Comprehensive Dose–Response Analysis of the Effect of Ionizing Radiation on Hepatic Enzyme Parameters in a Rabbit Model

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

This manuscript investigates the effect of varying radiation doses on rabbit liver function by measuring AST, ALT, and ASP enzyme levels. The study addresses a relevant topic and is built on a clear, straightforward experimental design.

However, the data presentation and interpretation require significant improvement. In its current form, the manuscript lacks critical methodological details and sufficient context for the results, which prevents the data from robustly supporting the conclusions. Addressing the following major points is needed:

1. Describe how rabbits were distributed across groups to ensure weight (3-5 kg) was balanced and did not act as a confounding variable. The rationale for selecting the specific radiation doses (e.g., 0.053 Gy, 0.84 Gy) needs to be clearly explained.
2. Critical reagents and equipment must be specified. Please list the manufacturer and model for the biochemistry analyzer, associated software, and the centrifuge (and clarify the speed) used for serum preparation.
3. The results would be significantly strengthened by including reference ranges for AST and ALT in healthy rabbits. Furthermore, providing known values for these enzymes in rabbits with non-radiation-induced hepatitis (e.g., toxin-induced) would greatly help to better assess their biological significance.
4. Figure 1 needs to clarify x and y axis, and further explain the difference seen at time point 1. Table 1 needs to clarify the animal number under each group and add further details. Apply the same clarification for Table 2 as well as ALT and ASP sections
5. Table 3 missed data for weeks beyond week1.

While the study has potential, the current manuscript requires major revision. The conclusions are not fully supported by the data due to insufficient methodological detail, and unclear presentation of figures and tables.

Comments on the Quality of English Language

The English is acceptable.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

This study aimed to characterize the effect of radiation on liver damage using blood AST, ALT, and ALP levels as indicators. The experimental design  is straightforward . However, the results are poorly presented and difficult to follow. Below are some suggestions.

1. Please change the time labels in all figures to day 0, day1, day3, day5... This would give clear meanings to the x-axis label.
2. The samples are collected on day 0, post-radiation day 1, 3, 5, 7, 14, 21, 28. It is unclear why in the tables day 2 is shown, since no samples are collected on day 2. This is particularly confusing.
3. In Table 1&3, it is not clear the data were based on which radiation dosage group. 
4. Please explain how the group average values were obtained in Table 2&4. Which time points are used?
5. Why ALT data in Table 3 have only 5 time points?

Comments on the Quality of English Language

Some sentences (line 177 for example) should be corrected.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

The study explores the relationship between ionizing radiation and hepatic injury by measuring serum enzyme markers (AST, ALT, ALP) in rabbits. The study is relevant, since radiation-induced organ toxicity is an ongoing concern in radiotherapy, occupational exposure, and accidental exposure scenarios. The use of a dose-response framework and repeated follow-up adds rigor and allows the detection of both acute and delayed effects.

However, the manuscript in its current form requires substantial refinement. The rationale is sound, but the presentation is heavy and somewhat repetitive, the interpretation of results is sometimes overstated, and important methodological details are missing. Clarifying the experimental setup, tightening the language, and situating the findings in the broader context of radiation biology would make the paper more impactful.

Strengths

1. Relevance of Topic
   Radiation-induced organ injury is highly relevant in medical, occupational, and environmental contexts. Focusing on liver enzyme changes is clinically meaningful.
2. Experimental Design

             Use of multiple radiation doses (0.053–0.84 Gy) allows dose–response relationships to be assessed; Multiple timepoints (days to weeks) capture both acute and sub-acute changes.

1. Statistical Analysis
   Application of Generalized Linear Mixed Models (GLMM) is suitable for repeated measures and adds robustness to the findings.
2. Findings

Demonstrates significant radiation-induced alterations in AST, ALT, and ALP; Clear evidence of dose- and time-dependent effects; Results are connected to oxidative stress and free radical biology, a biologically plausible mechanism.

 

Weaknesses & Areas for Improvement

1. Clarity of Writing

The manuscript is verbose, with frequent repetition of results already presented in tables.

Sentences are long and sometimes difficult to follow.

Grammatical errors (e.g., “may be hazardous to living beings including human”) need correction.

1. Organization of Results

Tables and post-hoc comparisons are overwhelming. Summarizing key findings in text would improve readability.

Figures are mentioned but not adequately described. Ensure clarity in figure legends.

1. Interpretation Issues

ALT results are inconsistent (decrease initially, increase later; no group significance). This needs clearer explanation.

Over-interpretation in places: authors attribute enzyme fluctuations directly to oxidative stress or cell membrane changes without direct biochemical evidence.

No histological confirmation of liver injury, which would strengthen conclusions.

1. Discussion Gaps

Needs more critical comparison with prior studies (e.g., why ALT results differ from other reports).

Lacks discussion of species differences and how findings translate to humans.

 

Suggestions for Improvement

1. Improve Writing & Flow

Simplify language and reduce redundancy.

1. Strengthen Results Presentation

Summarize key findings in text, delegate detailed p-values to supplementary tables.

Provide clear figure legends that guide interpretation.

1. Deepen Discussion

Critically analyze why ALT behaved differently.

Relate findings to clinical contexts (e.g., radiation therapy patients).

1. Technical & Formatting Cleanup

Correct grammar and typos.

Ensure all references are updated and relevant.

 

Conclusion

This paper has the potential to contribute meaningful insights into radiation-induced liver injury, but it requires significant revision for clarity, organization, and critical analysis. Stronger emphasis on mechanistic evidence and clinical relevance would enhance its impact.

 

Comments on the Quality of English Language

N/A

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Round 2

Reviewer 2 Report

Comments and Suggestions for Authors

The manuscript is much improved after revision. However, there are some issues. For example, the number of animals in all tables should be clearly shown. It is unclear whether N is 6 or more (if all animals of different time points or radiation dosages are used to calculate the average values) . More importantly,  author should prepare a figure or table for the raw values of each dosage group at each time point. This would give a clear view on how the animals respond to each radiation dosage at different time points. 

Author Response

Comment 1: The manuscript is much improved after revision. However, there are some issues. For example, the number of animals in all tables should be clearly shown. It is unclear whether N is 6 or more (if all animals of different time points or radiation dosages are used to calculate the average values).

Response 1: We appreciate the reviewer's observation, we agree with this comment, the animals in all tables is written clearly, N=6 meaning each groups contain 6 rabbits in both the groups and at the time points.

Comment 2: More importantly, author should prepare a figure or table for the raw values of each dosage group at each time point. This would give a clear view on how the animals respond to each radiation dosage at different time points. 

Response 2: We thank the reviewer for highlighting this we have now added the tables with raw values of each dosage group at each time point with clear description.