Anti-Inflammatory Activity of the Active Compounds of Sanguisorbae Radix In Macrophages and in Vivo Toxicity Evaluation in Zebrafish

Round 1

Reviewer 1 Report

This paper present the antiinflammatory properties of active compounds of Sanguisorbae Radix in macrophages and toxicity in Zebrafish

Commets:

Materials and methods

please add more data about RT-PCR reaction  pleased add data about purity of studied compounds

Results

please improve quality  immunoblots

 

Results and discussion

the authors should improve and add more example about studied compounds and structure, in my opinion about SAR

 

Author Response

Materials and methods

please add more data about RT-PCR reaction pleased add data about purity of studied compounds

 

Results

please improve quality immunoblots

Results and discussion

the authors should improve and add more example about studied compounds and structure, in my opinion about SAR

Answer: We have added ¹H NMR, ¹³C analysis data to the Sample Preparation section. We have also uploaded additional supplementary data regarding the isolation of SR.

Author Response File: Author Response.docx

Reviewer 2 Report

The authors have evaluated the anti-inflammatory effects of extracted compounds (QC, CC, and GA) using the RAW264.7 cell line and evaluated it's in vivo toxicity in zebrafish. The authors studied the molecular mechanism for the anti-inflammatory effects using ELISA. The core of this manuscript is based on the extracted compounds. However, the manuscript lacks the details information on their characterization. The authors should address the following concerns before the acceptance of the manuscript. How the extracted compounds were characterized. A complete characterization profile starting with the color of the extracted compounds, 1H NMR, 13C, and MASS spectra should be submitted in the supporting information file. The authors should also provide relevant HPLC data for each compound.

Further, the authors conclude that the SR has potential application as a safe skin anti-inflammatory biomaterial. it is understandable that they have evaluated the said activity. However, the manuscript clearly lacks the experimental data to conclude that these extracts are safe to use in skincare products as an anti-inflammatory biomaterial.

Author Response

The authors have evaluated the anti-inflammatory effects of extracted compounds (QC, CC, and GA) using the RAW264.7 cell line and evaluated it's in vivo toxicity in zebrafish. The authors studied the molecular mechanism for the anti-inflammatory effects using ELISA. The core of this manuscript is based on the extracted compounds. However, the manuscript lacks the details information on their characterization. The authors should address the following concerns before the acceptance of the manuscript. How the extracted compounds were characterized. A complete characterization profile starting with the color of the extracted compounds, 1H NMR, 13C, and MASS spectra should be submitted in the supporting information file. The authors should also provide relevant HPLC data for each compound.

Answer: I have added ¹H NMR, ¹³C analysis data to the Sample Preparation section. We also uploaded additional supplementary data on the isolation of SR.

Further, the authors conclude that the SR has potential application as a safe skin anti-inflammatory biomaterial. it is understandable that they have evaluated the said activity. However, the manuscript clearly lacks the experimental data to conclude that these extracts are safe to use in skincare products as an anti-inflammatory biomaterial.

Answer: Thank you for your comments. In regard to your concern about the claim that SR is safe for use on skin as an anti-inflammatory biomaterial, you are justified in pointing out that we did not provide complete evidence for this in our study. Instead we are relying on literature for this claim, specifically that SR has been used as a skin anti-inflammatory compound in several studies and has been proven safe in cytotoxicity experiments (Jeong et al., 2018; Yang, Yoo, Cho, & Ma, 2016).

Author Response File: Author Response.docx

Reviewer 3 Report

In this manuscript the authors evaluated the anti-inflammatory properties of three flavonoids compounds: quercetin, catechin and gallic acid extracts by Sanquisorbae radix. The authors demonstrated that the quercetin present the highest anti-inflammatory and not present the toxicity of concentration below 25 µg/ml.

The experiments have been systematically performed and the manuscript is well organized therefore I suggest that this manuscript can be published on this journal after the correction of the following issue: the graphical abstract is missing

Based on the above comments, I would suggest publishing the manuscript after minor revision.

Author Response

In this manuscript the authors evaluated the anti-inflammatory properties of three flavonoids compounds: quercetin, catechin and gallic acid extracts by Sanquisorbae radix. The authors demonstrated that the quercetin present the highest anti-inflammatory and not present the toxicity of concentration below 25 µg/ml.

The experiments have been systematically performed and the manuscript is well organized therefore I suggest that this manuscript can be published on this journal after the correction of the following issue: the graphical abstract is missing

Answer: The graphical abstract was not submitted as it is not a mandatory formatting requirement of the journal.

Author Response File: Author Response.docx

Round 2

Reviewer 1 Report

 I accept this version

Reviewer 2 Report

The authors have answered all queries.