Quantitative Analysis and Molecular Docking Simulation of Flavonols from Eruca sativa Mill. and Their Effect on Skin Barrier Function

Round 1

Reviewer 1 Report

With regard to the capacity of antioxidants

It is recommended to conduct two additional concentration tests or eliminate the dose-dependent term, since employing the specified term after testing only two doses is inappropriate.

In regard to flavonoid analysis

As supporting evidence, include the chromatogram obtained and the R^2 value of the calibration curve for quercetin and gallic acid.

Justify the use of an 84-minute program more precisely if the final peak of interest is attained in 33.9 minutes; this program could be regarded a cleaning program.

In general, 

Revision is made to the mL unit at line 208.

Conducting an assessment of the molecular formulas of AlCl3 and NaNO2 at lines 225 and 226.

Insert a modification at line 237 by including 5×105 cells.

Kindly verify the CO2 potentialization and molecular formula listed on line 270.

Author Response

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Author Response File: Author Response.pdf

Reviewer 2 Report

This study aimed to investigate the effect of Eruca sativa 70% ethanol extract (ES) and its fractions on skin barrier function. The results showed that Ethyl acetate fraction (EEA) showed the highest total polyphenol and flavonoid content. ES and EEA acted as PPAR-α ligands. EEA significantly increased cornified envelope formation in HaCaT cells. EEA reduced nitric oxide and pro-inflammatory cytokines in lipopolysaccharide-stimulated RAW 264.7 cells. Flavonol mono-glycosides from EEA were shown to be a potent PPAR-α ligand and showed the inhibition of nitric oxide.

1.         The effect of Eruca sativa extract and the flavonoids contained in the plants on antiinflammation was reported in the literature. The advance of this study must describe clearly in the manuscript.

2.         This study used the cell viability as the marker of cornified envelope formation, however, the cell viability may not a direct marker of cornified envelope formation. A suitable marker for cornified envelope formation must determine in this study.

3.         This study stated the effect of Eruca sativa extract and the flavonoids shown, skin barrier function improvement. However, the experiment on skin barrier function was lacking. Some experiments to determine the effect and markers related to skin barrier function must do and present in this manuscript.

4.         The effects of the flavonoids contained in the Eruca sativa extract on antiinflammation and skin barrier function have to discuss in depth. In addition, the results of this study needed more discussion with other work in the literature.

5.         There were some type and grammar errors in the manuscript. Please recheck the manuscript carefully.

There were some type and grammar errors in the manuscript. Please recheck the manuscript carefully.

Author Response

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Author Response File: Author Response.pdf

Reviewer 3 Report

In this article, Park et al reported the effect of Eruca sativa (ES), ethyl acetate fraction (EEA). EEA significantly increased cornified envelope formation as a keratinocyte terminal differentiation marker in HaCaT cells. Further, it significantly reduced nitric oxide and pro-inflammatory cytokines in lipopolysaccharide-stimulated RAW 264.7 cells. The main flavonol forms detected in high amounts from EEA are mono-, and di-glycoside of each aglycone. Flavonol mono-glycosides were shown to be a potent PPAR-α ligand using molecular docking simulation and showed the inhibition of nitric oxide. They concluded that E. sativa is suitable for use in improving skin barrier function and inflammation in skin disorders, such as atopic dermatitis. I have some questions.

major concerns)

1) In vitro experiments alone are not sufficient to show whether these substances actually help in skin barrier. It is necessary to use the drug in mice and humans and observe in detail the skin condition, AD model, skin moisturizing state, loricrin and filaggrin, and skin pathological state. The data presented here can only serve as a candidate for such a study, and nothing can be said about how the drug actually affects the skin barrier. Please rework your logical structure a bit more.

minor concerns)

1) In line 95, "were were" dubbed.

2) In 5: conclusions, not 5 instead "4."

3) In line 118, "many of PAPR-activator?" You mean not PAPR but "PPAR."

Author Response

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Author Response File: Author Response.pdf

Round 2

Reviewer 2 Report

1.         The authors explained that the three flavonols and their glycosides reported in the previous paper were quantified in all organic solvent fractions, and three of the flavonol mono-glycosides in particular were identified as PPAR-α activators through molecular modeling. However, the advance of this study did not describe clearly in the revised version.

2.         The results of this work have to discuss in depth and present in the text.

3.         The type and grammar errors were revised in text in the manuscript but not the Figure Legends. Please recheck the manuscript thoroughly.

Minor editing of English language required.

Author Response

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Author Response File: Author Response.pdf

Reviewer 3 Report

No additional comments.

Author Response

No additional comments.

I didn't answer because it wasn't pointed out to me.