{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T20:33:52Z","timestamp":1791405232086,"version":"4.3.3"},"reference-count":42,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2024,5,11]],"date-time":"2024-05-11T00:00:00Z","timestamp":1715385600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Science and Qinghai Province Science and Technology Department project","award":["2023-ZJ-992Q"],"award-info":[{"award-number":["2023-ZJ-992Q"]}]},{"name":"Qinghai Kunlun Talents\u2014High-end Innovation and Entrepreneurship Talents Project","award":["2023-ZJ-992Q"],"award-info":[{"award-number":["2023-ZJ-992Q"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["TropicalMed"],"abstract":"<jats:p>Alveolar echinococcosis (AE) stands as a perilous zoonotic affliction caused by the larvae of Echinococcus multilocularis. There is an imperative need to explore novel therapeutic agents or lead compounds for the treatment of AE. Asparagusic acid, characterized by its low toxicity and possessing antimicrobial, antioxidant, and anti-parasitic attributes, emerges as a promising candidate. The aim of this study was to investigate the in vivo and in vitro efficacy of asparagusic acid against E. multilocularis. Morphological observations, scanning electron microscopy, ROS assays, mitochondrial membrane potential assays, and Western blot were used to evaluate the in vitro effects of asparagusic acid on protoscoleces. The effects of asparagusic acid on vesicles were assessed via PGI release, \u03b3-GGT release, and transmission electron microscopy observations. CellTiter-Glo assays, Caspase3 activity assays, flow cytometry, and Western blot were used for an evaluation of the effect of asparaginic acid on the proliferation and apoptosis of germinal cells. The in vivo efficacy of asparagusic acid was evaluated in a murine AE model. Asparagusic acid exhibited a pronounced killing effect on the protoscoleces post-treatment. Following an intervention with asparagusic acid, there was an increase in ROS levels and a decline in mitochondrial membrane potential in the protoscolex. Moreover, asparagusic acid treatment resulted in the upregulation of PGI and \u03b3-GGT release in metacestode vesicles, concomitant with the inhibition of germinal cell viability. Furthermore, asparagusic acid led to an enhanced relative expression of Caspase3 in the culture supernatant of both the protoscoleces and germinal cells, accompanied by an increase in the proportion of apoptotic germinal cells. Notably, asparagusic acid induced an augmentation in Bax and Caspase3 protein expression while reducing Bcl2 protein expression in both the protoscoleces and germinal cells. In vitro cytotoxicity assessments demonstrated the low toxicity of asparagusic acid towards normal human hepatocytes and HFF cells. Additionally, in vivo experiments revealed that asparagusic acid administration at doses of 10 mg\/kg and 40 mg\/kg significantly reduced metacestode wet weight. A histopathological analysis displayed the disruption of the germinal layer structure within lesions post-asparagusic acid treatment, alongside the preservation of laminated layer structures. Transmission electron microscopy further revealed mitochondrial swelling and heightened cell necrosis subsequent to the asparagusic acid treatment. Furthermore, asparagusic acid promoted Caspase3 and Bax protein expression while decreasing Bcl2 protein expression in perilesional tissues. Subsequently, it inhibited the expression of Ki67, MMP2, and MMP9 proteins in the perilesional tissues and curbed the activation of the PI3K\/Akt signaling pathway within the lesion-host microenvironmental tissues. Asparagusic acid demonstrated a pronounced killing effect on E. multilocularis, suggesting its potential as a promising therapeutic agent for the management of AE.<\/jats:p>","DOI":"10.3390\/tropicalmed9050110","type":"journal-article","created":{"date-parts":[[2024,5,13]],"date-time":"2024-05-13T03:34:01Z","timestamp":1715571241000},"page":"110","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Efficacy and Safety of Asparagusic Acid against Echinococcus multilocularis In Vitro and in a Murine Infection Model"],"prefix":"10.3390","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1148-0452","authenticated-orcid":false,"given":"Zhuanhong","family":"Lu","sequence":"first","affiliation":[{"name":"School of Clinical Medicine, Qinghai University Affiliated Hospital, Xining 810001, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yating","family":"Wang","sequence":"additional","affiliation":[{"name":"School of Clinical Medicine, Qinghai University Affiliated Hospital, Xining 810001, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chuanchuan","family":"Liu","sequence":"additional","affiliation":[{"name":"School of Clinical Medicine, Qinghai University Affiliated Hospital, Xining 810001, China"},{"name":"Qinghai Key Laboratory of Echinococcosis Research, Xining 810001, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haining","family":"Fan","sequence":"additional","affiliation":[{"name":"School of Clinical Medicine, Qinghai University Affiliated Hospital, Xining 810001, China"},{"name":"Qinghai Key Laboratory of Echinococcosis Research, Xining 810001, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,5,11]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"e12711","DOI":"10.1111\/pim.12711","article-title":"Echinococcus multilocularis vesicular fluid induces the expression of immune checkpoint proteins in vitro","volume":"42","author":"Bellanger","year":"2020","journal-title":"Parasite Immunol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"315","DOI":"10.1016\/bs.apar.2016.11.001","article-title":"Global distribution of alveolar and cystic echinococcosis","volume":"95","author":"Deplazes","year":"2017","journal-title":"Adv. Parasitol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"e00075-18","DOI":"10.1128\/CMR.00075-18","article-title":"Echinococcosis: Advances in the 21st century","volume":"32","author":"Wen","year":"2019","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"2529","DOI":"10.1007\/s00268-015-3109-2","article-title":"Outcomes after liver resection for hepatic alveolar echinococcosis: A single-center cohort study","volume":"39","author":"Joliat","year":"2015","journal-title":"World J. Surg."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"72","DOI":"10.1016\/j.jhep.2008.03.023","article-title":"Alveolar echinococcosis: From a deadly disease to a well-controlled infection. Relative survival and economic analysis in Switzerland over the last 35 years","volume":"49","author":"Torgerson","year":"2008","journal-title":"J. Hepatol."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Xu, X., Qian, X., Gao, C., Pang, Y., Zhou, H., Zhu, L., Wang, Z., Pang, M., Wu, D., and Yu, W. (2022). Advances in the pharmacological treatment of hepatic alveolar echinococcosis: From laboratory to clinic. Front. Microbiol., 13.","DOI":"10.3389\/fmicb.2022.953846"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"2995","DOI":"10.1007\/s00436-016-5054-x","article-title":"Intra-cystic concentrations of albendazole-sulphoxide in human cystic echinococcosis: A systematic review and analysis of individual patient data","volume":"115","author":"Naderer","year":"2016","journal-title":"Parasitol. Res."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"70","DOI":"10.1051\/parasite\/2014073","article-title":"Treatment of echinococcosis: Albendazole and mebendazole--what else?","volume":"21","author":"Hemphill","year":"2014","journal-title":"Parasite"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"3069","DOI":"10.2147\/IJN.S397526","article-title":"Oral delivery of anti-parasitic agent-loaded plga nanoparticles: Enhanced liver targeting and improved therapeutic effect on hepatic alveolar echinococcosis","volume":"18","author":"Li","year":"2023","journal-title":"Int. J. Nanomed."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"129903","DOI":"10.1016\/j.foodchem.2021.129903","article-title":"Improving the bioactive ingredients and functions of asparagus from efficient to emerging processing technologies: A review","volume":"358","author":"Yu","year":"2021","journal-title":"Food Chem."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Jaramillo-Carmona, S., Guill\u00e9n-Bejarano, R., Jim\u00e9nez-Araujo, A., Rodr\u00edguez-Arcos, R., and L\u00f3pez, S. (2018). In vitro toxicity of asparagus saponins in distinct multidrug-resistant colon cancer cells. Chem. Biodivers., 15.","DOI":"10.1002\/cbdv.201800282"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1016\/j.phytochem.2013.09.014","article-title":"Asparagusic acid","volume":"97","author":"Mitchell","year":"2014","journal-title":"Phytochemistry"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"394","DOI":"10.5059\/yukigoseikyokaishi.35.394","article-title":"Malformin and Asparagusic Acids as Sulfur-containing Compounds Having Biological Activities to Plants","volume":"35","author":"Iriuchijima","year":"1977","journal-title":"J. Synth. Org. Chem. Jpn."},{"key":"ref_14","first-page":"43","article-title":"Identification of asparagusic acid as a nematicide occuring naturally in the roots of asparagus","volume":"6","author":"Takasugi","year":"1975","journal-title":"Chem. Informationsdienst"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"450","DOI":"10.1186\/s13071-021-04952-7","article-title":"In vitro and in vivo efficacy of thiacloprid against Echinococcus multilocularis","volume":"14","author":"Liu","year":"2021","journal-title":"Parasit. Vectors"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1016\/j.biopha.2018.06.080","article-title":"Valproic acid attenuates A\u03b225-35-induced neurotoxicity in PC12 cells through suppression of mitochondria-mediated apoptotic pathway","volume":"106","author":"Zhao","year":"2018","journal-title":"Biomed. Pharmacother."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2779","DOI":"10.1080\/21655979.2021.1930335","article-title":"Trichosanthin inhibits cervical cancer by regulating oxidative stress-induced apoptosis","volume":"12","author":"Zhu","year":"2021","journal-title":"Bioengineered."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1016\/j.canlet.2018.05.046","article-title":"Targeting autophagy enhances apatinib-induced apoptosis via endoplasmic reticulum stress for human colorectal cancer","volume":"431","author":"Cheng","year":"2018","journal-title":"Cancer Lett."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"919","DOI":"10.2478\/s11686-020-00231-0","article-title":"Prediction and Identification of Epitopes in the Emy162 Antigen of Echinococcus multilocularis","volume":"65","author":"Pang","year":"2020","journal-title":"Acta Parasitol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"114","DOI":"10.1016\/j.ijpddr.2023.03.002","article-title":"Investigation of the mechanism of action of mefloquine and derivatives against the parasite Echinococcus multilocularis","volume":"21","author":"Memedovski","year":"2023","journal-title":"Int. J. Parasitol.-Drugs"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"451","DOI":"10.1093\/jac\/46.3.451","article-title":"Benzimidazoles in the treatment of alveolar echinococcosis: A comparative study and review of the literature","volume":"46","author":"Reuter","year":"2000","journal-title":"J. Antimicrob. Chemother."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"585","DOI":"10.1093\/jpp\/rgac103","article-title":"Research progress on pharmacological effects of isoalantolactone","volume":"75","author":"Xu","year":"2023","journal-title":"J. Pharm. Pharmacol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"629","DOI":"10.1021\/acs.jnatprod.5b01055","article-title":"Natural products as sources of new drugs from 1981 to 2014","volume":"79","author":"Newman","year":"2016","journal-title":"J. Nat. Prod."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"204","DOI":"10.1016\/j.drudis.2015.01.009","article-title":"An analysis of FDA-approved drugs: Natural products and their derivatives","volume":"21","author":"Patridge","year":"2016","journal-title":"Drug Discov. Today"},{"key":"ref_25","first-page":"398","article-title":"Traditional Chinese medicine for treatment of echinococcosis: A review","volume":"35","author":"Chen","year":"2023","journal-title":"Zhongguo Xue Xi Chong Bing Fang Zhi Za Zhi"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"2977","DOI":"10.1016\/j.bbamcr.2016.09.012","article-title":"Activation of apoptosis signalling pathways by reactive oxygen species","volume":"1863","year":"2016","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1394","DOI":"10.1080\/01635581.2016.1224362","article-title":"Seed oil of brucea javanicainduces cell cycle arrest and apoptosis via reactive oxygen species-mediated mitochondrial dysfunction in human lung cancer cells","volume":"68","author":"Wang","year":"2016","journal-title":"Nutr. Cancer"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"16096","DOI":"10.1038\/cddiscovery.2016.96","article-title":"Matrine induces RIP3-dependent necroptosis in cholangiocarcinoma cells","volume":"3","author":"Xu","year":"2017","journal-title":"Cell Death Discov."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Al-Ishaq, R.K., Overy, A.J., and B\u00fcsselberg, D. (2020). Phytochemicals and gastrointestinal cancer: Cellular mechanisms and effects to change cancer progression. Biomolecules, 10.","DOI":"10.3390\/biom10010105"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"108280","DOI":"10.1016\/j.exppara.2022.108280","article-title":"In vitro protoscolicidal effects of lithocholic acid on protoscoleces of Echinococcus granulosus and its mechanism","volume":"239","author":"Xu","year":"2022","journal-title":"Exp. Parasitol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"269","DOI":"10.1007\/s12291-013-0408-y","article-title":"Alkaline phosphatase: An overview","volume":"29","author":"Sharma","year":"2014","journal-title":"Indian J. Clin. Biochem."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1016\/j.ijpddr.2020.06.002","article-title":"Drug repurposing applied: Activity of the anti-malarial mefloquine against Echinococcus multilocularis","volume":"13","author":"Rufener","year":"2020","journal-title":"Int. J. Parasitol.-Drug"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/bs.apar.2016.09.005","article-title":"Immunology of Alveolar and Cystic Echinococcosis (AE and CE)","volume":"96","author":"Gottstein","year":"2017","journal-title":"Adv. Parasitol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"72","DOI":"10.1051\/parasite\/2014070","article-title":"On the importance of targeting parasite stem cells in anti-echinococcosis drug development","volume":"21","author":"Brehm","year":"2014","journal-title":"Parasite"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1016\/j.actatropica.2014.08.007","article-title":"Efficacy of albendazole in combination with thymol against Echinococcus multilocularis protoscoleces and metacestodes","volume":"140","author":"Celina","year":"2014","journal-title":"Acta Trop."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Wang, W., Zhu, M., Xu, Z., Li, W., Dong, X., Chen, Y., Lin, B., and Li, M. (2019). Ropivacaine promotes apoptosis of hepatocellular carcinoma cells through damaging mitochondria and activating caspase-3 activity. Biol. Res., 52.","DOI":"10.1186\/s40659-019-0242-7"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"2280","DOI":"10.1021\/acschembio.1c00456","article-title":"Deorphanizing Caspase-3 and Caspase-9 substrates in and out of apoptosis with deep substrate profiling","volume":"16","author":"Araya","year":"2021","journal-title":"ACS Chem. Biol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"553","DOI":"10.1159\/000480528","article-title":"Quercetin inhibits cell migration and invasion in human osteosarcoma cells","volume":"43","author":"Lan","year":"2017","journal-title":"Cell. Physiol. Biochem."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/bs.pmbts.2017.02.005","article-title":"Biochemical and biological attributes of matrix metalloproteinases","volume":"147","author":"Cui","year":"2017","journal-title":"Prog. Mol. Biol. Transl. Sci."},{"key":"ref_40","first-page":"e919757","article-title":"S100A16 Regulates HeLa cell through the phosphatidylinositol 3 kinase (PI3K)\/AKT signaling pathway","volume":"26","author":"Zhang","year":"2020","journal-title":"Med. Sci. Monit."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"409","DOI":"10.1080\/20477724.2022.2104055","article-title":"Echinococcus multilocularis protoscoleces enhance glycolysis to promote M2 macrophages through PI3K\/Akt\/mTOR signaling pathway","volume":"117","author":"Zhang","year":"2023","journal-title":"Pathog. Glob. Health"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"106084","DOI":"10.1016\/j.actatropica.2021.106084","article-title":"Microcyst fluid promotes the migration and invasion of fibroblasts in the adventitial layer of alveolar echinococcosis","volume":"223","author":"Liu","year":"2021","journal-title":"Acta Trop."}],"container-title":["Tropical Medicine and Infectious Disease"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2414-6366\/9\/5\/110\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T14:40:54Z","timestamp":1760107254000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2414-6366\/9\/5\/110"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,5,11]]},"references-count":42,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2024,5]]}},"alternative-id":["tropicalmed9050110"],"URL":"https:\/\/doi.org\/10.3390\/tropicalmed9050110","relation":{},"ISSN":["2414-6366"],"issn-type":[{"value":"2414-6366","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,5,11]]}}}