杨梅素通过激活Sirt1调节JNK/Smad3通路改善哮喘气道炎症和重塑
2024/10/29
摘要
背景：杨梅素具有多种生物活性和健康益处；然而，其对哮喘气道重塑的影响尚未见报道。本研究旨在探讨杨梅素通过激活Sirt1改善气道重塑的可能性，并有可能成为哮喘的新治疗方法。
方法:用脂多糖刺激RAW 264.7细胞并与3T6细胞体外共培养，模拟体内炎症对气道重塑的影响。使用卵清蛋白诱导的慢性哮喘小鼠模型，我们使用蛋白质印迹法和定量PCR比较杨梅素和/或Sirt1抑制剂EX-527治疗下炎症因子和气道重塑相关因子的变化。将携带Smad3位点突变的表达质粒转染3T6细胞，鉴定Smad3蛋白上Sirt1的去甲基化位点。
结果:杨梅素显著减少哮喘小鼠气道炎症细胞的浸润和白细胞介素(IL)-6、IL-5的产生，抑制杯状细胞黏液分泌、胶原纤维增殖和支气管肺泡灌洗液中炎症细胞的增加。体外实验结果与体内实验一致。探究杨梅素的作用机制，我们发现杨梅素通过激活Sirt1来下调体内和体外磷酸化(p)-JNK， p-Smad3和乙酰化Smad3蛋白的水平。杨梅素激活的Sirt1鉴定出K341是Smad3的主要去乙酰化位点。
结论:杨梅素通过激活Sirt1调节JNK/Smad3通路改善哮喘气道炎症和重塑。
Myricetin ameliorates airway inflammation and remodeling in asthma by activating Sirt1 to regulate the JNK/Smad3 pathway
D. Huang, S. Y. Bai, G. Q. Qiu, C. Jiang, M. Huang, Y. Wang, et al.
Abstract
BACKGRUND:Myricetin has various biological activities and health benefits; however, its effects on airway remodeling in asthma have not been reported. We aimed to investigate the possibility that myricetin improves airway remodeling by activating Sirt1 and has potential as a new treatment for asthma.
METHODS: RAW 264.7 cells were stimulated with lipopolysaccharide and co-cultured with 3T6 cells in vitro to simulate the in vivo effects of inflammation on airway remodeling. Using an ovalbumin-induced chronic asthma mouse model, we compared changes in inflammatory factors and airway remodeling-related factors under treatment with myricetin and/or the Sirt1 inhibitor EX-527 using western blotting and quantitative PCR. Expression plasmids carrying Smad3 site mutations were transfected into 3T6 cells to identify the Sirt1 deace tylation site on Smad3 protein.
RESULTS: Myricetin significantly reduced the infiltration of airway inflammatory cells and the production of interleukin (IL)-6 and IL-5, and inhibited mucus secretion by goblet cells, collagen fiber proliferation, and the increase in inflammatory cells in bronchoalveolar lavage fluid from asthmatic mice. Results of in vitro experi ments were consistent with those conducted in vivo. Exploring the mechanism of action of myricetin, we found that myricetin downregulated the levels of phosphorylated (p)-JNK, p-Smad3, and acetylated Smad3 proteins by activating Sirt1 both in vivo and in vitro. K341 was identified as the main deacetylation site of Smad3 by myricetin-activated Sirt1.
CONCLUSIONS: Myricetin ameliorates airway inflammation and remodeling in asthma by activating Sirt1 to regulate the JNK/Smad3 pathway.
背景：杨梅素具有多种生物活性和健康益处；然而，其对哮喘气道重塑的影响尚未见报道。本研究旨在探讨杨梅素通过激活Sirt1改善气道重塑的可能性，并有可能成为哮喘的新治疗方法。
方法:用脂多糖刺激RAW 264.7细胞并与3T6细胞体外共培养，模拟体内炎症对气道重塑的影响。使用卵清蛋白诱导的慢性哮喘小鼠模型，我们使用蛋白质印迹法和定量PCR比较杨梅素和/或Sirt1抑制剂EX-527治疗下炎症因子和气道重塑相关因子的变化。将携带Smad3位点突变的表达质粒转染3T6细胞，鉴定Smad3蛋白上Sirt1的去甲基化位点。
结果:杨梅素显著减少哮喘小鼠气道炎症细胞的浸润和白细胞介素(IL)-6、IL-5的产生，抑制杯状细胞黏液分泌、胶原纤维增殖和支气管肺泡灌洗液中炎症细胞的增加。体外实验结果与体内实验一致。探究杨梅素的作用机制，我们发现杨梅素通过激活Sirt1来下调体内和体外磷酸化(p)-JNK， p-Smad3和乙酰化Smad3蛋白的水平。杨梅素激活的Sirt1鉴定出K341是Smad3的主要去乙酰化位点。
结论:杨梅素通过激活Sirt1调节JNK/Smad3通路改善哮喘气道炎症和重塑。
（中日友好医院呼吸与危重症医学科 李春晓 摘译 林江涛 审校）
（Phytomedicine 2024 Vol. 135 DOI: ARTN 15604410.1016/j.phymed.2024.156044）
（Phytomedicine 2024 Vol. 135 DOI: ARTN 15604410.1016/j.phymed.2024.156044）
Myricetin ameliorates airway inflammation and remodeling in asthma by activating Sirt1 to regulate the JNK/Smad3 pathway
D. Huang, S. Y. Bai, G. Q. Qiu, C. Jiang, M. Huang, Y. Wang, et al.
Abstract
BACKGRUND:Myricetin has various biological activities and health benefits; however, its effects on airway remodeling in asthma have not been reported. We aimed to investigate the possibility that myricetin improves airway remodeling by activating Sirt1 and has potential as a new treatment for asthma.
METHODS: RAW 264.7 cells were stimulated with lipopolysaccharide and co-cultured with 3T6 cells in vitro to simulate the in vivo effects of inflammation on airway remodeling. Using an ovalbumin-induced chronic asthma mouse model, we compared changes in inflammatory factors and airway remodeling-related factors under treatment with myricetin and/or the Sirt1 inhibitor EX-527 using western blotting and quantitative PCR. Expression plasmids carrying Smad3 site mutations were transfected into 3T6 cells to identify the Sirt1 deace tylation site on Smad3 protein.
RESULTS: Myricetin significantly reduced the infiltration of airway inflammatory cells and the production of interleukin (IL)-6 and IL-5, and inhibited mucus secretion by goblet cells, collagen fiber proliferation, and the increase in inflammatory cells in bronchoalveolar lavage fluid from asthmatic mice. Results of in vitro experi ments were consistent with those conducted in vivo. Exploring the mechanism of action of myricetin, we found that myricetin downregulated the levels of phosphorylated (p)-JNK, p-Smad3, and acetylated Smad3 proteins by activating Sirt1 both in vivo and in vitro. K341 was identified as the main deacetylation site of Smad3 by myricetin-activated Sirt1.
CONCLUSIONS: Myricetin ameliorates airway inflammation and remodeling in asthma by activating Sirt1 to regulate the JNK/Smad3 pathway.
