{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,25]],"date-time":"2026-09-25T13:08:33Z","timestamp":1790341713548,"version":"4.1.0"},"reference-count":43,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2025,1,30]],"date-time":"2025-01-30T00:00:00Z","timestamp":1738195200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Central Council for Research in Homeopathy Research grant, Ministry of AYUSH, Govt. of India","award":["24\/2020-21\/CCRH\/Tech.\/Coll.\/DACRRI\/Haemoglobin\/3995"],"award-info":[{"award-number":["24\/2020-21\/CCRH\/Tech.\/Coll.\/DACRRI\/Haemoglobin\/3995"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Diseases"],"abstract":"<jats:p>Background\/Objectives: SARS-CoV-2 provokes acute oxidative stress in the lungs via cytokines, inflammatory mediators, and apoptotic factors, which might cause alveolar injury followed by severe respiratory syndrome during COVID-19 infection. The lack of particular antivirals for SARS-CoV-2 has opened novel avenues of complementary and alternative medicine as a potential remedy. The current study explored the mechanistic role of the ultradiluted formulation of Eupatorium (UDE) against SARS-CoV-2 recombinant S protein-mediated oxidative stress and mitochondriopathy. Methods: Cell line and BALB\/c mice were used to report that SARS-CoV-2 S protein caused an inflammatory response and subsequent cytokine storm via the NF-\u03baB pathway in the lung along with oxidative damage. Morphological examination was performed using DAPI staining and histology for treated cells and lung tissues of animals, respectively. The molecular mechanism of action of UDE was investigated through qRT-PCR for the genetic expressions of various cytokines, inflammatory, and apoptotic mediators; ELISA, immunofluorescence, immunohistochemistry, and Western blot for the translational expression of the same molecules assayed for genetic expressions; and biochemical assays for various enzymes and ROS. Results: UDE treatment suppressed the inflammatory cell infiltration and tissue-level oxidative stress and safeguarded mitochondrial integrity from free radical-mediated oxidative damage. Additionally, UDE played a direct role in restoring cellular redox homeostasis and reducing the inflammatory response by suppressing NF-\u03baB, IL-1\u03b2, IL-18, caspase-1 expression, and ROS formation. Further, a plausible mechanism of action of UDE against S protein-induced damage was proposed. Conclusions: This study described a novel therapeutic approach against S protein-mediated hyperinflammation, apoptosis, and oxidative damage. Hence, UDE may be considered as a prospective alternative to combat life-threatening consequences of SARS-CoV-2 infection.<\/jats:p>","DOI":"10.3390\/diseases13020036","type":"journal-article","created":{"date-parts":[[2025,1,30]],"date-time":"2025-01-30T04:01:39Z","timestamp":1738209699000},"page":"36","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Ultradiluted Eupatorium perfoliatum Prevents and Alleviates SARS-CoV-2 Spike Protein-Induced Lung Pathogenesis by Regulating Inflammatory Response and Apoptosis"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9134-6952","authenticated-orcid":false,"given":"Anirban","family":"Roy","sequence":"first","affiliation":[{"name":"Virology Laboratory, DAC Regional Research Institute, 50, Rajendra Chatterjee Road, Kolkata 700035, West Bengal, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Avipsha","family":"Sarkar","sequence":"additional","affiliation":[{"name":"Virology Laboratory, DAC Regional Research Institute, 50, Rajendra Chatterjee Road, Kolkata 700035, West Bengal, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Asit Kumar","family":"Roy","sequence":"additional","affiliation":[{"name":"Department of Environmental Science, University of Calcutta, 35, Ballygunge Circular Road, Kolkata 700019, West Bengal, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tanusree","family":"Ghorai","sequence":"additional","affiliation":[{"name":"Virology Laboratory, DAC Regional Research Institute, 50, Rajendra Chatterjee Road, Kolkata 700035, West Bengal, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Debadatta","family":"Nayak","sequence":"additional","affiliation":[{"name":"Central Council of Research in Homeopathy, Institutional Area, Janakpuri, New Delhi 110058, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Subhash","family":"Kaushik","sequence":"additional","affiliation":[{"name":"Central Council of Research in Homeopathy, Institutional Area, Janakpuri, New Delhi 110058, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Satadal","family":"Das","sequence":"additional","affiliation":[{"name":"Virology Laboratory, DAC Regional Research Institute, 50, Rajendra Chatterjee Road, Kolkata 700035, West Bengal, India"},{"name":"Peerless Hospital and B.K. Roy Research Centre, Kolkata 700096, West Bengal, India"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,1,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"542","DOI":"10.1016\/j.molmed.2022.04.007","article-title":"Adverse effects of COVID-19 mRNA vaccines: The spike hypothesis","volume":"28","author":"Trougakos","year":"2022","journal-title":"Trends Mol. Med."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Saini, G., and Aneja, R. (2021). Cancer as a prospective sequela of long COVID-19. Bioessays, 43.","DOI":"10.1002\/bies.202000331"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"106","DOI":"10.1016\/j.freeradbiomed.2023.05.034","article-title":"Toxicity of the spike protein of COVID-19 is a redox shift phenomenon: A novel therapeutic approach","volume":"206","author":"Schwartz","year":"2023","journal-title":"Free Rad. Biol. Med."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"271","DOI":"10.1016\/j.cell.2020.02.052","article-title":"SARS-CoV-2 Cell Entry Depends on ACE2 and TMPRSS2 and Is Blocked by a Clinically Proven Protease Inhibitor","volume":"181","author":"Hoffmann","year":"2020","journal-title":"Cell"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"e00299-20","DOI":"10.1128\/CMR.00299-20","article-title":"Critical Determinants of Cytokine Storm and Type I Interferon Response in COVID-19 Pathogenesis","volume":"34","author":"Ramasamy","year":"2021","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"e68563","DOI":"10.7554\/eLife.68563","article-title":"SARS-CoV-2 spike protein induces inflammation via TLR2-dependent activation of the NF-\u03baB pathway","volume":"10","author":"Khan","year":"2021","journal-title":"eLife"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1141","DOI":"10.1038\/s41401-020-0485-4","article-title":"Structural and functional properties of SARS-CoV-2 spike protein: Potential antivirus drug development for COVID-19","volume":"41","author":"Huang","year":"2020","journal-title":"Acta Pharmacol. Sin."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1543","DOI":"10.1134\/S0006297920120068","article-title":"COVID-19 and Oxidative Stress","volume":"85","author":"Chernyak","year":"2020","journal-title":"Biochem. Mosc."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Vollbracht, C., and Kraft, K. (2022). Oxidative Stress and Hyper-Inflammation as Major Drivers of Severe COVID-19 and Long COVID: Implications for the Benefit of High-Dose Intravenous Vitamin C. Front. Pharmacol., 13.","DOI":"10.3389\/fphar.2022.899198"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"108502","DOI":"10.1016\/j.intimp.2021.108502","article-title":"Oxidative stress and inflammatory markers in patients with COVID-19: Potential role of RAGE, HMGB1, GFAP and COX-2 in disease severity","volume":"104","author":"Passos","year":"2022","journal-title":"Int. Immunopharmacol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"191","DOI":"10.1007\/s12192-023-01330-3","article-title":"Investigation of inflammation, oxidative stress, and DNA damage in COVID-19 patients","volume":"28","author":"Temel","year":"2023","journal-title":"Cell Stress. Chaperones"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"197","DOI":"10.2217\/nnm-2022-0267","article-title":"Prophylactic and therapeutic role of catechin-loaded poly(D,L-lactic-co-glycolic acid) nanocapsules in gastric ulcer by in vitro and in vivo approach","volume":"18","author":"Roy","year":"2023","journal-title":"Nanomedicine"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1038\/s41392-022-00907-1","article-title":"The mechanism underlying extrapulmonary complications of the coronavirus disease 2019 and its therapeutic implication","volume":"7","author":"Ning","year":"2022","journal-title":"Sig. Transduct. Target. Ther."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"144","DOI":"10.1016\/j.jep.2016.05.016","article-title":"Antiviral activity of hydroalcoholic extract from Eupatorium perfoliatum L. against the attachment of influenza A virus","volume":"188","author":"Derksen","year":"2016","journal-title":"J. Ethnopharmacol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"114627","DOI":"10.1016\/j.jep.2021.114627","article-title":"In-vitro antiviral action of Eupatorium perfoliatum against dengue virus infection: Modulation of mTOR signaling and autophagy","volume":"10","author":"Sinha","year":"2022","journal-title":"J. Ethnopharmacol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"767","DOI":"10.2217\/fvl-2023-0121","article-title":"Ultradiluted Eupatorium perfoliatum Alleviates DENV-induced Fibrosis By Regulation of TGF\u03b21, MMP-9 and Interferons","volume":"18","author":"Sarkar","year":"2023","journal-title":"Future Virol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"199091","DOI":"10.1016\/j.virusres.2023.199091","article-title":"Ultradiluted SARS-CoV-2 Spike Protein mitigates hyperinflammation in lung via ferritin and MMP-9 regulation in BALB\/c mice","volume":"329","author":"Roy","year":"2023","journal-title":"Virus Res."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1007\/s12595-023-00476-3","article-title":"Safety Evaluation of Arsenicum album 6C in Cell Lines and BALB\/c Mice Through Toxicity, Proliferation and Apoptosis Studies","volume":"76","author":"Roy","year":"2023","journal-title":"Proc. Zool. Soc."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Fukushi, M., Ito, T., Oka, T., Kitazawa, T., Miyoshi-Akiyama, T., Kirikae, T., Yamashita, M., and Kudo, K. (2011). Serial histopathological examination of the lungs of mice infected with influenza A virus PR8 strain. PLoS ONE, 6.","DOI":"10.1371\/journal.pone.0021207"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1","DOI":"10.3109\/01902148.2014.956946","article-title":"Expression kinetics of metalloproteinases and their tissue inhibitors in experimental murine pulmonary tuberculosis","volume":"41","author":"Serrano","year":"2015","journal-title":"Exp. Lung Res."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1007\/978-1-61779-527-5_2","article-title":"Detection and quantification of cytokines and other biomarkers","volume":"844","author":"Chiswick","year":"2012","journal-title":"Methods Mol. Biol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1893","DOI":"10.1016\/j.freeradbiomed.2011.08.019","article-title":"Encapsulation of the flavonoid quercetin with an arsenic chelator into nanocapsules enables the simultaneous delivery of hydrophobic and hydrophilic drugs with a synergistic effect against chronic arsenic accumulation and oxidative stress","volume":"51","author":"Ghosh","year":"2011","journal-title":"Free Radic. Biol. Med."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"294","DOI":"10.1016\/j.cbi.2007.10.005","article-title":"Mitochondrial toxicity and antioxidant activity of a prenylated flavonoid isolated from Dalea elegans","volume":"171","author":"Elingold","year":"2008","journal-title":"Chem. Biol. Interact."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"331","DOI":"10.1016\/S0076-6879(84)05043-6","article-title":"Spectrophotometric detection of lipid conjugated dienes","volume":"105","author":"Glende","year":"1984","journal-title":"Methods Enzymol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1016\/S0021-9258(19)52451-6","article-title":"Protein measurement with the Folin phenol reagent","volume":"193","author":"Lowry","year":"1951","journal-title":"J. Biol. Chem."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Pulli, B., Ali, M., Forghani, R., Schob, S., Hsieh, K.L., Wojtkiewicz, G., Linnoila, J.J., and Chen, J.W. (2013). Measuring myeloperoxidase activity in biological samples. PLoS ONE, 58.","DOI":"10.1371\/journal.pone.0067976"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Klopfleisch, R. (2013). Multiparametric and semiquantitative scoring systems for the evaluation of mouse model histopathology\u2014A systematic review. BMC Vet. Res., 9.","DOI":"10.1186\/1746-6148-9-123"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1007\/978-1-0716-0211-9_16","article-title":"Histopathologic Evaluation and Scoring of Viral Lung Infection","volume":"2099","author":"Meyerholz","year":"2020","journal-title":"Methods Mol. Biol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"dmm046581","DOI":"10.1242\/dmm.046581","article-title":"Cell and animal models of SARS-CoV-2 pathogenesis and immunity","volume":"13","author":"Leist","year":"2020","journal-title":"Dis. Model. Mech."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"242","DOI":"10.1038\/s41392-022-01106-8","article-title":"Alveolar macrophages: Achilles\u2019 heel of SARS-CoV-2 infection","volume":"7","author":"Wang","year":"2022","journal-title":"Sig Transduct. Target. Ther."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"62","DOI":"10.1016\/j.cytogfr.2020.06.001","article-title":"SARS-CoV-2 infection: The role of cytokines in COVID-19 disease","volume":"54","author":"Ruiz","year":"2020","journal-title":"Cytokine Growth Factor. Rev."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"395","DOI":"10.1089\/ars.2005.7.395","article-title":"Redox regulation of NF-kappaB activation: Distinct redox regulation between the cytoplasm and the nucleus","volume":"7","author":"Kabe","year":"2005","journal-title":"Antioxid. Redox Signal"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"e2004433","DOI":"10.1002\/advs.202004433","article-title":"Cytokines: From Clinical Significance to Quantification","volume":"8","author":"Liu","year":"2021","journal-title":"Adv. Sci."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"829","DOI":"10.1038\/s41590-021-00937-x","article-title":"TLR2 senses the SARS-CoV-2 envelope protein to produce inflammatory cytokines","volume":"22","author":"Zheng","year":"2021","journal-title":"Nat. Immunol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"e21801","DOI":"10.1096\/fj.202002742RR","article-title":"Spike Protein of SARS-CoV-2 Activates Macrophages and Contributes to Induction of Acute Lung Inflammations in Mice","volume":"35","author":"Cao","year":"2021","journal-title":"FASEB J."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Hardy, E., and Fernandez-Patron, C. (2021). Targeting MMP-Regulation of Inflammation to Increase Metabolic Tolerance to COVID-19 Pathologies: A Hypothesis. Biomolecules, 11.","DOI":"10.3390\/biom11030390"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"823902","DOI":"10.1155\/2012\/823902","article-title":"Reactive oxygen species and inhibitors of inflammatory enzymes, NADPH oxidase, and iNOS in experimental models of Parkinson\u2019s disease","volume":"2012","author":"Koppula","year":"2012","journal-title":"Mediators Inflamm."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"866","DOI":"10.1038\/ni.1633","article-title":"Superoxide dismutase 1 regulates caspase-1 and endotoxic shock","volume":"9","author":"Meissner","year":"2008","journal-title":"Nat. Immunol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1352","DOI":"10.1016\/j.immuni.2019.05.020","article-title":"Caspases in Cell Death, Inflammation, and Disease","volume":"50","author":"Lamkanfi","year":"2019","journal-title":"Immunity"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"15514","DOI":"10.1073\/pnas.1414859111","article-title":"Inflammasome activation leads to Caspase-1-dependent mitochondrial damage and block of mitophagy","volume":"111","author":"Yu","year":"2014","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1007\/978-3-319-63245-2_8","article-title":"ROS Signaling in the Pathogenesis of Acute Lung Injury (ALI) and Acute Respiratory Distress Syndrome (ARDS)","volume":"967","author":"Kellner","year":"2017","journal-title":"Adv. Exp. Med. Biol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1038\/cdd.2017.186","article-title":"BCL-2 family proteins: Changing partners in the dance towards death","volume":"25","author":"Kale","year":"2018","journal-title":"Cell Death Differ."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"567","DOI":"10.1146\/annurev-immunol-073119-095439","article-title":"Caspases in Cell Death, Inflammation, and Pyroptosis","volume":"38","author":"Kesavardhana","year":"2020","journal-title":"Annul. Rev. Immunol."}],"container-title":["Diseases"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2079-9721\/13\/2\/36\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,8]],"date-time":"2025-10-08T10:38:37Z","timestamp":1759919917000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2079-9721\/13\/2\/36"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,1,30]]},"references-count":43,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2025,2]]}},"alternative-id":["diseases13020036"],"URL":"https:\/\/doi.org\/10.3390\/diseases13020036","relation":{},"ISSN":["2079-9721"],"issn-type":[{"value":"2079-9721","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,1,30]]}}}