PONE-D-24-11983Resveratrol attenuates non-steroidal anti-inflammatory drug-induced intestinal injury in rats in a high-altitude hypoxic environment by modulating the TLR4/NFκB/IκB pathway and gut microbiota compositionPLOS ONE

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PLOS ONE

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Dear Authors,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE publication criteria as it currently stands. The shortcomings of this paper needs to be worked out before it can be considered for publication. Therefore, we invite you to resubmit a revised version of the manuscript that addresses the points raised during the review process.

For your guidance, the reviewers' comments are included below.

Thank you for giving us the opportunity to consider your work.

Specific concerns expressed during peer review were:

Comments from Reviewer 1

Dear Dr. Feng Gao and colleagues,

Your manuscript titled "Resveratrol attenuates non-steroidal anti-inflammatory drug-induced intestinal injury in rats in a high-altitude hypoxic environment by modulating the TLR4/NFκB/IκB pathway and gut microbiota composition" has been reviewed. The study is of significant interest, addressing the protective role of resveratrol against NSAID-induced intestinal damage in a high-altitude setting. However, there are several areas that require major revisions to enhance the clarity, depth, and impact of the manuscript. Please find the detailed comments below.

Introduction and Background (Section 1):

The linkage between NSAID usage and intestinal damage in high-altitude environments is well-noted but could be strengthened with recent literature to underline the relevance and timeliness of the study.

Please expand on the molecular mechanisms of the TLR4/NFκB/IκB pathway as it relates to intestinal inflammation, providing a clearer background for readers less familiar with this area.

Methods (Sections 2.1 to 2.9):

In section 2.3, the experimental design should explicitly mention any ethical considerations related to the handling and treatment of the animals beyond the approval by the ethics committee.

Please provide more details on the blinding process during the experimental assessments to avoid bias, especially during the injury scoring and histopathological examinations (Section 2.4, 2.5).

The statistical methods section (2.9) could benefit from additional details regarding the assumptions checked before the analysis and the rationale behind choosing specific statistical tests.

Results (Section 3):

In section 3.2 and 3.3, please provide the exact p-values rather than just stating statistical significance. This would add to the transparency and reproducibility of the results.

The Western blot images and corresponding densitometry data (Section 3.5) should be included in the supplementary files if not in the main manuscript, for validation of the protein expression data reported.

Discussion (Section 5):

The discussion could be improved by contrasting your findings with existing studies, particularly those that have not observed beneficial effects of resveratrol or have different findings regarding gut microbiota changes under similar conditions.

Please address potential limitations of the study, such as the translation of rat model findings to human conditions and the implications of the doses of resveratrol used.

Figures and Tables:

Ensure that all figures are of high quality and appropriately labeled for clarity. Figure legends should be descriptive enough to be understood independently of the main text.

Tables summarizing the raw data and statistical analysis should be provided to enhance the clarity and depth of the reported findings.

References:

Several references appear outdated. Please update these citations with more current research articles that reflect the latest developments in the field.

Ethical Statement:

The ethical statement should be more detailed, especially regarding the specific steps taken to minimize animal suffering and the rationale behind the number of animals used.

Comments from Reviewer 2

Title

•“TLR4/NFκB/IκB pathway”, if all these pathways are involved then why is “/” used? / indicate one of these three pathways. Please clarify the situation

Abstract:

•Line 3, gastrointestinal tract can be abbreviated as GIT and later use the abbreviation instead of the full form.

•ZO-1, first time use the full form then use the abbreviated form.

•RSV, avoid using abbreviations at the start of sentences.

Background:

•In line 5, use NSAIDs, instead of full from “nonsteroidal anti-inflammatory drugs”. There is no consistency in the article. Please check the whole manuscript.

•Rsv, check this for capitalization, in all other places RSV is used. Be consistent.

•More related information can be added in the introduced to make the mechanism of action of given treatment clearer (Resveratrol)

Methodology:

•Experimental design can be represented in the form of a flow chart, or in tabulated form.

Results:

3.2 HE staining results:

•In 2nd last line “mitigates” should be replaced by a suitable word like alleviates. As it is not suitable to use this word here.

3.4:

•RSV can effectively reduce intestinal damage caused by taking nonsteroidal anti-inflammatory drugs under high-altitude hypoxia conditions”, here rearrange this heading, especially “by taking” can be replaced by intake. And use NSAIDs, instead of the full form. Throughout check such types of things, the main headings and subheadings should not be too lengthy.

4.2 Bacterial diversity:

•signifying significantly, check this repetition doesn’t seem attractive.

4.3Microbial composition:

•Clostridium_sensu_stricto, check if sensu_stricto should be italicized according to scientific nomenclature. Apply the same rule to all bacterial phylum and genera, to check the rule for their italicization as above mentioned (for example Clostridiales_Incertae_sedis_XI, Ruminococcus_callidus, etc)

Discussion:

•MPO, avoid abbreviations at the start of a sentence.

Clarity and Structure:

•The clear mechanism between the high altitude, low oxygen pressure, and intestinal damage is not mentioned in the discussion, or in the introduction section

Overall Impression:

•Need to be grammar-checked, spell spell-checked by an expert who is fluent in English.

Recommendations:

•Need to be revised again strictly, by an expert in the given field.

Please submit your revised manuscript by 30th May, 2024. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

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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

**********

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

Reviewer #1: No

Reviewer #2: Yes

**********

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: No

Reviewer #2: Yes

**********

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

**********

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: Dear Dr. Feng Gao and colleagues,

Your manuscript titled "Resveratrol attenuates non-steroidal anti-inflammatory drug-induced intestinal injury in rats in a high-altitude hypoxic environment by modulating the TLR4/NFκB/IκB pathway and gut microbiota composition" has been reviewed. The study is of significant interest, addressing the protective role of resveratrol against NSAID-induced intestinal damage in a high-altitude setting. However, there are several areas that require major revisions to enhance the clarity, depth, and impact of the manuscript. Please find the detailed comments below.

Introduction and Background (Section 1):

The linkage between NSAID usage and intestinal damage in high-altitude environments is well-noted but could be strengthened with recent literature to underline the relevance and timeliness of the study.

Please expand on the molecular mechanisms of the TLR4/NFκB/IκB pathway as it relates to intestinal inflammation, providing a clearer background for readers less familiar with this area.

Methods (Sections 2.1 to 2.9):

In section 2.3, the experimental design should explicitly mention any ethical considerations related to the handling and treatment of the animals beyond the approval by the ethics committee.

Please provide more details on the blinding process during the experimental assessments to avoid bias, especially during the injury scoring and histopathological examinations (Section 2.4, 2.5).

The statistical methods section (2.9) could benefit from additional details regarding the assumptions checked before the analysis and the rationale behind choosing specific statistical tests.

Results (Section 3):

In section 3.2 and 3.3, please provide the exact p-values rather than just stating statistical significance. This would add to the transparency and reproducibility of the results.

The Western blot images and corresponding densitometry data (Section 3.5) should be included in the supplementary files if not in the main manuscript, for validation of the protein expression data reported.

Discussion (Section 5):

The discussion could be improved by contrasting your findings with existing studies, particularly those that have not observed beneficial effects of resveratrol or have different findings regarding gut microbiota changes under similar conditions.

Please address potential limitations of the study, such as the translation of rat model findings to human conditions and the implications of the doses of resveratrol used.

Figures and Tables:

Ensure that all figures are of high quality and appropriately labeled for clarity. Figure legends should be descriptive enough to be understood independently of the main text.

Tables summarizing the raw data and statistical analysis should be provided to enhance the clarity and depth of the reported findings.

References:

Several references appear outdated. Please update these citations with more current research articles that reflect the latest developments in the field.

Ethical Statement:

The ethical statement should be more detailed, especially regarding the specific steps taken to minimize animal suffering and the rationale behind the number of animals used.

Reviewer #2: Title

�“TLR4/NFκB/IκB pathway”, if all these pathways are involved then why is “/” used? / indicate one of these three pathways. Please clarify the situation

Abstract:

�Line 3, gastrointestinal tract can be abbreviated as GIT and later use the abbreviation instead of the full form.

�ZO-1, first time use the full form then use the abbreviated form.

�RSV, avoid using abbreviations at the start of sentences.

Background:

�In line 5, use NSAIDs, instead of full from “nonsteroidal anti-inflammatory drugs”. There is no consistency in the article. Please check the whole manuscript.

�Rsv, check this for capitalization, in all other places RSV is used. Be consistent.

�More related information can be added in the introduced to make the mechanism of action of given treatment clearer (Resveratrol)

Methodology:

�Experimental design can be represented in the form of a flow chart, or in tabulated form.

Results:

3.2 HE staining results:

�In 2nd last line “mitigates” should be replaced by a suitable word like alleviates. As it is not suitable to use this word here.

3.4:

�RSV can effectively reduce intestinal damage caused by taking nonsteroidal anti-inflammatory drugs under high-altitude hypoxia conditions”, here rearrange this heading, especially “by taking” can be replaced by intake. And use NSAIDs, instead of the full form. Throughout check such types of things, the main headings and subheadings should not be too lengthy.

4.2 Bacterial diversity:

�signifying significantly, check this repetition doesn’t seem attractive.

4.3Microbial composition:

�Clostridium_sensu_stricto, check if sensu_stricto should be italicized according to scientific nomenclature. Apply the same rule to all bacterial phylum and genera, to check the rule for their italicization as above mentioned (for example Clostridiales_Incertae_sedis_XI, Ruminococcus_callidus, etc)

Discussion:

�MPO, avoid abbreviations at the start of a sentence.

Clarity and Structure:

�The clear mechanism between the high altitude, low oxygen pressure, and intestinal damage is not mentioned in the discussion, or in the introduction section

Overall Impression:

�Need to be grammar-checked, spell spell-checked by an expert who is fluent in English.

Recommendations:

�Need to be revised again strictly, by an expert in the given field.

**********

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Reviewer #1: Yes: Dr. Prasad Andhare

Reviewer #2: No

**********

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Resveratrol attenuates non-steroidal anti-inflammatory drug-induced intestinal injury in rats in a high-altitude hypoxic environment by modulating the TLR4/NFκB/IκB pathway and gut microbiota composition

PONE-D-24-11983R1

Dear Dr. Feng Gao,

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.

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,

Palash Mandal

Academic Editor

PLOS ONE