«上一篇/Previous Article|本期目录/Table of Contents|下一篇/Next Article»

《现代肿瘤医学》[ISSN:1672-4992/CN:61-1415/R]

期数:

2022年17期

页码:

3102-3106

栏目:

论著（基础研究）

出版日期:

2022-07-29

文章信息/Info

Title:

The effect of apple polyphenol on the LPS-induced autophagy response of lung epithelial cells by activating AMPK/SIRT1 signaling pathway

作者:

汤建华¹; 姜爱雯¹; 冯 平²; 刘克勤¹; 张志华²

1.北方学院附属第一医院药学部；2.呼吸内科，河北 张家口 075000

Author(s):

TANG Jianhua¹; JIANG Aiwen¹; FENG Ping²; LIU Keqin¹; ZHANG Zhihua²

1.Pharmaceutical Department;2.Respiratory Department,the First Affiliated Hospital of Hebei North University,Hebei Zhangjiakou 075000,China.

关键词:

苹果多酚; 脂多糖; 人肺泡上皮细胞; 腺苷酸活化蛋白激酶/沉默信息调节因子1信号通路; 自噬

Keywords:

apple polyphenol; lipopolysaccharide; human alveolar epithelial cells; AMP-activated protein kinase/Sirtuin1 signaling pathway; autophagy

分类号:

R816.4

DOI:

10.3969/j.issn.1672-4992.2022.17.007

文献标识码:

A

摘要:

目的：探讨苹果多酚通过调节腺苷酸活化蛋白激酶/沉默信息调节因子1（AMP-activated protein kinase/Sirtuin1,AMPK/SIRT1)信号通路对脂多糖（lipopolysaccharide,LPS）诱导的人肺泡上皮细胞（A549）自噬反应的影响。方法:使用不同浓度的苹果多酚提取物（apple polyphenol extract，APE）预处理A549细胞2 h后，LPS诱导A549细胞培养24 h，MTT法检测增殖活性，筛选APE最佳预处理浓度；将A549细胞分为对照组、LPS组（3 mg/L LPS）、LPS+APE组（3 mg/L LPS+20 μg/mL APE）、APE+Compound C组（3 mg/L LPS+20 μg/mL APE+50 μmol/L Compound C），免疫荧光染色观察A549细胞自噬；流式细胞术检测A549细胞凋亡；Western blot法检测细胞中自噬相关蛋白及AMPK/SIRT1通路相关蛋白表达水平。结果:与对照组比较，经LPS诱导的A549细胞增殖活性、自噬水平、LC3Ⅱ/LC3Ⅰ、Beclin-1、SIRT1、p-ULK1/ULK1、p-AMPK/AMPK蛋白表达降低，p62蛋白表达及细胞凋亡率升高（P＜0.05）；与LPS组比较，LPS+APE组细胞增殖活性、自噬水平、LC3Ⅱ/LC3Ⅰ、Beclin-1、SIRT1、p-ULK1/ULK1、p-AMPK/AMPK蛋白表达水平显著升高，p62蛋白表达及细胞凋亡率显著降低（P＜0.05）；与LPS+APE组比较，APE+Compound C组A549细胞增殖活性、自噬水平、LC3Ⅱ/LC3Ⅰ、Beclin-1、SIRT1、p-ULK1/ULK1、p-AMPK/AMPK蛋白表达水平显著降低，p62蛋白表达及细胞凋亡率显著升高（P＜0.05）。结论:苹果多酚通过激活AMPK/SIRT1 信号通路提高LPS诱导的肺上皮细胞自噬，降低细胞凋亡。

Abstract:

Objective:To investigate the effect of apple polyphenol on lipopolysaccharide（LPS)-induced autophagy in human alveolar epithelial cells（A549) by regulating AMP-activated protein kinase/Sirtuin1（AMPK/SIRT1) signaling pathway.Methods:A549 cells were pretreated with different concentrations of apple polyphenol extract（APE) for 2 h,and A549 cells were cultured for 24 h by LPS.The proliferation activity was detected by MTT method,and the optimal pretreatment concentration of APE was screened.The A549 cells were divided into control group,LPS group（3 mg/L LPS),LPS+APE group（3 mg/L LPS+20 μg/mL APE) and APE+Compound C group（3 mg/L LPS+20 μg/mL APE+50 μmol/L Compound C).Immunofluorescence staining was used to observe autophagy of A549 cells.Flow cytometry was used to detect apoptosis of A549 cells.Western blot method was used to detect the expression levels of autophagy-related proteins and AMPK/SIRT1 pathway-related proteins in cells.Results:Compared with the control group,the proliferation activity,autophagy level,LC3Ⅱ/LC3Ⅰ,Beclin-1,SIRT1,p-ULK1/ULK1,p-AMPK/AMPK protein expression of LPS-induced A549 cells decreased,and the p62 protein expression and the apoptosis rate increased（P＜0.05).Compared with the LPS group,the proliferation activity,autophagy level,LC3Ⅱ/LC3Ⅰ,Beclin-1,SIRT1,p-ULK1/ULK1,p-AMPK/AMPK protein expression in the LPS+APE group increased,and the p62 protein expression and the apoptosis rate decreased（P＜0.05).Compared with the LPS+APE group,the proliferation activity,autophagy level,LC3Ⅱ/LC3Ⅰ,Beclin-1,SIRT1,p-ULK1/ULK1,p-AMPK/AMPK protein expression in the APE+Compound C group decreased,and the p62 protein expression and the apoptosis rate increased（P＜0.05).Conclusion:Apple polyphenol can increase LPS-induced autophagy of lung epithelial cells and reduce cell apoptosis by activating AMPK/SIRT1 signaling pathway.

参考文献/References

［1］ GROSS CM,KKELLNER M,WANG T,et al.LPS-induced acute lung injury involves NF-κB-mediated downregulation of SOX18［J］.Am J Respir Cell Mol Biol,2018,58(5):614-624.
［2］ 古丽尼歌尔?阿布都米吉提,沙依拜?沙比提,丛媛媛,等.基于Nrf2通路探讨阿里红三萜酸减轻脂多糖诱导的小鼠急性肺损伤的作用［J］.中国病理生理杂志,2021,37(1):112-118. ABUDUMIJITI G,SHABITI S,CONG YY.Triterpenic acid of fomes officinalis ames attenuates LPS-induced acute lung injury in mice via Nrf2 pathway［J］.Chinese Journal of Pathophysiology,2021,37(1):112-118.
［3］ 秦莉,张红.细胞自噬在急性肺损伤中作用机制的研究进展［J］.山东医药,2021,61(17):98-100. QIN L,ZHANG H.Research progress on the mechanism of autophagy in acute lung injury［J］.Shandong Medical Journal,2021,61(17):98-100.
［4］ 章书豪,邵思铭,陈方政,等.苹果多酚对野百合碱诱导的肺动脉高压大鼠肺血管重构的作用和机制［J］.中国应用生理学杂志,2019,35(3):209-214. ZHANG SH,SHAO SM,CHEN FZ,et al.Effects of apple polyphenols on monocrotaline-induced pulmonary vascular remodeling in rats and its mechanism［J］.Chinese Journal of Applied Physiology,2019,35(3):209-214.
［5］ SHARMA S,RANA S,PATIAL V,et al.Antioxidant and hepatoprotective effect of polyphenols from apple pomace extract via apoptosis inhibition and Nrf2 activation in mice［J］.Hum Exp Toxicol,2016,35(12):1264-1275.
［6］ HUANG XT,LIU W,ZHOU Y,et al.Galectin-1 ameliorates lipopolysaccharide-induced acute lung injury via AMPK-Nrf2 pathway in mice［J］.Free Radic Biol Med,2020,146(1):222-233.
［7］ HUA C,ZHAO J,WANG H,et al.Apple polyphenol relieves hypoxia-induced pulmonary arterial hypertension via pulmonary endothelium protection and smooth muscle relaxation:In vivo and in vitro studies［J］.Biomed Pharmacother,2018,107(1):937-944.
［8］ WU KH,HO CT,CHEN ZF,et al.The apple polyphenol phloretin inhibits breast cancer cell migration and proliferation via inhibition of signals by type 2 glucose transporter［J］.J Food Drug Anal,2018,26(1):221-231.
［9］ 史新阳,高晓慧,赵蓓,等.W3D通过调控TLR4/MD2-NF-κB p65信号通路改善LPS诱导的小鼠急性肺损伤［J］.中国药理学通报,2021,37(5):657-661. SHI XY,GAO XH,ZHAO B,et al.W3D improves LPS-induced acute lung injury in mice by regulating TLR4/MD2-NF-κB p65 signaling pathway［J］.Chinese Pharmacological Bulletin,2021,37(5):657-661.
［10］ LI D,CUI Y,WANG X,et al.Apple polyphenol extract alleviates lipid accumulation in free-fatty-acid-exposed HepG2 cells via activating autophagy mediated by SIRT1/AMPK signaling［J］.Phytother Res,2021,35(3):1416-1431.
［11］ 梁慧,杨慧敏,王潇.青天葵黄酮F通过促进自噬减轻脂多糖诱导的大鼠急性肺损伤［J］.广西医科大学学报,2021,38(5):837-846. LIANG H,YANG HM,WANG X.Nervilifordin F alleviates lipopolysaccharide-induced acute lung injury in rats via promotion of autophagy［J］.Journal of Guangxi Medical University,2021,38(5):837-846.
［12］ ZHANG X,ZHENG J,YAN Y,et al.Angiotensin-converting enzyme 2 regulates autophagy in acute lung injury through AMPK/mTOR signaling［J］.Arch Biochem Biophys,2019,672(1):108061.
［13］ FU C,HAO S,XU X,et al.Activation of SIRT1 ameliorates LPS-induced lung injury in mice via decreasing endothelial tight junction permeability［J］.Acta Pharmacol Sin,2019,40(5):630-641.
［14］ LI S,QIAN Q,YING N,et al.Activation of the AMPK-SIRT1 pathway contributes to protective effects of Salvianolic acid A against lipotoxicity in hepatocytes and NAFLD in mice［J］.Front Pharmacol,2020,11(1):1-15.
［15］ LI X,JAMAL M,GUO P,et al.Irisin alleviates pulmonary epithelial barrier dysfunction in sepsis-induced acute lung injury via activation of AMPK/SIRT1 pathways［J］.Biomed Pharmacother,2019,18(1):1-10.

备注/Memo

备注/Memo:

河北省政府资助临床医学优秀人才培养项目计划(编号：39003-2)

常用功能

更新日期/Last Update: 2022-07-29