PONE-D-21-00075

Protective role of resolvin D1, a pro-resolving lipid mediator, in nonsteroidal anti-inflammatory drug-induced small intestinal damage

PLOS ONE

Dear Dr. Tanigawa

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Hiroyasu Nakano, M.D., Ph.D.

Academic Editor

PLOS ONE

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- https://www.gastrojournal.org/article/S0016-5085(20)32578-6/pdf?referrer=https%3A%2F%2Fapi.ithenticate.com%2F

- https://www.intechopen.com/books/municipal-solid-waste-management/life-cycle-inventory-lci-modeling-of-municipal-solid-waste-msw-management-systems-in-kosodrza-commun

https://www.nature.com/articles/mi201589?code=4eb62728-cfde-4eba-97dd-898b4180c86b&error=cookies_not_supported

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Additional Editor Comments:

Although both reviewers feel the study is potentially interesting, one of the reviewers claims a flaw of the authors’ experimental conditions. Specifically, the reviewer strongly recommends the authors to collaborate with lipid biochemists to correctly measure lipid mediators including resolving D1 in their samples.

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Partly

Reviewer #2: Yes

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2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

Reviewer #2: Yes

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3. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: Yes

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4. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: Yes

Reviewer #2: Yes

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5. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: Kuzumoto et al showed the resolving D1, an anti-inflammatory lipid mediator derived from DHA ameliorated the NSAIDs-induced intestinal damage by inhibiting the intestinal inflammation. An inhibitor for 12/15-lipoxygenase, a critical enzyme for resolving D1 synthesis, increased the expression of inflammatory cytokines and exacerbated the intestinal damage, which was again ameliorated by exogenous resolving D1. Most of the experiments and their results are nicely organized and discussed, except for the quantification of resolving D1 by Elisa.

Major comments

In figure 3a-d, they showed the marked increase in the lesion index, histological score and the expression of inflammatory cytokines by baicalein, a 12/15 lipoxygenase inhibitor, however, they showed the minimal reduction of resolving D1 by baicalein treatment in Figure 3e. This small change in resolving D1 can not explain the huge difference in the clinical scores and inflammation. This is possibly due to the inadequate measurement of resolving D1. They extracted fatty acid fraction from intestinal tissues by methanol and quantified resolving D1 using an Elisa kit, which does not give the reliable values. Currently, lipid mediators including SPM2 should be measured by mass-spectrometry as proposed by Bo Burla et al. in J. Lipid Res. 2018;59:2001-2017. The reviewer strongly recommends the authors to collaborate with lipid biochemists to correctly measure lipid mediators including resolving D1 in their samples.

Reviewer #2: In the manuscript entitled "Protective role of resolvin D1, a pro-resolving lipid mediator, in nonsteroidal anti-inflammatory drug-induced small intestinal damage" by Kuzumoto et al, the authors demonstrate that administration of resolvin D1 ameliorates NSAIDs-induced damages of the small intestine in mice (Fig. 1). The expression of the resolvin D1-producing enzyme (12/15-lipoxygenase) is not affected after administration of indomethacin (Fig. 2). The indomethacin-induced intestinal damages are aggravated in mice that are pre-treated with the inhibitor of 12/15-lipoxygenase, and the aggravation is abrogated by exogenous resolvin D1 (Fig. 3). The indomethacin-induced intestinal damages are also aggravated by the inhibitors of a resolvin D1 receptor (ALX1/FPR2), which is mainly expressed in macrophages (Fig. 4 and Fig. 5).

I feel that the present study provides important findings regarding the role of the endogenous lipid mediator resolvin D1 in regulation of inflammation, and thus seems to be suitable for publication if the following minor issues are adequately addressed.

Specific comments:

1) The authors should measure the concentration of resolvin D1 in the small intestine before and after administration of indomethacin, while they have shown that the expression of 12/15-lipolyxegease is not affected by the administration. Alternatively, appropriate references should be cited in terms of the alteration of the enzymatic activity during the inflammatory responses.

2) The scale bar should be included in the right panel of Fig. 2a.

3) In Figs. 1b and 1e, the time point at the observation after indomethacin administration should be presented (possibly 24 h).

4) For all experiments in Fig. 3, the timing and duration of pre-treatment of baicalein should be provided.

5) The gene names should be formally written for Il1b and Tnf (both in italic), instead of those in the current manuscript.

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Reviewer #1: No

Reviewer #2: No

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Protective role of resolvin D1, a pro-resolving lipid mediator, in nonsteroidal anti-inflammatory drug-induced small intestinal damage

PONE-D-21-00075R1

Dear Dr. Tanigawa

We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.

Although one reviewer still has a concern about the quantification of the indicated lipids of mass spectrometry, I have agreed with the authors’ response that the quantification did not work well at least under the authors’ experimental conditions. Thus, I have accepted the manuscript.

Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication.

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Kind regards,

Hiroyasu Nakano, M.D., Ph.D.

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #1: (No Response)

Reviewer #2: All comments have been addressed

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2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Yes

Reviewer #2: (No Response)

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3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

Reviewer #2: (No Response)

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4. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: (No Response)

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5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: Yes

Reviewer #2: (No Response)

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6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: As a lipid biochemist, the reviewer can not accept the results drawn by SPM measurements by ELISA, even though some papers using this ELISA have been published.

Reviewer #2: (No Response)

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7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #1: No

Reviewer #2: No