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Adverse effects associated with expense, long-term treatment and drug resistance have made conventional therapies unfavorable, encouraging the search for alternative drugs based on plant products. In this study, the effect of <jats:italic>Calotropis procera<\/jats:italic> (Asclepiadaceae) extract against viability of promastigotes and amastigotes of <jats:italic>Leishmania major<\/jats:italic> was evaluated in vitro.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Methods<\/jats:title>\n                <jats:p>The extract from the leaves of <jats:italic>C. procera<\/jats:italic> seedlings was prepared using a methanol maceration method. The colorimetric cell viability 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to determine the growth-inhibitory effect of the extract on promastigotes. The level of reactive oxygen species (ROS) in promastigote cultures was determined after treatment with the extract using the 2',7'-dichlorofluorescein diacetate (DCFH-DA) method and compared with untreated cultures (control). After exposure to the extract the expression levels of tumor necrosis factor-\u03b1 (<jats:italic>TNF-\u03b1<\/jats:italic>), interferon gamma (<jats:italic>IFN-\u03b3<\/jats:italic>) and inducible nitric oxide synthase (<jats:italic>iNOS<\/jats:italic>) genes were determined and compared to control in peripheral blood mononuclear cells (PBMCs) infected with <jats:italic>L. major<\/jats:italic>.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Results<\/jats:title>\n                <jats:p>Based on the MTT assay, the <jats:italic>C. procera<\/jats:italic> extract significantly reduced the proliferation of <jats:italic>L. major<\/jats:italic> promastigotes with IC<jats:sub>50<\/jats:sub> values of 377.28 and 222.44\u00a0\u03bcg\/mL for 24 and 72\u00a0h, respectively (<jats:italic>p<\/jats:italic>\u2009&lt;\u20090.01). After treatment with 222.44 and 377.28\u00a0\u03bcg\/mL of <jats:italic>C. procera<\/jats:italic> extract, ROS production in <jats:italic>L. major<\/jats:italic> promastigote cultures increased 1.2- to 1.65-fold and 2- to 4-fold compared to the control, respectively (<jats:italic>p<\/jats:italic>\u2009&lt;\u20090.05). <jats:italic>C. procera<\/jats:italic> extract induced significant increases in gene expression of <jats:italic>TNF-\u03b1<\/jats:italic> (2.76\u201314.83 fold), <jats:italic>IFN-\u03b3<\/jats:italic> (25.63\u2013threefold) and <jats:italic>iNOS<\/jats:italic> (16.32\u20133.97 fold) in infected PBMCs compared to control (<jats:italic>p<\/jats:italic>\u2009&lt;\u20090.01).<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Conclusions<\/jats:title>\n                <jats:p>On the basis of its anti-leishmanial activity, <jats:italic>C. procera<\/jats:italic> can be considered as a promising new plant source for the potential treatment of leishmaniasis.<\/jats:p>\n              <\/jats:sec>","DOI":"10.1186\/s41182-024-00578-4","type":"journal-article","created":{"date-parts":[[2024,2,1]],"date-time":"2024-02-01T12:02:45Z","timestamp":1706788965000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Anti-Leishmania major activity of Calotropis procera extract by increasing ROS production and upregulating TNF-\u03b1, IFN-\u03b3 and iNOS mRNA expression under in vitro conditions"],"prefix":"10.1186","volume":"52","author":[{"given":"Shahla","family":"Amani","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Soheila","family":"Alinejad","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Negar","family":"Asadi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elham","family":"Yousefi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8753-0666","authenticated-orcid":false,"given":"Shahram","family":"Khademvatan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gordon Stanley","family":"Howarth","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2024,2,1]]},"reference":[{"key":"578_CR1","doi-asserted-by":"publisher","first-page":"164","DOI":"10.1016\/j.actatropica.2017.04.022","volume":"172","author":"M Foroutan","year":"2017","unstructured":"Foroutan M, Khademvatan S, Majidiani H, Khalkhali H, Hedayati-Rad F, Khashaveh S, Mohammadzadeh H. 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