, the pulmonary fibrosis group were subjected to 0.2 mL normal saline gavage; the dexamethasone group were treated with dexamethasone [0.5 mg/ （kg · d）] orally; the alphacalcidol group received intragastric infusion of alphacalcidol [0.5 μg/ （kg · d）]; and the dexamethasone plus alphacalcidol group was given dexamethasone [ 0.5 mg/ （kg · d ）] mixed with alphacalcidol [0.5 μg/ （ kg · d） ] orally. Then they were killed after 21 days. The pathological changes of lung tissues was observed using HE and Masson staining. The serum was collected to detect hydroxyproline （Hyp） values. Bronchoalveolar lavage fluid （BALF） was collected for cell counting. ELISA was performed to examine interleukin 1 beta （IL-1β）, IL-6, tumor necrosis factor alpha （TNF-α） and transforming growth factor beta 1 （TGF-IBI） in supernatants. Results In the alphacalcidol combined with dexamethasone group, IL-1β, IL-6, TNF-α and TGF-β1, cell count, Sazpiel score, Hyp values were lower than those in the other groups except for the control group, and the alveolar inflammation and fibrin deposition were also lower than those in the other groups except for the control group. Conclusion The curative effect of alphacalcidol combined with dexamethasone on pulmonary fibrosis is better than either dexamethasone or alphacalcidol alone, and the mechanism may be related to the inhibition of TGF-β1 and inflammatory factors.