Identification of HMOX-1-Targeting Natural Compounds in <i>Camellia nitidissima</i> Chi for NSCLC Therapy: Integrating Bioassay and In Silico Screening Approaches

<b>Background/Objectives:</b> <i>Camellia nitidissima</i> Chi (<i>C. nitidissima</i>), a traditional Chinese “food and medicine homology” plant, has demonstrated potential anti-tumor properties. However, its mechanisms of anti-lung cancer activity via ferroptosis...

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Main Authors: Lingqiu Zhang, Fan Zhang, Haimei Liang, Xiangling Qin, Chunmei Liang, Manlu Zhong, Yuemi Mo, Jinling Xie, Xiaotao Hou, Jiagang Deng, Erwei Hao, Zhengcai Du
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Language:English
Published: MDPI AG 2025-05-01
Series:Pharmaceuticals
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Online Access:https://www.mdpi.com/1424-8247/18/6/824
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author Lingqiu Zhang
Fan Zhang
Haimei Liang
Xiangling Qin
Chunmei Liang
Manlu Zhong
Yuemi Mo
Jinling Xie
Xiaotao Hou
Jiagang Deng
Erwei Hao
Zhengcai Du
author_facet Lingqiu Zhang
Fan Zhang
Haimei Liang
Xiangling Qin
Chunmei Liang
Manlu Zhong
Yuemi Mo
Jinling Xie
Xiaotao Hou
Jiagang Deng
Erwei Hao
Zhengcai Du
author_sort Lingqiu Zhang
collection DOAJ
description <b>Background/Objectives:</b> <i>Camellia nitidissima</i> Chi (<i>C. nitidissima</i>), a traditional Chinese “food and medicine homology” plant, has demonstrated potential anti-tumor properties. However, its mechanisms of anti-lung cancer activity via ferroptosis remain unclear. This study aimed to construct an integrated research system of “natural product extraction-purification, bioactivity evaluation, and computational drug screening” to explore the bioactive compounds in <i>C. nitidissima</i> leaves targeting HMOX-1-mediated ferroptosis and their anti-lung cancer mechanisms. <b>Methods:</b> Active fractions were prepared using ethanol extraction combined with polyamide column chromatography. The anti-lung cancer activity was evaluated using the NCI-H1975 cell model. Ferroptosis was verified via transmission electron microscopy (TEM), biochemical indicators, a PCR Array, and immunofluorescence. The bioactive compounds were identified using UPLC-Q Exactive MS, and their binding affinity to HMOX-1 was evaluated via molecular docking and dynamics simulations, followed by cellular validation. <b>Results:</b> The 95% F1 fraction from the extracts of <i>C. nitidissima</i> leaves exhibited the strongest anti-lung cancer activity, which could be significantly reversed by Ferrostatin-1. Furthermore, it induced typical ferroptosis-related structural damage in mitochondria, including shrinkage and a reduction in size, increased membrane density, and a reduction or even the disappearance of cristae structures. At the molecular level, this fraction significantly increased the levels of oxidative stress markers (ROS↑, MDA↑, Fe<sup>2+</sup>↑, and GSH↓) and upregulated the expression of key ferroptosis-related genes, including HMOX-1, CHAC1, and NOX1. Using UPLC-Q Exactive MS combined with computational simulation methods, four bioactive compounds with high affinity for HMOX1 were successfully identified, including isochlorogenic acid A (−8.4 kcal/mol), isochlorogenic acid C (−8.4 kcal/mol), apigenin (−7.8 kcal/mol), and chrysin (−7.3 kcal/mol). Cellular experiments validated that these compounds exhibited dose-dependent anti-proliferative effects. <b>Conclusions:</b> The leaves of <i>C. nitidissima</i> induce anti-lung cancer effects via HMOX-1-mediated ferroptosis. Isochlorogenic acid A/C, apigenin, and chrysin were identified as key bioactive components. These findings lay the foundation for the development of natural ferroptosis-targeted drugs.
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spelling doaj-art-9dc80682adc1401e9587ca5926e7514a2025-08-20T02:21:46ZengMDPI AGPharmaceuticals1424-82472025-05-0118682410.3390/ph18060824Identification of HMOX-1-Targeting Natural Compounds in <i>Camellia nitidissima</i> Chi for NSCLC Therapy: Integrating Bioassay and In Silico Screening ApproachesLingqiu Zhang0Fan Zhang1Haimei Liang2Xiangling Qin3Chunmei Liang4Manlu Zhong5Yuemi Mo6Jinling Xie7Xiaotao Hou8Jiagang Deng9Erwei Hao10Zhengcai Du11Guangxi Key Laboratory of Efficacy Study on Chinese Materia Medica, University Engineering Research Center of Reutilization of Traditional Chinese Medicine Resources, Guangxi University of Chinese Medicine, Nanning 530200, ChinaGuangxi Key Laboratory of Efficacy Study on Chinese Materia Medica, University Engineering Research Center of Reutilization of Traditional Chinese Medicine Resources, Guangxi University of Chinese Medicine, Nanning 530200, ChinaGuangxi Key Laboratory of Efficacy Study on Chinese Materia Medica, University Engineering Research Center of Reutilization of Traditional Chinese Medicine Resources, Guangxi University of Chinese Medicine, Nanning 530200, ChinaGuangxi Key Laboratory of Efficacy Study on Chinese Materia Medica, University Engineering Research Center of Reutilization of Traditional Chinese Medicine Resources, Guangxi University of Chinese Medicine, Nanning 530200, ChinaGuangxi Key Laboratory of Efficacy Study on Chinese Materia Medica, University Engineering Research Center of Reutilization of Traditional Chinese Medicine Resources, Guangxi University of Chinese Medicine, Nanning 530200, ChinaGuangxi Key Laboratory of Efficacy Study on Chinese Materia Medica, University Engineering Research Center of Reutilization of Traditional Chinese Medicine Resources, Guangxi University of Chinese Medicine, Nanning 530200, ChinaGuangxi Key Laboratory of Efficacy Study on Chinese Materia Medica, University Engineering Research Center of Reutilization of Traditional Chinese Medicine Resources, Guangxi University of Chinese Medicine, Nanning 530200, ChinaGuangxi Key Laboratory of Efficacy Study on Chinese Materia Medica, University Engineering Research Center of Reutilization of Traditional Chinese Medicine Resources, Guangxi University of Chinese Medicine, Nanning 530200, ChinaGuangxi Key Laboratory of Efficacy Study on Chinese Materia Medica, University Engineering Research Center of Reutilization of Traditional Chinese Medicine Resources, Guangxi University of Chinese Medicine, Nanning 530200, ChinaGuangxi Key Laboratory of Efficacy Study on Chinese Materia Medica, University Engineering Research Center of Reutilization of Traditional Chinese Medicine Resources, Guangxi University of Chinese Medicine, Nanning 530200, ChinaGuangxi Key Laboratory of Efficacy Study on Chinese Materia Medica, University Engineering Research Center of Reutilization of Traditional Chinese Medicine Resources, Guangxi University of Chinese Medicine, Nanning 530200, ChinaGuangxi Key Laboratory of Efficacy Study on Chinese Materia Medica, University Engineering Research Center of Reutilization of Traditional Chinese Medicine Resources, Guangxi University of Chinese Medicine, Nanning 530200, China<b>Background/Objectives:</b> <i>Camellia nitidissima</i> Chi (<i>C. nitidissima</i>), a traditional Chinese “food and medicine homology” plant, has demonstrated potential anti-tumor properties. However, its mechanisms of anti-lung cancer activity via ferroptosis remain unclear. This study aimed to construct an integrated research system of “natural product extraction-purification, bioactivity evaluation, and computational drug screening” to explore the bioactive compounds in <i>C. nitidissima</i> leaves targeting HMOX-1-mediated ferroptosis and their anti-lung cancer mechanisms. <b>Methods:</b> Active fractions were prepared using ethanol extraction combined with polyamide column chromatography. The anti-lung cancer activity was evaluated using the NCI-H1975 cell model. Ferroptosis was verified via transmission electron microscopy (TEM), biochemical indicators, a PCR Array, and immunofluorescence. The bioactive compounds were identified using UPLC-Q Exactive MS, and their binding affinity to HMOX-1 was evaluated via molecular docking and dynamics simulations, followed by cellular validation. <b>Results:</b> The 95% F1 fraction from the extracts of <i>C. nitidissima</i> leaves exhibited the strongest anti-lung cancer activity, which could be significantly reversed by Ferrostatin-1. Furthermore, it induced typical ferroptosis-related structural damage in mitochondria, including shrinkage and a reduction in size, increased membrane density, and a reduction or even the disappearance of cristae structures. At the molecular level, this fraction significantly increased the levels of oxidative stress markers (ROS↑, MDA↑, Fe<sup>2+</sup>↑, and GSH↓) and upregulated the expression of key ferroptosis-related genes, including HMOX-1, CHAC1, and NOX1. Using UPLC-Q Exactive MS combined with computational simulation methods, four bioactive compounds with high affinity for HMOX1 were successfully identified, including isochlorogenic acid A (−8.4 kcal/mol), isochlorogenic acid C (−8.4 kcal/mol), apigenin (−7.8 kcal/mol), and chrysin (−7.3 kcal/mol). Cellular experiments validated that these compounds exhibited dose-dependent anti-proliferative effects. <b>Conclusions:</b> The leaves of <i>C. nitidissima</i> induce anti-lung cancer effects via HMOX-1-mediated ferroptosis. Isochlorogenic acid A/C, apigenin, and chrysin were identified as key bioactive components. These findings lay the foundation for the development of natural ferroptosis-targeted drugs.https://www.mdpi.com/1424-8247/18/6/824<i>Camellia nitidissima</i> leavesferroptosisNSCLCbioactive screeningpreparative separation
spellingShingle Lingqiu Zhang
Fan Zhang
Haimei Liang
Xiangling Qin
Chunmei Liang
Manlu Zhong
Yuemi Mo
Jinling Xie
Xiaotao Hou
Jiagang Deng
Erwei Hao
Zhengcai Du
Identification of HMOX-1-Targeting Natural Compounds in <i>Camellia nitidissima</i> Chi for NSCLC Therapy: Integrating Bioassay and In Silico Screening Approaches
Pharmaceuticals
<i>Camellia nitidissima</i> leaves
ferroptosis
NSCLC
bioactive screening
preparative separation
title Identification of HMOX-1-Targeting Natural Compounds in <i>Camellia nitidissima</i> Chi for NSCLC Therapy: Integrating Bioassay and In Silico Screening Approaches
title_full Identification of HMOX-1-Targeting Natural Compounds in <i>Camellia nitidissima</i> Chi for NSCLC Therapy: Integrating Bioassay and In Silico Screening Approaches
title_fullStr Identification of HMOX-1-Targeting Natural Compounds in <i>Camellia nitidissima</i> Chi for NSCLC Therapy: Integrating Bioassay and In Silico Screening Approaches
title_full_unstemmed Identification of HMOX-1-Targeting Natural Compounds in <i>Camellia nitidissima</i> Chi for NSCLC Therapy: Integrating Bioassay and In Silico Screening Approaches
title_short Identification of HMOX-1-Targeting Natural Compounds in <i>Camellia nitidissima</i> Chi for NSCLC Therapy: Integrating Bioassay and In Silico Screening Approaches
title_sort identification of hmox 1 targeting natural compounds in i camellia nitidissima i chi for nsclc therapy integrating bioassay and in silico screening approaches
topic <i>Camellia nitidissima</i> leaves
ferroptosis
NSCLC
bioactive screening
preparative separation
url https://www.mdpi.com/1424-8247/18/6/824
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