PONE-D-23-42843The impact of Astragaloside IV on the inflammatory response and Gut microbiota in cases of acute lung injury is examined through the utilization of the PI3K/AKT/mTOR pathway.PLOS ONE

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Additional Editor Comments:

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:

Editor’s comments:

This manuscript addresses a timely topic and makes a relevant contribution to the field which elaborates how AS-IV reduces the expression of inflammatory factors by inhibiting the PI3K/AKT/mTOR pathway and optimizes the composition of the gut microflora in AIL rats. The author could have been explored the possibility of potential involvement of other potential molecular mechanistic pathways like MAPK, ERK or JAK-STAT pathways.

Comments from Reviewer 1

Dear Authors,

Your manuscript titled "The impact of Astragaloside IV on the inflammatory response and Gut microbiota in cases of acute lung injury examined through the PI3K/AKT/mTOR pathway" presents a comprehensive study exploring the therapeutic potential of Astragaloside IV (AS-IV) in the context of acute lung injury (ALI) and its effect on gut microbiota through modulation of the PI3K/AKT/mTOR pathway. The study is well-structured, and the methodology is robust, covering a wide range of techniques from histological analysis to gene expression and microbiota composition analysis. However, there are several areas where improvements could enhance the clarity and impact of your findings:

1. Literature Review: While your introduction provides a good background, it would be beneficial to further discuss previous studies that have explored similar treatments for ALI, particularly focusing on those involving gut microbiota modulation. This would contextualize your findings within the broader research landscape.

2. Methodological Details: In several sections, the methodology could benefit from more detailed descriptions to ensure reproducibility. For example, specifics regarding the conditions under which the 16S rRNA sequencing was performed (such as sequencing depth and bioinformatics tools used) would be valuable.

3. Statistical Analysis: While you have employed appropriate statistical tests, the manuscript would benefit from a more detailed explanation of the choice of these tests and any assumptions made. Additionally, include information on the corrections applied for multiple comparisons, if any.

4. Results Interpretation: The results are compelling but could be further strengthened by a more detailed discussion of the implications of your findings, particularly the mechanistic insights into how AS-IV modulates the gut-lung axis. Consider discussing potential limitations of the PI3K/AKT/mTOR pathway's role in mediating these effects.

5. Figures and Tables: Ensure that all figures and tables are clearly labeled and referenced in the text. Some figures, such as the graphical abstract and histological images, could benefit from higher resolution or more detailed annotations to aid interpretation.

6. Conclusion and Future Directions: The conclusion succinctly summarizes the findings but could be expanded to more explicitly state the potential clinical implications of your research and suggest specific areas for future studies.

7. References: Check the currentness of your references, especially focusing on recent advances that might complement your discussion.

8. Ethical Considerations: Your ethical approval statement is in order, but please ensure that any additional ethical considerations, particularly concerning animal welfare, are comprehensively addressed.

Comments from Reviewer 2

The present manuscript entitled 'The impact of Astragaloside IV on the inflammatory response and Gut microbiota in

cases of acute lung injury is examined through the utilization of the PI3K/AKT/mTOR

pathway.' describes efficacy of a natural triterpenoid saponin compound, Astragaloside IV(AS-IV), in the treatment of AIL by the suppression of the inflammatory factors through

PI3K/AKT/mTOR pathway and optimizes the composition of the gut microflora in AIL

rats. The study has several shortcomings though is of scientific merit.

Q1. What was the rationale of selecting the dose and duration of AS-IV? What was the route of injection of AS-IV?

Q2. What is the rationale for selecting PI3K/AKT/mTOR pathway? Is it just because as they mentioned 'Additionally, AV-IV has demonstrated its ability to safeguard against lung injury caused by PM2.5 by activating the PI3K/AKT/mTOR signaling pathway'?

They need to prove the involvement of these pathways further by using some inhibitor (of these pathways) study in vitro.

Q3. What about the toxicity if any of usage of AS-IV?

Please submit your revised manuscript by 31st March, 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: Yes

Reviewer #2: Partly

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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

**********

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: Dear Authors,

Your manuscript titled "The impact of Astragaloside IV on the inflammatory response and Gut microbiota in cases of acute lung injury examined through the PI3K/AKT/mTOR pathway" presents a comprehensive study exploring the therapeutic potential of Astragaloside IV (AS-IV) in the context of acute lung injury (ALI) and its effect on gut microbiota through modulation of the PI3K/AKT/mTOR pathway. The study is well-structured, and the methodology is robust, covering a wide range of techniques from histological analysis to gene expression and microbiota composition analysis. However, there are several areas where improvements could enhance the clarity and impact of your findings:

1. Literature Review: While your introduction provides a good background, it would be beneficial to further discuss previous studies that have explored similar treatments for ALI, particularly focusing on those involving gut microbiota modulation. This would contextualize your findings within the broader research landscape.

2. Methodological Details: In several sections, the methodology could benefit from more detailed descriptions to ensure reproducibility. For example, specifics regarding the conditions under which the 16S rRNA sequencing was performed (such as sequencing depth and bioinformatics tools used) would be valuable.

3. Statistical Analysis: While you have employed appropriate statistical tests, the manuscript would benefit from a more detailed explanation of the choice of these tests and any assumptions made. Additionally, include information on the corrections applied for multiple comparisons, if any.

4. Results Interpretation: The results are compelling but could be further strengthened by a more detailed discussion of the implications of your findings, particularly the mechanistic insights into how AS-IV modulates the gut-lung axis. Consider discussing potential limitations of the PI3K/AKT/mTOR pathway's role in mediating these effects.

5. Figures and Tables: Ensure that all figures and tables are clearly labeled and referenced in the text. Some figures, such as the graphical abstract and histological images, could benefit from higher resolution or more detailed annotations to aid interpretation.

6. Conclusion and Future Directions: The conclusion succinctly summarizes the findings but could be expanded to more explicitly state the potential clinical implications of your research and suggest specific areas for future studies.

7. References: Check the currentness of your references, especially focusing on recent advances that might complement your discussion.

8. Ethical Considerations: Your ethical approval statement is in order, but please ensure that any additional ethical considerations, particularly concerning animal welfare, are comprehensively addressed.

Reviewer #2: The present manuscript entitled 'The impact of Astragaloside IV on the inflammatory response and Gut microbiota in

cases of acute lung injury is examined through the utilization of the PI3K/AKT/mTOR

pathway.' describes efficacy of a natural triterpenoid saponin compound, Astragaloside IV(AS-IV), in the treatment of AIL by the supression of the inflammatory factors through

PI3K/AKT/mTOR pathway and optimizes the composition of the gut microflora in AIL

rats. The study has several shortcoming though is of scientific merit.

Q1. What was the rationale of selecting the dose and duration of AS-IV? What was the route of injection of AS-IV?

Q2. What is the rationale for selecting PI3K/AKT/mTOR pathway? Is it just because as they mentioned 'Additionally, AV-IV

has demonstrated its ability to safeguard against lung injury caused by PM2.5 by activating the

PI3K/AKT/mTOR signaling pathway'?

They need to prove the involvement of these pathways further by using some inhibitor (of these pathways) study in vitro.

Q3. What about the toxicity if any of usage of AS-IV?

**********

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Reviewer #1: Yes: Dr. Prasad Andhare

Reviewer #2: No

**********

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The impact of Astragaloside IV on the inflammatory response and Gut microbiota in cases of acute lung injury is examined through the utilization of the PI3K/AKT/mTOR pathway.

PONE-D-23-42843R1

Dear Dr. Jia Ke,

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.

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Kind regards,

Palash Mandal

Academic Editor

PLOS ONE