PONE-D-24-20060Saikosaponin A alleviates depressive-like behaviour induced by reserpine in mice by regulating gut microflora and inflammatory responsesPLOS ONE

Dear Dr. Yin,

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Academic Editor

PLOS ONE

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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: No

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: No

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: No

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: 1. The overall English writing is not very standard; the author should revise it to make the English writing more standard.

2. The authors also write carelessly, with many typos in the manuscript, which could have been easily avoided if they had double-checked the manuscript before submitting it.

3. What is the sex of the mice? Are they all male or female? The authors should indicate this in the main text or materials and methods.

4. Line 68: “hydroxytryptamine(5-HT)” should be “hydroxytryptamine (5-HT)” with a space before the parenthesis. The same applies to the entire manuscript.

5. Line 221: “statis-tically” should be “statistically.”

6. Line 236: What does “CG” mean? Does it mean “control group”? The author should write the full name when using the abbreviation for the first time. The same applies to “HE,” “FJB,” etc.

7. Figure 1: What is the meaning of letters “a,” “b,” “c,” “d”? The authors write: “Bars with different lowercase letters indicate significant difference (p < 0.05).” But which groups were compared to get the significant differences? The authors should clearly write this in the figure legends or indicate it in the figures. The same applies to all the figures.

8. Line 258: Another section “3.2”? This should be section “3.3”, as there is already a “3.2.”

9. Line 259: Grammar/typo: “Hippocampal neuronal damage and reduction as a marker of clinical depression.” Maybe the authors mean “Hippocampal neuronal damage and reduction is a marker of clinical depression.”?

10. Line 266: Typo: “RSP+SSA group This indicates that” should be “RSP+SSA group. This indicates that.”

11. For Figure 3, the authors should quantify the number of positive cells in the hippocampus before claiming that “The findings indicate a significant increase in the number of positive cells within the hippocampus of mice in the RSP group when compared to the CG group” (Line 263-265). The same applies to Figure 3B. The author should quantify the HE staining results to show that there are “reduced neuronal cells in the hippocampus with structural abnormalities.” The same applies to Figure 4.

12. Figure 4A: The staining in Figure 4A is too faint to see. The author should quantify the signal to show the differences.

Reviewer #2: With this manuscript titled “Saikosaponin A alleviates depressive-like behaviour induced by reserpine in mice by regulating gut microflora and inflammatory responses” the authors found SSA effectively ameliorates RSP-induced depressive-like symptoms in mice through several aspects involving monoaminergic transmitters, inflammation, oxidative stress and gut microbiota.

The manuscript is pleasant to read. But I still have some concerns that need to be addressed.

Comments:

1) Since the current study was only done in male mice, author should discussion the limitation of sex difference effect of SSA.

2) Page 2, line 119. How many mice were used in the study? I noted some group have N=10 mice, but other groups have N=3 mice. Total mice?

3) It’s not clear to reader that Solvent is the vehicle for RSP or not.

4) Page 7, line 122, administration of RSP is ip or po?

5) Figure 1. Groups with N=3 mice are smaller N number for the behavior assay; it will be good include p and F value for each figure from One-way ANOVA test.

6) Any acute effect of SSA on Day 1 treatment compared to 14 days SSA treatment effect? Any dose-dependent manner of SSA in low, middle and high dose?

7) Any PK profile and half-life data of SSA?

8) It’s not clear to the reader that the behavior tests were conducted at dark or light phase. As the rodent is notational animals, the phenotype of behavior will be difference between dark and light phase. Authors should consider and discuss it.

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Reviewer #1: No

Reviewer #2: No

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Saikosaponin A alleviates depressive-like behavior induced by reserpine in mice by regulating gut microflora and inflammatory responses

PONE-D-24-20060R1

Dear Dr. Yin,

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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If your institution or institutions have a press office, please notify them about your upcoming paper to help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org.

Kind regards,

Peng Zhong, Ph.D.

Academic Editor

PLOS ONE

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: All comments have been addressed

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: Yes

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

Reviewer #1: Yes

Reviewer #2: Yes

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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: Yes

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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: Yes

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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: (No Response)

Reviewer #2: The revised version of manuscript is better than previous version. For one of my questions, the author should clarify the behavior tests were done in light or dark phase as rodent is nocturnal animals.

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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

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