基于AMPK/Sirt1通路探讨赤芍总苷对中性粒细胞性哮喘小鼠气道炎症的影响

Investigating the effect of total paeony glycoside on airway inflammation in mice with neutrophilic asthma based on the AMPK/Sirt1 pathway

关键词:  赤芍总苷  AMPK/Sirt1通路  中性粒细胞  哮喘  气道炎症

Key words:  total paeony glycoside  AMPK/Sirt1 pathway  neutrophils  asthma  airway inflammation

摘要:

[目的] 基于腺苷酸活化蛋白激酶（AMPK）/沉默信息调节因子1（Sirt1）通路探讨赤芍总苷（TPG）对中性粒细胞性哮喘小鼠气道炎症的影响。[方法] 鼻腔滴入卵清蛋白致敏溶液制备哮喘小鼠，造模小鼠随机分为致敏组、TPG低剂量（TPG-低，150 mg/kg TPG）组、TPG高剂量（TPG-高，300 mg/kg TPG）组、地塞米松（4 mg/kg）组、TPG-高+抑制剂（15 mg/kg）组，随后进行雾化激发，对照组小鼠则以生理盐水滴入鼻腔，并采用雾化生理盐水激发，1 h后进行气道阻力分析；收集支气管肺泡灌洗液（BALF），并对细胞分类计数；分析BALF中炎性因子白细胞介素（IL）-6、肿瘤坏死因子-α（TNF-α）、IL-17水平；分离肺组织，检测肺组织病理学及气道黏液变化、AMPK磷酸化（p-AMPK）/AMPK、Sirt1蛋白表达。[结果] 与对照组相比，致敏组呼气间歇值、中性粒细胞等炎性细胞数、IL-6、IL-17、TNF-α水平、炎性评分及高碘酸-希夫（PAS）评分增加，p-AMPK/AMPK、Sirt1表达降低（P<0.05）；与致敏组相比，TPG-低组、TPG-高组、地塞米松组呼气间歇值、中性粒细胞等炎性细胞数、IL-6、IL-17、TNF-α水平、炎性评分及PAS评分降低，p-AMPK/AMPK、Sirt1表达增加（P<0.05），不同剂量TPG间差异有统计学意义（P<0.05）；与TPG-高组相比，TPG-高+抑制剂组呼气间歇值、中性粒细胞等炎性细胞数、IL-6、IL-17、TNF-α水平、炎性评分及PAS评分增加，p-AMPK/AMPK、Sirt1表达降低（P<0.05）。[结论] TPG通过上调AMPK/Sirt1通路减轻中性粒细胞性哮喘小鼠的气道炎症。

Abstract:

[Objective] To investigate the effect of total paeony glycoside（TPG） on airway inflammation in mice with neutrophilic asthma based on the adenosine monophosphate-activated protein kinase（AMPK）/silent information regulator 1（Sirt1） pathway. [Methods] Asthmatic mice were prepared by nasal instillation of ovalbumin sensitization solution. The mice were randomly grouped into sensitization group，TPG low-dose（TPG-low，150 mg/kg TPG） group，TPG high-dose（TPG-high，300 mg/kg TPG） group，dexamethasone（4 mg/kg） group，and TPG-high+inhibitor（15 mg/kg） group. Subsequently，nebulization challenge was performed. Mice in the control group were injected with normal saline into the nasal cavity and challenged with atomized normal saline. After 1 hour，airway resistance was analyzed. Bronchoalveolar lavage fluid（BALF） was collected，and cells were classified and counted. The levels of inflammatory cytokines interleukin（IL）-6，tumor necrosis factor-α（TNF-α），and IL-17 in BALF were analyzed. The lung tissue was isolated，and the pathology of lung tissue，changes in airway mucus，AMPK phosphorylation（p-AMPK）/AMPK，and Sirt1 protein expression were detected. [Results] Compared with the control group，the expiratory pause，number of inflammatory cells such as neutrophils，the levels of IL-6，IL-17，TNF-α，inflammatory score，and PAS staining score increased in the sensitization group，while the expression of p-AMPK/AMPK and Sirt1 decreased（P<0.05）. Compared with the sensitization group，the expiratory pause，number of inflammatory cells such as neutrophils，levels of IL-6，IL-17，TNF-α，inflammatory score，and PAS staining score decreased in the TPG-low group，TPG-high group，and dexamethasone group，while the expression of p-AMPK/AMPK and Sirt1 increased（P<0.05），and significant differences were observed among TPG groups（P<0.05）. Compared with the TPG-high group，the expiratory pause，number of inflammatory cells such as neutrophils，the levels of IL-6，IL-17，TNF-α，inflammatory score，and PAS staining score increased in the TPG-high+inhibitor group，while the expression of p-AMPK/AMPK and Sirt1 decreased（P<0.05）. [Conclusion] TPG alleviates airway inflammation in mice with neutrophilic asthma by upregulating the AMPK/Sirt1 pathway.