黄芪主成分槲皮素通过MAPK通路调控PASMCs铁死亡抗低氧性肺动脉高压的研究

doi:10.5246/jcps.2024.08.053

中国药学(英文版) ›› 2024, Vol. 33 ›› Issue (8): 714-729.DOI: 10.5246/jcps.2024.08.053

• 【研究论文】 • 上一篇

黄芪主成分槲皮素通过MAPK通路调控PASMCs铁死亡抗低氧性肺动脉高压的研究

李霞¹^,²^,³, 程贝贝²^,³, 谭骏岚²^,³, 万佳婧²^,³, 王宇红⁴, 戴爱国²^,³^,^*()

^1. 湖南省中医药研究院, 湖南长沙 410208
^2. 湖南中医药大学医学院呼吸疾病研究室, 湖南长沙 410208
^3. 血管生物学与转化医学湖南省重点实验室/湖南省高校重点实验室, 湖南长沙 410208
^4. 湖南中医药大学科技创新中心, 湖南长沙 410208

收稿日期:2024-01-12 修回日期:2024-02-12 接受日期:2024-03-23 出版日期:2024-08-30 发布日期:2024-08-30
通讯作者: 戴爱国

Quercetin, the key constituent of Astragali Radix, modulates ferroptosis in PASMCs and attenuates hypoxia pulmonary hypertension via the MAPK signaling pathway

Xia Li¹^,²^,³, Beibei Cheng²^,³, Junlan Tan²^,³, Jiajing Wan²^,³, Yuhong Wang⁴, Aiguo Dai²^,³^,^*()

¹ Hunan Academy of Chinese Medicine, Changsha 410208, Hunan, China
² Department of Respiratory Diseases, Medical School, Hunan University of Chinese Medicine, Changsha 410208, Hunan, China
³ Hunan Provincial Key Laboratory of Vascular Biology and Translational Medicine, Changsha 410208, Hunan, China
⁴ Science and Technology Innovation Center, Hunan University of Chinese Medicine, Changsha 410208, Hunan, China

Received:2024-01-12 Revised:2024-02-12 Accepted:2024-03-23 Online:2024-08-30 Published:2024-08-30
Contact: Aiguo Dai
Supported by:
National Natural Science Foundation of China (Grant No. 82305214); Hunan Province’s Natural Science Fund (Grant No. 2023JJ40401); Hunan Administration of Traditional Chinese Medicine (Grant No. B2023024); Hunan Provincial Department of Education Outstanding Youth Project (Grant No. 22B0394); State Key Laboratory Project of Chinese Medicine Powder and Innovative Drugs Project (Grant No. 21PTKF1002).

摘要/Abstract

摘要：

本文基于网络药理学和实验验证探讨黄芪主成分调控铁死亡抗低氧性肺动脉高压的作用机制。利用TCMSP数据库获取黄芪活性成分及其所对应的靶标; 通过GeneCards数据库获取低氧性肺动脉的治疗靶标; 利用Venn在线工具获得黄芪活性成分和低氧性肺动脉高压的共同作用靶点。运用Cytoscape软件构建黄芪活性成分、靶标间相互作用网络关系图, 并使用CytoHubba插件获得核心靶点以及核心子网络。应用DAVID数据库进行基因本体(GO)富集分析和京都基因与基因组百科全书(KEGG)通路富集分析。检索FerrDb数据库, 获得调控铁死亡的基因并进行分析, 最后探析黄芪活性成分、低氧性肺动脉高压、铁死亡三者关系并作出预测, 并结合实验验证。结果显示, 从黄芪中筛选出18种有效活性成分, 其中槲皮素是其关键成分, 预测得到35个黄芪调控铁死亡抗低氧性肺动脉高压的可能作用靶标。进一步通过实验验证发现, 槲皮素可抑制MAPK信号通路激活, 进而抑制Fe²⁺、脂质过氧化物水平, 升高GPX4表达, 逆转铁死亡。该研究初步利用网络药理学挖掘出了黄芪调控铁死亡抗低氧性肺动脉高压的核心成分及关键信号通路; 并揭示了槲皮素作为黄芪的核心成分能够通过MAPK通路抑制PASMCs铁死亡, 逆转低氧性肺动脉高压。

关键词: 网络药理学, 黄芪, 槲皮素, PASMCs, 铁死亡, 低氧性肺动脉高压

Abstract:

This study delved into the mechanism by which the principal component of Astragali Radix regulated ferroptosis in the context of hypoxia-induced pulmonary hypertension, employing a combination of network pharmacology and experimental validation techniques. Active constituents of Astragali Radix and their corresponding targets were identified using the TCMSP database, while therapeutic targets associated with hypoxia-induced pulmonary hypertension were sourced from the GeneCards database. The Venn online tool facilitated the identification of overlapping targets between the active constituents of Astragali Radix and hypoxia-induced pulmonary hypertension. Interaction network diagrams depicting the relationship between Astragali Radix’s active constituents and their targets were constructed using Cytoscape software, with core targets and sub-networks identified using the CytoHubba plug-in. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were conducted using the DAVID database. Additionally, the FerrDb database was consulted to analyze genes implicated in regulating ferroptosis. The investigation revealed 18 active constituents selected from Astragali Radix, with quercetin emerging as the key component. A total of 35 potential targets associated with Astragali Radix in regulating ferroptosis and addressing hypoxia-induced pulmonary hypertension were predicted. Experimental validation demonstrated that quercetin could inhibit the MAPK signaling pathway, resulting in reduced Fe²⁺ and lipid peroxide levels, increased GPX4 expression, and the reversal of ferroptosis. In summary, this study elucidated the fundamental constituents and pivotal signaling pathways through which Astragali Radix modulated ferroptosis and mitigated hypoxia-induced pulmonary hypertension. Specifically, quercetin, a core constituent of Astragali Radix, was observed to inhibit ferroptosis in pulmonary arterial smooth muscle cells via the MAPK pathway and alleviate hypoxia-induced pulmonary hypertension.

Key words: Network pharmacology, Astragali Radix, Quercetin, PASMCs, Ferroptosis, Hypoxia pulmonary hypertension

Supporting:

李霞, 程贝贝, 谭骏岚, 万佳婧, 王宇红, 戴爱国. 黄芪主成分槲皮素通过MAPK通路调控PASMCs铁死亡抗低氧性肺动脉高压的研究[J]. 中国药学(英文版), 2024, 33(8): 714-729.

Xia Li, Beibei Cheng, Junlan Tan, Jiajing Wan, Yuhong Wang, Aiguo Dai. Quercetin, the key constituent of Astragali Radix, modulates ferroptosis in PASMCs and attenuates hypoxia pulmonary hypertension via the MAPK signaling pathway[J]. Journal of Chinese Pharmaceutical Sciences, 2024, 33(8): 714-729.

图/表 15

Table 1. Information on the active ingredients of Astragali Radix.

Figure 1. Common targets of Astragali Radix and anti-HPH.

Figure 2. Astragali Radix active ingredients-target network.

Table 2. Astragali Radix active ingredients-target network topology analysis results.

Figure 3. PPI network of Astragali Radix active ingredients in anti-HPH.

Figure 4. Core sub-network of Astragali Radix anti-HPH.

Table 3. MMC algorithm extracts core targets.

Figure 5. GO enrichment analysis of co-acting targets.

Figure 6. KEGG enrichment analysis of co-acting targets.

Figure 7. Target points of Astragali Radix regulating ferroptosis in the treatment of HPH.

Figure 8. Signal Pathways involved in the regulation of ferroptosis by Astragali Radix in the treatment of HPH.

Figure 9. The PPI network of Astragali Radix regulating ferroptosis in the treatment of HPH.

Figure 10. Colorimetric method to detect the effect of quercetin on intracellular Fe2+ level. Pre-incubation with quercetin (10, 30, 60, 80 and 100 μmol) were conducted lasted a period of 24/48 h, respectively. Compared with the normal group, *P < 0.05, **P < 0.01, ***P < 0.001, compared with the hypoxia group, #P < 0.05, ##P < 0.01, ###P < 0.001.

Figure 11. Effect of quercetin on lipid peroxidation in PASMCs detected by immunofluorescence. Pre-incubation with quercetin (60 μmol) was conducted and lasted a period of 48 h. Compared with the normal group, *P < 0.05, **P < 0.01, ***P < 0.001, compared with the hypoxia group, #P < 0.05, ##P < 0.01, ###P < 0.001.

Figure 12. The effects of quercetin on GPX4 and p38MAPK protein expression. Pre-incubation with quercetin (60 μmol) was conducted and lasted a period of 48 h. Compared with the normal group, *P < 0.05, **P < 0.01, ***P < 0.001, compared with the hypoxia group, #P < 0.05, ##P < 0.01, ###P < 0.001 (A: WT; B: Hypoxia; C: Quercetin; D: MAPK agonists; E: MAPK inhibitor; F: Quercetin + MAPK inhibitor).

参考文献 19

[1]	Lau, E.M.T.; Giannoulatou, E.; Celermajer, D.S.; Humbert, M. Epidemiology and treatment of pulmonary arterial hypertension. Nat. Rev. Cardiol. 2017, 14, 603–614.
[2]	Cansu D.Ü.; Korkmaz, C. Pulmonary hypertension in connective tissue diseases: epidemiology, pathogenesis, and treatment. Clin. Rheumatol. 2023, 42, 2601–2610.
[3]	Boucly, A.; Weatherald, J.; Savale, L.; Jaïs, X.; Cottin, V.; Prevot, G.; Picard, F.; de Groote, P.; Jevnikar, M.; Bergot, E.; Chaouat, A.; Chabanne, C.; Bourdin, A.; Parent, F.; Montani, D.; Simonneau, G.; Humbert, M.; Sitbon, O. Risk assessment, prognosis and guideline implementation in pulmonary arterial hypertension. Eur. Respir. J. 2017, 50, 1700889.
[4]	Xie, S.S.; Deng, Y.; Guo, S.L.; Li, J.Q.; Zhou, Y.C.; Liao, J.; Wu, D.D.; Lan, W.F. Endothelial cell ferroptosis mediates monocrotaline-induced pulmonary hypertension in rats by modulating NLRP3 inflammasome activation. Sci. Rep. 2022, 12, 3056.
[5]	Li, X.D.; Ma, M.; Deng, R.P.; Cang, X.Y.; Wu, F.Y.; Li, Y.T.; Jin, H.F. The potential effects of astragaloside on hypoxia pulmonary hypertension. Tianjin Med. J. 2021, 49, 264–268, 337.
[6]	Liu, H.; Deng, H.Y.; Tian, X.X.; Sun, H.N.; Liu, X.J.; Wang, H.X. Astragalus polysaccharides alleviates pulmonary inflammation and fibrosis in mice with hypoxia-induced pulmonary arterial hypertension by inhibiting calpain-1/ NF-κB signaling pathway. Chin. J. Pharmacol. Toxicol. 2022. 36, 98–107.
[7]	Daniel, M.C.; Menendez, C.; Moreno, E.; Moral-Sanz, J.; Barreira, B.; Galindo, P.; Pandolfi, R.; Jimenez, R.; Moreno, L.; Cogolludo, A.; Duarte, J.; Perez-Vizcaino, F. The flavonoid quercetin reverses pulmonary hypertension in rats. PLoS One. 2014, 9, e114492.
[8]	Zhang, N.; Qiu, Q.; Chen, Y.; Sun, Z.; Lu, G.; Wang, L.; Kang, P.; Wang, H. Quercetin improves pulmonary arterial hypertension in rats by regulating the HMGB1/RAGE/NF-κB pathway. J. South. Med. Univ. 2023, 43, 1606–1612.
[9]	Qin, L.Y.; Guan, P.; Wang, J.X.; Chen, Y.; Zhao, Y.S.; Yang, S.C.; Guo, Y.J.; Wang, N.; Ji, E.S. Therapeutic potential of astragaloside IV against adriamycin-induced renal damage in rats via ferroptosis. Pharmacol. 2022, 13, 812594.
[10]	Liu, Z.; Zhou, Z.; Ai, P.; Zhang, C.; Chen, J.; Wang, Y. Astragaloside IV attenuates ferroptosis after subarachnoid hemorrhage via Nrf2/HO-1 signaling pathway. Pharmacol. 2022, 13, 924826.
[11]	Liu, R.X.; Xu, C.L.; Zhang, W.L.; Cao, Y.P.; Ye, J.J.; Li, B.; Jia, S.; Weng, L.; Liu, Y.Y.; Liu, L.; Zheng, M. FUNDC1-mediated mitophagy and HIF1α activation drives pulmonary hypertension during hypoxia. Cell Death Dis. 2022, 13, 634.
[12]	Chen, T.; Sun, M.R.; Zhou, Q.; Guzman, A.M.; Ramchandran, R.; Chen, J.; Fraidenburg, D.R.; Ganesh, B.; Maienschein-Cline, M.; Obrietan, K.; Raj, J.U. MicroRNA-212-5p, an anti-proliferative miRNA, attenuates hypoxia and sugen/hypoxia-induced pulmonary hypertension in rodents. Mol. Ther. Nucleic Acids. 2022, 29, 204–216.
[13]	Zhang, F.; Liu, H.T. Identification of ferroptosis-associated genes exhibiting altered expression in pulmonary arterial hypertension. Math. Biosci. Eng. 2021, 18, 7619–7630.
[14]	Xue, T.X.; Li, L.M.; Wang, H.X. Protective effect of Astragaloside IV on hypoxia-induceed pulmonary hypertension in mice by calpain-1/NF-κB signaling pathway. Chin. J. Hosp. Pharm. 2022. 42, 1293–1298.
[15]	He, Y.Z. The therapeutic effct and mechanisms of quercetin on the hypoxia induced pulmonary hypertension. Huazhong Univ. Sci. Technol. 2016.
[16]	Cao, X.; He, Y.; Li, X.; Xu, Y.; Liu, X. The IRE1α-XBP1 pathway function in hypoxia-induced pulmonary vascular remodeling, is upregulated by quercetin, inhibits apoptosis and partially reverses the effect of quercetin in PASMCs. Am. J. Transl. Res. 2019, 11, 641–654.
[17]	He, Y.Z.; Cao, X.P.; Liu, X.S.; Li, X.C.; Xu, Y.J.; Liu, J.; Shi, J. Quercetin reverses experimental pulmonary arterial hypertension by modulating the TrkA pathway. Exp. Cell Res. 2015, 339, 122–134.
[18]	Li, J.; Cao, F.; Yin, H.L.; Huang, Z.J.; Lin, Z.T.; Mao, N.; Sun, B.; Wang, G. Ferroptosis: past, present and future. Cell Death Dis. 2020, 11, 88.
[19]	Hu, P.P.; Xu, Y.; Jiang, Y.J.; Huang, J.; Liu, Y.; Wang, D.P.; Tao, T.; Sun, Z.X.; Liu, Y. The mechanism of the imbalance between proliferation and ferroptosis in pulmonary artery smooth muscle cells based on the activation of SLC7A11. Eur. J. Pharmacol. 2022, 928, 175093.

黄芪主成分槲皮素通过MAPK通路调控PASMCs铁死亡抗低氧性肺动脉高压的研究

Quercetin, the key constituent of Astragali Radix, modulates ferroptosis in PASMCs and attenuates hypoxia pulmonary hypertension via the MAPK signaling pathway

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

图/表 15

参考文献 19

相关文章 15

编辑推荐

Metrics

本文评价

[1]	齐年景, 杨明勋, 毛姗. 基于网络药理学探讨黄芪-丹参药对治疗慢性肾小球肾炎的作用机制[J]. 中国药学(英文版), 2024, 33(7): 620-630.
[2]	魏晓玉, 于路航, 李梦茹, 徐强. 网络药理学研究揭示连花清瘟-辛夷散联合治疗新冠后嗅觉损伤的潜在作用机制[J]. 中国药学(英文版), 2024, 33(7): 631-646.
[3]	张啸, 钟叶, 胡永生, 王博龙. 《肿瘤良方大全》中肝癌用药规律挖掘及其核心药对机制分析[J]. 中国药学(英文版), 2024, 33(7): 647-658.
[4]	韩世盛, 王怡. 桃红四物汤改善动静脉内瘘失功的潜在机制: 一项网络药理学、分子对接以及分子动力学模拟研究[J]. 中国药学(英文版), 2024, 33(6): 511-524.
[5]	李双, 孙璐瑶, 尤斯涵, 张佳燚, 殷宏艳, 曹津萌, 刘心星, 郭春燕, 刘喜富. 基于网络药理学探讨桃红四物汤治疗阿尔茨海默症的主要有效成分及其作用机制[J]. 中国药学(英文版), 2024, 33(6): 525-542.
[6]	崔旭阳, 姬中杰, 时潘扬, 魏晓岑, 马玉宁, 邢梦真. 基于网络药理学探讨复方苦参注射液治疗黑色素瘤的作用机制[J]. 中国药学(英文版), 2024, 33(5): 448-457.
[7]	刘芳琳, 张春娟, 沈秋跃, 周昔程, 赵越, 富冯峰, 刘芳. 基于网络药理学和分子对接技术探讨参芪扶正注射液治疗慢性阻塞性肺疾病的潜在药理学机制[J]. 中国药学(英文版), 2024, 33(4): 339-351.
[8]	王晓航, 王乐, 肖銮娟, 芦春斌. 芒柄花黄素通过雌激素受体抑制前列腺增生[J]. 中国药学(英文版), 2024, 33(3): 216-229.
[9]	王思宇, 周淑伟, 喻斌. 基于网络药理学-分子对接探析灭幽汤"异病同治"慢性萎缩性胃炎和胃溃疡的作用机制[J]. 中国药学(英文版), 2024, 33(3): 258-271.
[10]	陈云丽, 颜仁梁, 李丽莎, 张亚敏, 徐小妹, 卢雪花, 徐榕青, 林文津. 基于网络药理学和分子对接探讨陈皮治疗阿尔茨海默病的作用机制[J]. 中国药学(英文版), 2024, 33(2): 142-155.
[11]	张弘, 张莎莎, 王焕芸, 梁越, 武世奎, 孙丽君, 夏慧敏, 白云霞, 张慧文. 基于血清药物化学和网络药理学的荜茇抗胃溃疡药效物质基础分析[J]. 中国药学(英文版), 2024, 33(2): 156-168.
[12]	王丹. 基于网络药理学探讨中药逍遥丸治疗失眠的药理机制[J]. 中国药学(英文版), 2024, 33(2): 169-177.
[13]	吴梦瑶, 刘璐, 张鹏, 张乐乐, 龚云, 杨秀伟. 基于网络药理学和实验验证研究补血益母丸治疗产后腹痛的作用机制[J]. 中国药学(英文版), 2023, 32(9): 691-703.
[14]	尚平, 刘琳, 方毅. 基于网络药理学和分子对接探讨桂枝茯苓丸治疗子宫内膜异位症的作用机制[J]. 中国药学(英文版), 2023, 32(9): 704-719.
[15]	张格第, 刘庚鑫, 晏子友. 基于meta分析和网络药理学理冲汤(丸)治疗癌症的疗效评价及作用机制研究[J]. 中国药学(英文版), 2023, 32(9): 720-735.