Ethanolic seed extract of Artocarpus heterophyllus ameliorates histomorphological, morphometric and functional alterations in the rat testis induced by sildenafil citrate 

Ethanolic seed extract of Artocarpus heterophyllus ameliorates histomorphological, morphometric and functional alterations in the rat testis induced by sildenafil citrate | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Ethanolic seed extract of Artocarpus heterophyllus ameliorates histomorphological, morphometric and functional alterations in the rat testis induced by sildenafil citrate John Juma Ochieng, Jackline Namulema, Onanuga Ismail Olasile, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6413247/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background: One in every ten men is affected by ED regardless of their age. Sildenafil citrate is the first-line drug in the treatment of ED and has been associated with seminiferous tubules and testicular degeneration, translating into decreased spermatogenesis. The study evaluated the effect of ethanolic seed extract of Artocarpus heterophyllus on changes in testicular histology, seminal fluid and serum testosterone levels alone and in ameliorating testicular changes induced by Sildenafil Citrate. Methods: Thirty-six adult male Wistar rats were randomly divided into six groups (n=6) and orally dosed daily with NS, seed extract of AH (500 or 1000 mg/kg), SC (50 mg/kg) or AH+SC (1000 mg/kg + 50 mg/kg or 1000 mg/kg + 100 mg/kg). Testicular histopathological and semen analyses were performed via H&E, PAS and Masson’s trichome staining and microscopy. Differences in sperm parameters, testicular size, seminiferous tubule characteristics, Leydig cell volume, and serum testosterone levels were determined. Results: SC administration significantly decreased the sperm count (3.47±1.46), motility (23.414±6.63) and morphology (0.038±0.02) compared to the negative control (6.04±0.47, 27.888±0.81 and 0.058±0.02, respectively). The administration of AH 1000 to SC-positive rats improved these parameters in an SC dose-dependent manner, i.e., 10.54±1.48, 31.516±0.99 and 0.0781±0.02 and 9.36±0.36, 27.314±1.45 and 0.049±0.03 for 50 mg/kg and 100 mg/kg SC, respectively. AH improved the diameter of the seminiferous tubules but decreased the height in SC-positive rats [(297.2±21.64) and (289.0±8.60)] and [100.2±5.03 and 96.8±4.20] compared with SC alone (194.0±51.05 and 60.0±15.26, respectively). SC decreased the Leydig cell diameter (6696.05±2525.53), AH 1000 significantly reversed this (8218.53±1291.49) to above those of the negative control (7584.18±816.46). Serum testosterone levels were lower in the SC group (2.53±1.88) than in the normal group (3.28±1.74), but these levels were improved by AH (4.596±0.73 and 5.46±2.55). Histological examination of the testes revealed increased spermatogenesis and Leydig cell proliferation with a higher dose of extract. Conclusion: This study revealed that the ethanolic seed extract of A. heterophyllus affects male rat reproductive system function by improving sperm parameters, testicular volume, seminiferous tubule characteristics, Leydig cell volume and serum testosterone levels. AH also improved all these parameters in SC-positive rats in an SC dose-dependent manner. ackfruit Artocarpus heterophyllus sildenafil citrate male fertility testicular morphology serum testosterone Leydig cells seminiferous tubules Figures Figure 1 Figure 2 Figure 3 Full Text Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6413247","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":477898537,"identity":"8f73ee16-7788-49ff-afdf-8d965970329b","order_by":0,"name":"John Juma Ochieng","email":"","orcid":"","institution":"Kampala International University","correspondingAuthor":false,"prefix":"","firstName":"John","middleName":"Juma","lastName":"Ochieng","suffix":""},{"id":477898538,"identity":"b5d40e87-71bf-452a-bf86-608f5fc619d5","order_by":1,"name":"Jackline Namulema","email":"data:image/png;base64,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","orcid":"","institution":"Uzima University – Kisumu","correspondingAuthor":true,"prefix":"","firstName":"Jackline","middleName":"","lastName":"Namulema","suffix":""},{"id":477898539,"identity":"44f334c7-409b-4c58-ba0e-28da76469b8e","order_by":2,"name":"Onanuga Ismail Olasile","email":"","orcid":"","institution":"Olabisi Onabanjo University","correspondingAuthor":false,"prefix":"","firstName":"Onanuga","middleName":"Ismail","lastName":"Olasile","suffix":""},{"id":477898540,"identity":"5d36963e-8309-4b54-87bc-35f74eba4f57","order_by":3,"name":"Victor Archibong","email":"","orcid":"","institution":"University of Rwanda","correspondingAuthor":false,"prefix":"","firstName":"Victor","middleName":"","lastName":"Archibong","suffix":""},{"id":477898541,"identity":"359f29c9-98e9-4a92-bc55-e7535ad60582","order_by":4,"name":"Ibrahim Ssengendo","email":"","orcid":"","institution":"Kampala International University - Western Campus","correspondingAuthor":false,"prefix":"","firstName":"Ibrahim","middleName":"","lastName":"Ssengendo","suffix":""}],"badges":[],"createdAt":"2025-04-09 15:23:11","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6413247/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6413247/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":85677865,"identity":"1157f70f-cbc6-4f05-8c8c-e4e7bbe30da2","added_by":"auto","created_at":"2025-06-30 14:46:22","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":259668,"visible":true,"origin":"","legend":"\u003cp\u003eHematoxylin and eosin stain. Magnification X100: Seminiferous tubules (ST) lined inside by seminiferous epithelium (SE) supported by an intact basement membrane. SE is formed of cells arranged in many layers: Sertoli cells and cells of spermatogenic series: spermatogonia, primary spermatocyte, secondary spermatocyte and spermatids (round and elongated). Interstitial tissue (IT) of the testis contains blood vessels, connective tissue and Leydig cells\u003c/p\u003e\n\u003cp\u003eA. Round-oval-shaped ST are densely packed in the testicular parenchyma. Elongated spermatids and spermatozoa fill the lumen of the ST\u003c/p\u003e\n\u003cp\u003eB. The germinal epithelium showed increased thickness. IT appeared pale staining due to the accumulation of lipid droplets in testosterone-producing Leydig cells. Blood vessels (red arrow) could be seen centered in the IT\u003c/p\u003e\n\u003cp\u003eC. Closely packed seminiferous tubules with thick SE. The IT is filled with cells of a foamy appearance. Increased vascular elements in the IT\u003c/p\u003e\n\u003cp\u003eD. Empty ST with thin germinal epithelium and wider lumen. Hyalinized tissue due to increased fibroblast cells was conspicuous, along with congested and dilated blood vessels (yellow arrow). Detached germ cells lined some tubules. Arrested spermatogenesis in some tubules\u003c/p\u003e\n\u003cp\u003eE. Congested and dilated blood vessels with detached ST. The IT appears full and indistinct with signs of hyalinization. Spermatozoa occupy irregular-shaped ST.\u003c/p\u003e\n\u003cp\u003eF. Vascularized interstitium with closely packed ST having a regular-shaped outline. Slight epithelial detachment and increased spermatogenesis\u003c/p\u003e","description":"","filename":"1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6413247/v1/2f32d2674783e2a2715b2855.jpg"},{"id":85677868,"identity":"f7053393-8e08-4a59-9510-2eeba3ce3bee","added_by":"auto","created_at":"2025-06-30 14:46:22","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":305043,"visible":true,"origin":"","legend":"\u003cp\u003ePAS histochemical staining. Magnification X100: A, B, C, E, F showing an apparent strong PAS positive reaction of the basement membrane with Groups C and F being intensely strong. Group D showing weak PAS-positive reaction in the basement membrane (arrows).\u003c/p\u003e","description":"","filename":"2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6413247/v1/0769eea779f6fde7dccf7b8e.jpg"},{"id":85679104,"identity":"9c75194b-5770-477d-9f7f-fb2c5f2b031e","added_by":"auto","created_at":"2025-06-30 14:54:22","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":289913,"visible":true,"origin":"","legend":"\u003cp\u003eMasson’s Trichrome staining of testis. Magnification X100:\u003c/p\u003e\n\u003cp\u003eA, B and C showing minimal collagen fiber deposition in the tunica albuginea (capsule surrounding testis), the IT and ST basement membrane and blood vessels. The deposition (blue) was intensive in groups D and E. F showed the least deposition of collagen fibers in the tunica albuginea, the IT and ST basement membrane, and blood vessels.\u003c/p\u003e","description":"","filename":"3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6413247/v1/3bef0ada3ed1d7f51b5d3d16.jpg"},{"id":103404059,"identity":"af09cd84-2174-450a-acdf-f9baef258d6d","added_by":"auto","created_at":"2026-02-25 09:44:50","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1253622,"visible":true,"origin":"","legend":"","description":"","filename":"FINALMANUSCRIPT230625.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6413247/v1_covered_d780794f-61ff-4d37-97ec-f78d6dc77edb.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Ethanolic seed extract of Artocarpus heterophyllus ameliorates histomorphological, morphometric and functional alterations in the rat testis induced by sildenafil citrate ","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"ackfruit, Artocarpus heterophyllus, sildenafil citrate, male fertility, testicular morphology, serum testosterone, Leydig cells, seminiferous tubules","lastPublishedDoi":"10.21203/rs.3.rs-6413247/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6413247/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground:\u003c/strong\u003e One in every ten men is affected by ED regardless of their age. \u0026nbsp;Sildenafil citrate is the first-line \u0026nbsp;drug \u0026nbsp;in \u0026nbsp;the \u0026nbsp;treatment \u0026nbsp;of \u0026nbsp;ED \u0026nbsp;and \u0026nbsp;has \u0026nbsp;been \u0026nbsp;associated \u0026nbsp;with \u0026nbsp;seminiferous \u0026nbsp;tubules \u0026nbsp;and \u0026nbsp;testicular degeneration, translating into decreased spermatogenesis. The study evaluated the effect of ethanolic seed extract \u0026nbsp;of \u0026nbsp;Artocarpus \u0026nbsp;heterophyllus \u0026nbsp;on \u0026nbsp;changes \u0026nbsp;in \u0026nbsp;testicular \u0026nbsp;histology, \u0026nbsp;seminal \u0026nbsp;fluid \u0026nbsp;and \u0026nbsp;serum testosterone levels alone and in ameliorating testicular changes induced by Sildenafil Citrate.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods:\u003c/strong\u003e Thirty-six adult male Wistar rats were randomly divided into six groups (n=6) and orally dosed daily with NS, seed extract of AH (500 or 1000 mg/kg), SC (50 mg/kg) or AH+SC (1000 mg/kg + 50 mg/kg or 1000 mg/kg + 100 mg/kg). Testicular histopathological and semen analyses were performed via H\u0026amp;E, PAS and Masson’s trichome staining and microscopy. Differences in sperm parameters, testicular size, \u0026nbsp;seminiferous \u0026nbsp;tubule \u0026nbsp;characteristics, \u0026nbsp;Leydig \u0026nbsp;cell \u0026nbsp;volume, \u0026nbsp;and \u0026nbsp;serum \u0026nbsp;testosterone \u0026nbsp;levels \u0026nbsp;were determined.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults: \u003c/strong\u003eSC \u0026nbsp;administration \u0026nbsp;significantly \u0026nbsp;decreased \u0026nbsp;the \u0026nbsp;sperm \u0026nbsp;count \u0026nbsp;(3.47±1.46), \u0026nbsp;motility \u0026nbsp;(23.414±6.63) \u0026nbsp;and morphology (0.038±0.02) compared \u0026nbsp;to the negative control (6.04±0.47, 27.888±0.81 and 0.058±0.02, respectively). The administration of AH 1000 to SC-positive rats improved these parameters in an SC dose-dependent manner, i.e., 10.54±1.48, 31.516±0.99 and 0.0781±0.02 and 9.36±0.36, 27.314±1.45 and 0.049±0.03 for 50 mg/kg and 100 mg/kg SC, respectively. AH improved the diameter of the seminiferous tubules but decreased the height in SC-positive rats [(297.2±21.64) and (289.0±8.60)] and [100.2±5.03 and 96.8±4.20] compared with SC alone (194.0±51.05 and 60.0±15.26, respectively). SC decreased the Leydig cell diameter (6696.05±2525.53), AH 1000 significantly reversed this (8218.53±1291.49) to above those of the negative control (7584.18±816.46). Serum testosterone levels were lower in the SC group (2.53±1.88) than in the normal group (3.28±1.74), but these levels were improved by AH (4.596±0.73 and 5.46±2.55). Histological examination of the testes revealed increased spermatogenesis and Leydig cell proliferation with a higher dose of extract.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion: \u003c/strong\u003e\u0026nbsp;This \u0026nbsp;study \u0026nbsp;revealed \u0026nbsp;that \u0026nbsp;the \u0026nbsp;ethanolic \u0026nbsp;seed \u0026nbsp;extract \u0026nbsp;of \u0026nbsp;A. \u0026nbsp;heterophyllus \u0026nbsp;affects \u0026nbsp;male \u0026nbsp;rat reproductive \u0026nbsp;system \u0026nbsp;function \u0026nbsp;by \u0026nbsp;improving \u0026nbsp;sperm \u0026nbsp;parameters, \u0026nbsp;testicular \u0026nbsp;volume, seminiferous \u0026nbsp;tubule characteristics, Leydig cell volume and serum testosterone levels. AH also improved all these parameters in SC-positive rats in an SC dose-dependent manner.\u003c/p\u003e","manuscriptTitle":"Ethanolic seed extract of Artocarpus heterophyllus ameliorates histomorphological, morphometric and functional alterations in the rat testis induced by sildenafil citrate ","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-06-30 14:46:17","doi":"10.21203/rs.3.rs-6413247/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"a7e386cc-2815-4d6a-8559-30e7b024de76","owner":[],"postedDate":"June 30th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-02-25T09:43:35+00:00","versionOfRecord":[],"versionCreatedAt":"2025-06-30 14:46:17","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6413247","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6413247","identity":"rs-6413247","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}