Inhibition of Glial Activation and Subsequent Reduction in White Matter Damage through Supplementation with a Combined Extract of Wheat Bran, Citrus Peel, and Jujube in a Rat Model of Vascular Dementia
Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsGeneral comment:
The authors conducted research to investigate whether the optimized extract of wheat bran, citrus peel and jujube (WCJ) could reduce white matter damage in a rat model of vascular dementia (VaD).
The authors state that they chose WCJ as a decoction based on its potential efficacy in reducing brain damage under ischemic conditions.
Materials and methods used included cell culture experiments, TUNEL assay to measure the DNA nicks generated in the cells as a biomarker for apoptotic cells, Western Blotting, induction of bilateral common carotid artery occlusion (BCCAO) mimicking VaD in rats and rapid staining in Luxol blue to assess white matter damage.
The results showed that WCJ treatment improved cell survival, prevented astrocytic and microglial activation, attenuated myelin damage, and improved pupillary light reflex in the rat BCCAO model.
The authors concluded that consumption of WCJ has the potential to prevent VaD by reducing white matter damage, suggesting that it could be developed as a safe herbal medicine to prevent VaD.
Strengths:
1) The article is written in a coherent and easy to understand way.
2) The tables and figures are clear and suggestive.
3) The conclusions are consistent with the evidence and arguments presented.
4) References are relevant.
Minor corrections:
1) Rows 46, after occluded you must put a point.
Recommendations:
1) It is recommended to prepare a graphic abstract for a better understanding of the text
2) It is suggested to improve the quality of figures 2, 3 and 5
Author Response
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Author Response File: 
Author Response.pdf
Reviewer 2 Report
Comments and Suggestions for AuthorsThe paper is based on cel culture and animal model studies. Authords aimed to observe impact pf wheat bran, citrus peel and jujube extratc on reduction of white matter damage. The issue is very interesting and important but some points have to be improved:
Abstract: informative, includes the most important information
Introduction: based on up to date references, includes significant information about presented issue
Methods: adequate for aim of studies. Authors decided to use various analysis based on cel culture as well as in vivo models. My question is: why authors decided to use only 6 animal in each group? The numer of animals is small. Statistical analysis can be disturbed as well as reduction of the group resulted from death is highly probable. Additionally, authors decided to use only these 3 groups of animals or the study description is presented not exactly?
Results: obtained results are presented in form of figures. The statistical significance could be better but can be accepted in present form. Obtained results are interesting and significant for future consideration of similar studies.
Discussion: well prepared but the section can be enriched with more information about similar studies performed within last few years.
Conclusions: Insufficient. Improvement needed.
Author Response
Please see the attachment.
Author Response File: Author Response.pdf
Reviewer 3 Report
Comments and Suggestions for AuthorsThe manuscript reports the protective effects of a combined extract of wheat bran, citrus peel, and sliced jujube in C6 cells under hypoxic conditions and in a rat model of bilateral common carotid artery occlusion (BCCAO), which mimics vascular dementia. The findings are noteworthy; however, they highlight crucial concerns that need to be addressed.
General comments
The title is partially in accordance with the content of the manuscript. In particular, they should improve the first part of the title, “Reduction of white matter damage …” considering that they also recorded other neuroprotective effects, for example, the inhibition of glial activation.
The abstract should be improved because the combined experimental approach of an in vitro model of hypoxia and an in vitro model of vascular dementia is not clear.
Specific comments
The authors evaluated the viability of C6 cells under hypoxic conditions with trypan blue dye. They never report hypoxia-induced cytotoxicity or a decline in cell viability. In this regard, the graphic of Figure 2 is not explanatory of the results obtained with the different combinations of concentrations of Citrus and Jujube peel extracts. They should change this graphic into more graphics that are appreciated to evaluate the effects of the different combinations on cytotoxicity (cell death) or a decline in cell viability. They should also add statistical analysis.
The cell viability reported in Figure 3 is misleading. There is no cell viability greater than 100%. In Figure 3 the sham is missing.
The author reported “Grade” on the ordinate of Figure 5i. What does “Grade” mean? They should explain the grade scale used in the method section and improve the title of the ordinate.
They evaluated the C6 cell apoptosis under hypoxic conditions. Did they find apoptotic cells in the corpus callosum and optic tract? This information could also support the potential neuroprotective effects of the studied extract combination.
Comments on the Quality of English LanguageThe formation of English sentences is below the minimal expectation level for scientific journals. I recommend a review of the entire manuscript.
Author Response
Please see the attachment.
Author Response File: Author Response.pdf
Round 2
Reviewer 3 Report
Comments and Suggestions for AuthorsThe revised paper is suitable for publication
