{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,27]],"date-time":"2026-02-27T05:07:52Z","timestamp":1772168872616,"version":"3.50.1"},"update-to":[{"DOI":"10.12688\/f1000research.124947.1","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2023,8,2]],"date-time":"2023-08-02T00:00:00Z","timestamp":1690934400000}},{"DOI":"10.12688\/f1000research.124947.2","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2023,8,2]],"date-time":"2023-08-02T00:00:00Z","timestamp":1690934400000}}],"reference-count":40,"publisher":"F1000 Research Ltd","license":[{"start":{"date-parts":[[2023,8,2]],"date-time":"2023-08-02T00:00:00Z","timestamp":1690934400000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"University grant, Airlangga University","award":["407 \/UN3.14\/PT\/2020"],"award-info":[{"award-number":["407 \/UN3.14\/PT\/2020"]}]}],"content-domain":{"domain":["f1000research.com"],"crossmark-restriction":false},"short-container-title":["F1000Res"],"abstract":"\n Background:<\/ns4:bold>\n Medicinal plants have been demonstrated to possess various pharmacological effects including anti-hepatitis C virus. Acacia \u00a0mangium is one of the Acacia genus that contain various metabolites. The metaboilites play an\u00a0 important role for antiviral activities. The current study examined the anti-hepatitis C virus (HCV) activities of\u00a0\n Acacia mangium<\/ns4:italic>\n \u00a0extracts in solvents with various polarities and further evaluated the mechanism of action of the extracts on the protein virus and combination treatment models.\n <\/ns4:p>\n \n Methods:<\/ns4:bold>\n Anti-hepatitis C virus activities was conducted with\u00a0\n in vitro<\/ns4:italic>\n \u00a0cells culture of Huh 7it both in a single or combination treatment. Its NS3 protein inhibition was evaluated with western blotting assay.\n <\/ns4:p>\n \n Results:<\/ns4:bold>\n The results revealed the strong anti-HCV activities of the extracts. The 50% inhibition concentrations (IC\n 50<\/ns4:sub>\n s) of the ethanol, n-hexane, dichloromethane and methanol extracts were of 4.6 \u00b1 0.3, 2.9 \u00b1 0.2, 0.2 \u00b1 0.3, and 2.8 \u00b1 0.2 \u03bcg\/mL, respectively, and no cytotoxic effect was detected. These extracts displayed stronger effects than the positive control ribavirin. The mode of action of the ethanol extract was evaluated at 30 \u00b5g\/mL, revealing that the inhibitory effect was stronger on the post-entry step than on the entry step. Western blotting revealed that the extracts decreased NS3 protein expression, indicating that virus replication was suppressed. Further evaluation illustrated that combined treatment with the ethanol extract enhanced the anti-viral activity of simeprevir.\n <\/ns4:p>\n \n Conclusions:<\/ns4:bold>\n These results indicated that\n A. mangium<\/ns4:italic>\n leaves could represent sources of anti-HCV agents.\n <\/ns4:p>","DOI":"10.12688\/f1000research.124947.3","type":"journal-article","created":{"date-parts":[[2023,8,2]],"date-time":"2023-08-02T06:05:07Z","timestamp":1690956307000},"page":"1452","update-policy":"https:\/\/doi.org\/10.12688\/f1000research.crossmark-policy","source":"Crossref","is-referenced-by-count":2,"title":["Acacia mangium: A promising plant for isolating anti-hepatitis C virus agents"],"prefix":"10.12688","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2049-7774","authenticated-orcid":false,"given":"Tutik Sri","family":"Wahyuni","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nida S.","family":"Sukma","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7604-2053","authenticated-orcid":false,"given":"Adita A.","family":"Permanasari","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chie","family":"Aoki-Utsubo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4639-1599","authenticated-orcid":false,"given":"Aty","family":"Widyawaruyanti","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0570-7975","authenticated-orcid":false,"given":"Achmad Fuad","family":"Hafid","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"2560","published-online":{"date-parts":[[2023,8,2]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"180-187","DOI":"10.1111\/1348-0421.12127","article-title":"Anti-hepatitis C virus compounds obtained from Glycyrrhiza uralensis and other Glycyrrhiza species.","volume":"58","author":"M Adianti","year":"2014","journal-title":"Microbiol. 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Other revision was also mentioning the bioactivities information of \nAcacia mangium<\/i> and other Acacia genus in the introduction section.  Other comments were regarding the HPLC profile of each extracts that have differences polarities. Not much information regarding this data, however some information of \nA. mangium<\/i> compounds and other \nAcacia <\/i>genus have been explained in the text.  Comment regarding the examination of difference batch of extract have been done and gave the relatively the same activity.  Best regards, Tutik Sri wahyuni, PhD.","URL":"https:\/\/f1000research.com\/articles\/11-1452\/v3#referee-comment-9385","order":1,"name":"referee-comment-9385","label":"Referee Comment","group":{"name":"article-reports","label":"Article Reports"}},{"value":"10.5256\/f1000research.144860.r170379, Courage Sedem Dzah, Ho Technical University, Ho, Volta Region, 19 May 2023, version 2, 1 approved, 1 approved with reservations, 1 not approved","URL":"https:\/\/f1000research.com\/articles\/11-1452\/v2#referee-response-170379","order":2,"name":"referee-response-170379","label":"Referee Report","group":{"name":"article-reports","label":"Article Reports"}},{"value":"tutik sri wahyuni<\/b>; \nPosted: 01 Jun 2023<\/i>; Dear Reviewer,  Thank you for your comment and suggestion.  Here with some point of our responses:  1. "...\nAcacia mangium <\/i>is one of the \nAcacia <\/i>genus that contain \nwith<\/u> various metabolites..." - Take out the "with" I have underlined. \nAnswer<\/b>: We have revised the manuscript and remove the word \u201dwith\u201d 2. "The current study examined the anti-hepatitis C virus (HCV) activities of \nAcacia mangium<\/i> extracts in solvents with various polarities and further evaluated the mechanism of action of the extracts on t\nhe protein virus and combination treatment models<\/u>." - Are you evaluating effects on combination treatment models too or on protein virus? I do not think the models are also dependent variables.  \nAnswer<\/b>: Our study evaluated the effect of extract in the NS3 protein of virus. And it demonstrated the inhibition effect of extract in NS3 protein level of virus. Furthermore, the combination analysis was conducted between extract and simeprevir which showed enhancement effects of extract on anti-HCV activity of simeprevir. Those results suggested that extract has a role in protein inhibition. However, our study did not examine directly the mixture of extract and simeprevir on protein virus. 3. "Anti-hepatitis C virus activities was conducted by \nin vitro<\/i> culture cells..." - Please replace "by" with "using". Because culture cells cannot conduct a test. Also, better to say cell culture than "culture cell". \nAnswer<\/b>: Thank you for the suggestion. We have revised in the manuscript, changed the word 'by' with 'using' and changed 'culture cells' with 'cell culture'.    4."Further \nexamined<\/u> its NS3 protein inhibition was evaluated \nby<\/u> western blotting assay..." - This whole statement is grammatically incorrect. Take out "examined" and replace "by" with "using". \nAnswer<\/b>: We have revised in the manuscript   5. What is ribavirin? Any brief details on it and why was it used as a control?  Answer: Ribavirin was used as a treatment in hepatitis C virus infection. It has several mechanism, (1) Modulate the host immunity by inferfare the T helper 1 and T helper 2, (2) Inhibit the inosine monophosphate dehydrogenase and depletion of guanosine triphosphate which resulting the inhibition of viral replication and protein synthesis, (3) ribavirin also act as mutagen in the viral target. Although the Direct Acting Antiviral Agents have been used as current treatment of hepatitis C virus, however some developing area still used ribavirin and specific patient condition was need with ribavirin combination treatment. 6. "The mode of action of the ethanol extract was evaluated at 30 µg\/mL, revealing that the inhibitory effect was stronger on the post-entry step than on the entry step." - what does entry and post-entry steps mean in this context and why study kinetics at a higher concentration than the 4.6 ug\/ml for ethanol extract? \nAnswer<\/b>: The Hepatitis C Virus life cycle is mainly divided into two stage; entry step (viral adsorption and penetration) and post-entry step (uncoating, translation, replication, assembly and release). The time-of-addition study is performed to determine antiviral effect of the plant extract in the HCV life cycle, and the plant extract concentrations which exhibits >90% virus inhibition (30 µg\/mL in this experiment) is usually used to analyse. 7. Why use ribavirin as a positive control and also use simeprevir in combination treatment? Comparing simeprevir-based treatment with the control will not make sense.  \nAnswer<\/b>: The purpose using ribavirin as positive control not for comparing with simeprevir. We use simeprevir as a combiantion treatment due to the mechanism of simeprevir as NS3 protein inhibitor.  8. "...The current study examined the anti-hepatitis C virus (HCV) activities of \nAcacia mangium<\/i> extracts in solvents with various polarities..." - Please indicate here early enough that you used the leaves of the plant. This was not mentioned until your conclusion and it should not be so.  \nAnswer<\/b>: We have revised by added leave in the introduction section. We have mentioned in the material section that leaves were used of \nAcacia mangium<\/i> extract. 9. Why extract compounds from 200g of leaf powder? What informed this figure?  \nAnswer<\/b>: Plant material preparation was derived from the powder of dried leaves. A number of 200 g was used for extraction process with ethanol solvent and other 200 g of \nA. mangium<\/i> powder was extracted with difference polarities solvent, n-hexane, dichloromethane and methanol). 200 g of sample is enough for the research purpose. 10. At what temperature was the extraction done? \nAnswer<\/b>: The extraction was done in the room temperature.   11. For how long was the maceration done? \nAnswer<\/b>: The maceration was done overnight in three times.   12. For your MTT assay, why did you not rinse your cells and take out media and add DMSO before incubating again for colour development for spectral reading? DMSO is added to MTT assays at the end of the experiment to dissolve the formazan crystals that were created during the experiment. If DMSO is not added and the formazan crystals are not dissolved, this would cause significant changes in the absorbance spectrum due to the sodium bicarbonate that would still be there.  \nAnswer<\/b>: The MTT assay was conducted as you was mentioned. We have revised in the manuscript. After 2 days incubation, the medium was discarded, rinse with medium then 150 mikroliter of 10% of MTT reagent was added and incubate for 4 hours. After incubation time removed the MTT and DMSO was added to dissolve the formazon. 13. How sure are you that you have removed interference from cell culture media and that your result is not as a result of potential interference? \nAnswer<\/b>: The cytotoxic assay was done following our previous method and replication was also conducted to ensure the valid result. Other concerns 14. What compounds from the plant leaf specifically are responsible for the results you have gotten? \nAnswer<\/b>: Our study have not deermine the single compounds with responsible to the activities. Following to the references epicatechin, quercetin, proacaciaside I, and proacaciaside II were reported as isolated compounds of Acacia genus that may contribute in their activities.  15. Figure 5 throws light on 5 bioactive compounds. Are the observations as a result of all of these compounds or some of them or just one of them? \nAnswer<\/b>: The compounds in Figure 5 were known as isolated compounds of Acacia genus that may give contribution in its activities. 16. Does it mean that the results are only due to these five compounds or there may be other compounds beyond these five that may give the results you have? \nAnswer<\/b>: Our studies have not determined the specific compounds with contribute for its anti-HCV activities. There are strill many unidentified compounds from this plants. 17. Please, do consider English Language Editing to avoid some errors.   \nAnswer<\/b>: We have done the proof read editing and revised some portion. Thank you","URL":"https:\/\/f1000research.com\/articles\/11-1452\/v3#referee-comment-9730","order":3,"name":"referee-comment-9730","label":"Referee Comment","group":{"name":"article-reports","label":"Article Reports"}},{"value":"10.5256\/f1000research.144860.r172677, Borris Rosnay T Galani, Universite de Ngaoundere, Ngaoundere, Adamawa, 05 Jun 2023, version 2, 1 approved, 1 approved with reservations, 1 not approved","URL":"https:\/\/f1000research.com\/articles\/11-1452\/v2#referee-response-172677","order":4,"name":"referee-response-172677","label":"Referee Report","group":{"name":"article-reports","label":"Article Reports"}},{"value":"tutik sri wahyuni<\/b>; \nPosted: 28 Jul 2023<\/i>; Dear Reviewer  Thank you for your comments and suggestions. Herewith our respond to the reviewer comments Introduction: Many references in the introduction are quite old, for example, Swain et al. 2010; Javed et al. 2011. The literature needs to be updated (2018-2023). \nAnswer: <\/b>We revised some references. Javed was change to Segarra-Newnham M, See N, Fox-Seaman G. Retreatment of Hepatitis C Infection With Direct-Acting Antivirals. Fed Pract. 2020 Jul;37(7):316-319. PMID: 32908335; PMCID: PMC7473721.   New efforts also result in the approval of direct-acting antivirals, or DAAs, which aid in the recovery from HCV in three months. Sofosbuvir, Daclatasvir, and other DAAs are quite effective. The major restriction here is the emergence of resistance to these new medications. To validate the title of this article, I would recommend that the authors evaluate the antiviral effects of the plant extracts on HCV-resistant strains. \nAnswer:<\/b> Thanks for the recommendation. However, our main concern here is to show the potential of the \nAcacia mangium<\/i> plant as a complementary drug against HCV. Therefore, we are also conducting combination treatment with simaprevir. Please write the species names in italics and correct spelling errors: Acacia nilotica, not 'Acacia nilotoca'. \nAnswer:<\/b> The spelling and grammar errors have been proofread by a native English speaker and have been revised as per your suggestions. Simeprevir is an NS3\/4A protease inhibitor used to treat HCV-genotypes 1 and 4 patients. Since the authors used this drug as a positive control in their tests, actual data on this compound needs to be presented in the introduction as well as its major limitation. Strengths of Simeprevir: High efficacy: Simeprevir has demonstrated high rates of sustained virologic response (SVR) in clinical trials, leading to viral clearance and improved patient outcomes. Genotype specificity: It is particularly effective against HCV genotypes 1 and 4, making it a valuable treatment option for patients infected with these genotypes. Once-daily dosing: Simeprevir is typically administered as a once-daily oral medication, which improves convenience and patient adherence to the treatment regimen. Weaknesses of Simeprevir: Resistance development: The long-term use of Simeprevir, especially as a monotherapy, can lead to the development of viral resistance, potentially reducing its effectiveness. Limited efficacy in certain populations: Simeprevir may not be as effective in certain patient populations, such as those with liver cirrhosis or prior treatment failure. Drug interactions: Simeprevir has the potential for drug interactions, particularly with certain medications metabolized by cytochrome P450 enzymes, which may require dosage adjustments or careful monitoring. Reference: Rice CM, et al. (2014). "Simeprevir - a protease inhibitor for the treatment of hepatitis C virus infection." N Engl J Med, 370(8): 796-797. Why did the authors confine the study to in vitro systems rather than including an in vivo examination? summaries of the opportunities provided by in vitro systems must be presented here. \nAnswer: <\/b>The study of in vitro systems refers to conducting experiments or investigations in a controlled laboratory environment outside of a living organism. In vitro studies involve using cells, tissues, or organs cultured in a laboratory setting, often in artificial conditions that mimic aspects of the natural environment. And for HCV the in vivo model was very limited Methodology: Please also include the length of the maceration of plant extracts. \nAnswer:<\/b> The maceration process has been described Please properly cite Wahyuni, 2013 and include it in the references. But this reference is also old. \nAnswer<\/b>: The appropriate method was described very well in Wahyuni 2013. And the method was used the same until now. The Huh7it cell systems must be introduced correctly. What makes them superior to other JFH-1 systems? \nAnswer:<\/b> The Huh7 cell system, is a widely used in vitro cell culture model for studying the hepatitis C virus (HCV). It is derived from the Huh-7 human hepatoma cell line and has been extensively employed for HCV research, including the study of the JFH-1 strain of the virus. Two positive controls (ribavirin and simeprevir) were employed independently in this study with no evidence of their mode of action. To minimize confusion, this must be described explicitly. Also, clarify why Sofosbuvir a very popular replication inhibitor has not been used in this assessment. It would have helped to understand the extent of this effect on the replication step. \nAnswer:<\/b> We used ribavirin and simeprevir that still used as anti-hcv drug in some area in developing countries due to limited others DAAs For the suppression of HCV infectivity under the section "Anti-HCV activity," I would propose that the authors specify which viral proteins the primary (human) antibody employed for immunostaining precisely targets, and that they solely refer to prior publications (Permanasari & Wahyuni, 2022). \nAnswer: <\/b>We used the serum of a patient with a positive HCV genotype 2a, which did not have a specific viral protein target. I believe that at least one mean comparison test should be used to detect the statistical difference between IC\n50<\/sub> values. \nAnswer: <\/b>We complete it with Student's t-test: The t-test is a parametric statistical test that compares the means of two groups. Since we have two sets of IC\n50 <\/sub>values   What about the isobologram curves now that combination experiments have been completed? Before concluding the Simeprevir relationship, it would have been great to know whether this combination is synergistic or antagonistic. \nAnswer:<\/b> We have completed with the diagram which showed synergetic effect by Combination Index (CI) analysis through compusyn software. The Combination Index is a quantitative measure used to evaluate drug interactions in combination therapies. It is calculated based on the dose-response curves of individual drugs and the combination. The CI value assesses whether the observed effect of the combination is synergistic (CI < 1), additive (CI = 1), or antagonistic (CI > 1). Results & Discussion According to my interpretation of Table 1, the dichloromethane extract has the lowest IC50 value and the highest selectivity index (SI), and therefore should be used for further analysis. I'm curious why the authors chose to focus on the ethanol extract instead. Please stick to facts. \nAnswer:<\/b> The author was more explored in ethanol extract because of the potency in developing extanol extract as herbal drug product. Based on this result, I think the phytochemical characterization of the dichloromethane extract by HPLC\/UV is necessary. Kindly support these findings with in silico studies of Acacia mangium compounds with the HCV NS3\/4A protease. \nAnswer:<\/b> We will conduct it on the future experiments   The information in Figure 1 and Table 1 is almost the same. I would advise the authors to concentrate solely on Table 1 because it includes IC50, CC50, and SI values, which are more relevant. Because IC50 is a result of a dose-dependent impact, showing it directly explains it and eliminates the need for Fig. 1. \nAnswer: <\/b>The figure 1 we described the doses dependent manner among concentration and percent inhibition. While in the table represent the IC50, CC50 and SI, so we think it also necessary to include figure 1.   Fig 5: Honestly, I don't see the importance of these compounds since you have not isolated all these from your plant. In my opinion, you must isolate these compounds and test their effect on replication. \nAnswer:<\/b> The five compounds were active compounds that showed potential as both antibacterial and antiviral agents since we did not perform the isolation. This information will be very useful for predicting compounds that may play as anti-HCV agents. We agree that more study was needed to elucidate the compounds and determine their activities. What about the effects of these extracts on other non-structural proteins including the RdRp which is a key enzyme in the replication step? \nAnswer:<\/b> We not perform the PCR for RdRp. Our study was detected the NS3 protein level, that also a key component of the hepatitis C virus (HCV) replication. It plays essential roles in viral replication and is considered an attractive target for antiviral drug development. The effect of this extract on NS3\/4A activity is also required We agree with the reviewer suggestion, however some limiting of experiment condition that could not examine that matter Conclusion Before concluding that\n A. mangium<\/i> extracts might be exploited to produce supplementary and alternative therapies for HCV more in vivo research is needed to evaluate the anti-HCV potential and assess the toxicity. \nAnswer:<\/b> Yes, sure. However, our concern was to analyze the activity of Acacia mangium in inhibiting HCV growth in an in vitro model. The in vivo study will be conducted the the future research.","URL":"https:\/\/f1000research.com\/articles\/11-1452\/v3#referee-comment-9983","order":5,"name":"referee-comment-9983","label":"Referee Comment","group":{"name":"article-reports","label":"Article Reports"}},{"value":"10.5256\/f1000research.150283.r222958, Moustafa Sarhan, Zoology Department, Faculty of Science, Al-Azhar University, Assiut, 30 Nov 2023, version 3, indexed","URL":"https:\/\/f1000research.com\/articles\/11-1452\/v3#referee-response-222958","order":6,"name":"referee-response-222958","label":"Referee Report","group":{"name":"article-reports","label":"Article Reports"}},{"value":"tutik sri wahyuni<\/b>; \nPosted: 05 Jun 2024<\/i>; Dear Reviewer, Thank you for your comment and sugession. Regarding the title, we would like to express that this results provide the promissing candicate for anti-HCV of Acacia mangium We have provide the Fig 1 and Fig 2 with high resolution figures. We suggest to download the figure so it can access clearly. In the mode of action we used three pararel assay. Evaluate the effect in during inoculation (sample and virus), post inoculation\/post entry and whole step.  The treatment of cell with extract only is suitable for evaluate pre entry\/ before virus infection that suitable for prevention evaluation assay. We do further analyze for ethanol extract because ethanol extract less toxic and it is a recommendation solvent in product development. We have on going process for further examine the difference profile among the extract and purification to obtain the active compounds. Thank you in advance ","URL":"https:\/\/f1000research.com\/articles\/11-1452\/v3#referee-comment-11719","order":7,"name":"referee-comment-11719","label":"Referee Comment","group":{"name":"article-reports","label":"Article Reports"}},{"value":"This research was supported by the internal university grant of Universitas Airlangga with the grant number 407 \/UN3.14\/PT\/2020. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.","order":8,"name":"grant-information","label":"Grant Information"},{"value":"This is an open access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.","order":0,"name":"copyright-info","label":"Copyright"}]}}