Pharmacodynamic Evaluation and Mechanism of Ginseng Polysaccharide against Nephrotoxicity Induced by Hexavalent Chromium

Round 1

Reviewer 1 Report

In the manuscript entitled “Pharmacodynamic evaluation and mechanism of ginseng polysaccharide against nephrotoxicity induced by hexavalent chromium”, the authors employed in vivo mice model to investigate the therapeutic role of ginseng polysaccharide in kidney injury caused by hexavalent chromium. They found ginseng polysaccharide can alleviate renal injury by relieve oxidative stress and apoptosis. However, there are some issues should be addressed:

1.     Previous study has shown that ginseng has the renal protective function under the injury caused by chromium. (El-Mahalaway et al. The effect of potassium dichromate on convoluted tubules of the kidney of adult male albino rats and the possible protective role of ginseng: a histological and immunohistochemical study. The Egyptian Journal of Histology 38(2):p 157-167, June 2015)  It would be better if the authors can highlight the novelty of this study.

2.     This study is mainly focused on oxidative stress and apoptosis. However, there are some studies showed that ginseng can influence macrophage activity and the infiltration and polarization of macrophage play the essential role during kidney injury. ( Front Immunol. 2021 Aug 26;12:735014) (Cells. 2021 Nov 6;10(11):3057) (Br J Pharmacol. 2023 Apr 19. doi: 10.1111/bph.16096.) (J Cell Physiol. 2022 Jan;237(1):983-991).  It will enrich the content of this study by discussing this part by referencing these current studies.

3.     The kidney injury score can be determined from the staining and evaluate the injury level.

4.     Scale bar should be added in the staining figures.

Author Response

Response to Reviewer Comments

Dear reviewer:

We are truly grateful for your critical comments and thoughtful suggestions. Based on these comments and suggestions, we have made careful modifications to the original manuscript. Changes, additions, and deletions to the manuscript are red font. We hope the revised manuscript will get your approval. Below you will find our point-by-point responses to the comments/questions. Reviewers' comments are in black font and our responses are in red font. If you have any comments/questions, please contact us, we’ll make serious changes. Thanks again.

Major comments

Point 1: Previous study has shown that ginseng has the renal protective function under the injury caused by chromium. (El-Mahalaway et al. The effect of potassium dichromate on convoluted tubules of the kidney of adult male albino rats and the possible protective role of ginseng: a histological and immunohistochemical study. The Egyptian Journal of Histology 38(2): p 157-167, June 2015). It would be better if the authors can highlight the novelty of this study.

Response 1: Thank you very much for reviewing and commenting on this part of the manuscript. We reviewed this literature and carefully modified this section. The previous experimental results are added, and the novelty of this experiment is described again. We hope the revised content meets with your approval. If you have any other questions, please contact us, and we will revise it carefully again.

Point 2: This study is mainly focused on oxidative stress and apoptosis. However, there are some studies showed that ginseng can influence macrophage activity and the infiltration and polarization of macrophage play the essential role during kidney injury. (Front Immunol. 2021 Aug 26; 12:735014) (Cells. 2021 Nov 6;10(11):3057) (Br J Pharmacol. 2023 Apr 19. Doi: 10.1111/bph.16096.) (J Cell Physiol. 2022 Jan;237(1):983-991). It will enrich the content of this study by discussing this part by referencing these current studies.

Response 2: Thanks for your valuable suggestion. We carefully read the literature you recommended and added it to the discussion section. The mechanism of ginseng polysaccharide in alleviating nephrotoxicity induced by hexavalent chromium was discussed in a deeper level. If you have any other questions, please contact us, and we will revise it carefully again.

Point 3: The kidney injury score can be determined from the staining and evaluate the injury level.

Response 3: Thank you very much for your valuable suggestions. Based on your suggestions, we added a description to determine the degree of kidney damage from staining. We hope the revised content meets with your approval. If you have any other questions, please contact us, and we will revise it carefully again.

Point 4: Scale bar should be added in the staining figures.

Response 4: Thank you very much for your valuable suggestions. Based on your suggestions, we have carefully revised this information. The scale bars were added to the stained graph. We hope the revised content meets with your approval. If you have any other questions, please contact us, and we will revise it carefully again.

We believe that these changes have addressed the issues raised by the reviewer, and have significantly improved the clarity and impact of our work.

Once again, we would like to express our sincere gratitude to you for your expert review and thoughtful comments. Your feedback has been instrumental in strengthening our research, and we are grateful for the opportunity to benefit from your expertise.

Please do not hesitate to let us know if you require any further information or clarification. We look forward to hearing back from you soon.

Sincerely,

Xin Zhou

Author Response File: Author Response.docx

Reviewer 2 Report

This study investigated the protective effects and mechanisms of ginseng polysaccharides against hexavalent chromium-induced renal toxicity. Further, it employed lipidomic correlation analysis to explore how ginseng polysaccharides protect the organism by modulating the expression of various lipids. While this research held significant meaning, there are several issues to address:

1. The mechanism underlying hexavalent chromium-induced renal toxicity in mice was not comprehensively delineated in the background section.

2. In the introduction, the study provided relatively limited background information and insufficient research on ginseng polysaccharides specifically for their pharmacodynamic effects against hexavalent chromium's renal toxicity.

3. The presentation of figures and legends is unclear. It is essential to explicitly state the subject populations and experimental conditions for each study. For instance, doses of MC, PC, and GPS1-4 are not distinctly marked in Figure 4. Similarly, distinctions between Tables 4 and 5, Figures 1 and 2, and Figures 4 and 5 are not clearly articulated. Corresponding descriptions in the results section can lead to misinterpretation, such as lines 259-263 and lines 265-271.

4. Statistical descriptions are inadequately detailed; while ANOVA is used for multiple comparisons, the method employed for pairwise comparisons is not specified.

5. The authors do not provide a description of the rationale behind the selection of hexavalent chromium doses.

6. There are problems with some of the authors' descriptions of results, which lack rigor. For example, line 256 states "The differences at different doses are different," which is vague. Also, on lines 248-249, it says, "However, compared to the MC group, the impacts of the PC and GPS groups were much lower." It's unclear how 'much lower' was determined. Such instances abound throughout the text, requiring the author to thoroughly revise these sections.

7. There are substantial language issues within the manuscript, and it is recommended that the authors engage a professional person for a comprehensive revision.

There are substantial language issues within the manuscript, and it is recommended that the authors engage a professional person for a comprehensive revision.

Author Response

Response to Reviewer Comments

Dear reviewer:

We are truly grateful for your thoughtful comments. Based on these comments and suggestions, we have made careful modifications to the original manuscript. Changes, additions, and deletions to the manuscript are red font. We hope the revised manuscript will get your approval. Below you will find our point-by-point responses to the comments. Reviewers' comments are in black font and our responses are in red font. If you have any comments/questions, please contact us, we’ll make serious changes. Thanks again.

Point 1: The mechanism underlying hexavalent chromium-induced renal toxicity in mice was not comprehensively delineated in the background section.

Response 1: Thank you for your professional comment. We think these are excellent suggestions. Based on your comments, In the introduction, we add a description of the mechanism of hexavalent chromium-induced nephrotoxicity in mice. We hope to gain your approval. If you have any other questions, please contact us, and we will revise it carefully again.

Point 2: In the introduction, the study provided relatively limited background information and insufficient research on ginseng polysaccharides specifically for their pharmacodynamic effects against hexavalent chromium's renal toxicity.

Response 2: Thank you for your professional comment. Following your suggestion, we added the content of pharmacodynamics research background in the introduction and added references to support it. We hope to gain your approval. If you have any other questions, please contact us, and we will revise it carefully again.

Point 3: The presentation of figures and legends is unclear. It is essential to explicitly state the subject populations and experimental conditions for each study. For instance, doses of MC, PC, and GPS1-4 are not distinctly marked in Figure 4. Similarly, distinctions between Tables 4 and 5, Figures 1 and 2, and Figures 4 and 5 are not clearly articulated. Corresponding descriptions in the results section can lead to misinterpretation, such as lines 259-263 and lines 265-271.

Response 3: Thank you very much for this useful information. We have rewritten this paragraph. In our notes, we checked all the charts and added the corresponding notes, and redescribed them in the results section, making the results of the aging experiment and the pharmacodynamic experiment more clearly expressed. We hope the revised content meets with your approval. If you have any other questions, please contact us, and we will revise it carefully again.

Point 4: Statistical descriptions are inadequately detailed; while ANOVA is used for multiple comparisons, the method employed for pairwise comparisons is not specified.

Response 4: Thank you for your careful examination and valuable suggestions. We feel sorry for our carelessness. We give a supplementary description of this part. “Differences between different groups were compared with Student’s t-test”. We hope to gain your approval. If you have any other questions, please contact us, and we will revise it carefully again.

Point 5: The authors do not provide a description of the rationale behind the selection of hexavalent chromium doses.

Response 5: Thank you very much for your valuable suggestions. We give a supplementary description of this part. And we hope the revised content meets with your approval. If you have any other questions, please contact us, and we will revise it carefully again.

Point 6: There are problems with some of the authors' descriptions of results, which lack rigor. For example, line 256 states "The differences at different doses are different," which is vague. Also, on lines 248-249, it says, "However, compared to the MC group, the impacts of the PC and GPS groups were much lower." It's unclear how 'much lower' was determined. Such instances abound throughout the text, requiring the author to thoroughly revise these sections.

Response 6: Thank you for your careful examination and valuable suggestions. We feel sorry for our carelessness. We have carefully reviewed and revised it in the manuscript. And we hope to gain your approval. If you have any other questions, please contact us, and we will revise it carefully again.

Point 7: There are substantial language issues within the manuscript, and it is recommended that the authors engage a professional person for a comprehensive revision.

Response 7: Thank you very much for reviewing and commenting on this part of the manuscript. As authors whose first language is not English, we recognize that English expression is our weakness. However, in order to be as accurate as possible, we have enlisted the help of individuals with good English proficiency to assist with language editing when revising the manuscript. We hope that the revised manuscript can meet your approval.

We believe that these changes have addressed the issues raised by the reviewer, and have significantly improved the clarity and impact of our work.

Once again, we would like to express our sincere gratitude to you for your expert review and thoughtful comments. Your feedback has been instrumental in strengthening our research, and we are grateful for the opportunity to benefit from your expertise.

Please do not hesitate to let us know if you require any further information or clarification. We look forward to hearing back from you soon.

Sincerely,

Xin Zhou

Author Response File: Author Response.docx

Round 2

Reviewer 1 Report

The authors addressed my concerns.

Author Response

Response to Reviewer Comments

Dear reviewer:

Thank you very much for taking the valuable time to review my manuscript again. You did not put forward new ideas in the second review, which is an encouragement to me, and also let me have more confidence in the quality of the manuscript.

I have carefully checked the manuscript to confirm that all previous modifications have been completed, and the manuscript is now ready to move on to the next stage. If you have any other suggestions or opinions in the future, I am still very willing to accept and implement them.

Thank you again for your hard work and professional advice.

Sincerely,

Xin Zhou

Author Response File: Author Response.docx

Reviewer 2 Report

The author has essentially addressed some of the previous comments. However, there is an issue with the statistical analysis method employed. Pairwise comparisons should not be conducted using a t-test in an ad hoc manner.

There are a few grammatical issues present, but they are generally acceptable.

Author Response

Response to Reviewer Comments

Dear reviewer:

We are truly grateful for your thoughtful comments. Based on these comments and suggestions, we have made careful modifications to the original manuscript. Changes, additions, and deletions to the manuscript are highlighted. We hope the revised manuscript will get your approval. Below you will find our point-by-point responses to the comments. Reviewers' comments are in black font and our responses are in red font. If you have any comments/questions, please contact us, we’ll make serious changes. Thanks again.

Point 1: However, there is an issue with the statistical analysis method employed. Pairwise comparisons should not be conducted using a t-test in an ad hoc manner.

Response 1: Thank you for your careful examination and valuable suggestions. We feel sorry for our carelessness. We change this part to “All the experimental data were analyzed by multiple analysis using the social science statistical package (SPSS 26 version), expressed as mean ± standard deviation (mean ± SD). One-way analysis of variance was used for inter-group differences(ANOVA), p < 0.05 indicates a significant difference, and p < 0.01 indicates a highly significant difference (compared with the blank control group: *p < 0.05, **p < 0.01; compared with the model group: ^▲▲p < 0.05, ^▲▲p < 0.01)”. We hope to gain your approval. If you have any other questions, please contact us, and we will revise it carefully again.

Point 2: There are a few grammatical issues present, but they are generally acceptable.

Response 2: We feel great thanks for your professional review word on our article. English is indeed our weakness. We have tried to make modifications again and marked the modified part as highlighted. We hope to gain your approval. If you have any other questions, please contact us, and we will revise it carefully again.

We believe that these changes have addressed the issues raised by the reviewer, and have significantly improved the clarity and impact of our work.

Once again, we would like to express our sincere gratitude to you for your expert review and thoughtful comments. Your feedback has been instrumental in strengthening our research, and we are grateful for the opportunity to benefit from your expertise.

Please do not hesitate to let us know if you require any further information or clarification. We look forward to hearing back from you soon.

Sincerely,

Xin Zhou

Author Response File: Author Response.docx