Samnamul (Shoots of Aruncus dioicus) Inhibit Adipogenesis by Downregulating Adipocyte-Specific Transcription Factors in 3T3-L1 Adipocytes

Round 1

Reviewer 1 Report

Comment  to  the  review of Manuscript ID: processes-996473

Title: Samnamul (shoots of Aruncus dioicus) inhibit adipogenesis by
downregulating adipocyte-specific transcription factors in 3T3-L1 adipocytes.

Author should insert to their manuscript the changes as indicated below.

-Introduction Section

In the introduction or discussion  sections there is no information on the antioxidant properties of extract from  Aruncus dioicus  which was of earlier studied [9]. Why the same variety was  tested in the present work in terms of free radical scavenger.

-Materials and Methods Section.

2.1 L 73.  Authors should specify the type of lyophilizer.

2.9  L 147.   Why was an extract  the Allium spp. used to determine antioxidant activity with DPPH method and not Aruncus dioicus ? 

 2.11  L 178.    Phytochemicals ADE from A. dioicus were investigated using HPLC mode to identify and quantify principal compounds as chlorogenic, caffeic, ferulic, veratric and cinnamic acids, and flavonol –quercetin. Authors should include some information on  method have been used for identification and quantification these compounds.

References

21. L 383 . It is - Food Anal.l Methods  and should  be - Food Anal. Methods.

     

    
    

     

Author Response

Dear. Reviewer 1.

I wish to submit a research article, titled “Samnamul (shoots of Aruncus dioicus) inhibit adipogenesis by downregulating adipocyte-specific transcription factors in 3T3-L1 adipocytes” for consideration for publication in Processes.

 

- Introduction Section

1. In the introduction or discussion sections there is no information on the antioxidant properties of extract from Aruncus dioicus which was of earlier studied [9]. Why the same variety was tested in the present work in terms of free radical scavenger.

→ The sample used in the reference in [9] is a sample collected from 'Ulleung-gun, Gyeongsangbuk-do' and 'hot air-dried', but the sample used in our paper is a 'freeze-dried' sample from 'Pyeongchang, Gangwon', which is somewhat different. Therefore, we tested free radical scavenging assay.

- Materials and Methods Section.

2.1 L 73. Authors should specify the type of lyophilizer.

→ We added type of lyophilizer. “PVTFD50R, ilShinBioBase, Korea.”

2.9  L 147. Why was an extract the Allium spp. used to determine antioxidant activity with DPPH method and not Aruncus dioicus ?

→ It had modified to “…The A. dioicus extracts (40 μL) were allowed to react with 160 μL of the DPPH solution for 1 h at room temperature in the dark. Then, the absorbance was taken at 515 nm...”

 2.11  L 178. Phytochemicals ADE from A. dioicus were investigated using HPLC mode to identify and quantify principal compounds as chlorogenic, caffeic, ferulic, veratric and cinnamic acids, and flavonol –quercetin. Authors should include some information on  method have been used for identification and quantification these compounds.

→ We were performed as described previously [23]. “The elution program was as follows (B %): 8–10%, 2 min; 10–30%, 27 min; 30–90%, 50 min; 90–100%, 51 min; 100%, 60 min; 100–8%, 62 min; and 8%, 70 min. The injection volume was 10 μL, the flow rate was 1.0 mL/min, and phenols were detected at 280nm using UV light.”

- References

L 383 . It is - Food Anal.l Methods and should be - Food Anal. Methods

→ It was modified as you requested.

Author Response File: Author Response.docx

Reviewer 2 Report

Comments to the Author
The paper provides interesting data about the anti-adipogenic and anti-lipogenic effects of ADE in murine pre-adipocytes 3T3-L1 and describes the in vitro antioxidant activities of this extract, attributed to its high content in phenolic compounds.

Experiments are well-designed and results are clearly explained. However, there are some points to be considered before publication:

1. Line 21 of abstract seems to have missed some information
2. Line 111 of M&M is incomplete
3. Line 207 of results should indicate: “All ADE concentrations from 25 ug/mL inhibited TG accumulation”.
4. Line 288: This work has analysed the levels of mRNA expression of Adipoq and Ap2 genes, so names should be written in italics and lowercase
5. Line 289 indicates that the protein ACC1 has been analysed, but this result has not been shown.
6. Line 289: This sentence should indicate that the analyses has been performed in murine 3T3-L1 adipocytes.
7. Line 319 of conclusions: Authors should indicate that this study has been performed in vitro. Thus, a suggestion would be to replace the “anti-obesity activity” by another sentence more appropriate: “ADE inhibits adipogenesis and lipogenesis processes in 3T3-L1 cells by suppressing lipid droplets, TG, …. “
8. Discussion comment 1: Authors should discuss more the different effect observed in total lipid content, TG content and cholesterol content observed after treatment with ADE in 3T3-L1.
9. Overall, this reviewer feels that the results can be discussed in greater depth. The high content in ADE of phenolics such as quercetin is very striking. The anti-adipogenic activity of this and other compounds such as hesperidin, naringenin and phenolic acids such as chlorogenic, ferulic, and protocatechic acids have also been described, through their activity against the proteins and genes analysed by the authors and other described mechanisms. These results can be discussed in greater depth using bibliographic data.
10. English must be revised in deep as there are numerous errors along the manuscript, including abstract, introduction, discussion and conclusions.

Author Response

Dear. Reviewer 2.

I wish to submit a research article, titled “Samnamul (shoots of Aruncus dioicus) inhibit adipogenesis by downregulating adipocyte-specific transcription factors in 3T3-L1 adipocytes” for consideration for publication in Processes.

 

Experiments are well-designed and results are clearly explained. However, there are some points to be considered before publication:

 

1. Line 21 of abstract seems to have missed some information
   → We completed line 21.
2. Line 111 of M&M is incomplete
   → We added “TG contents of the cell lysates were measured at 570 nm using a Synergy H1 multi-plate reader” in line 111.
3. Line 207 of results should indicate: “All ADE concentrations from 25 ug/mL inhibited TG accumulation”.
   → We modified as your request in line 209.
4. Line 288: This work has analysed the levels of mRNA expression of Adipoq and Ap2 genes, so names should be written in italics and lowercase
   → The names of mRNA are modified in italics and lowercase in overall paper.
5. Line 289 indicates that the protein ACC1 has been analysed, but this result has not been shown.
   → We deleted ‘ACC1’ in line 292.
6. Line 289: This sentence should indicate that the analyses has been performed in murine 3T3-L1 adipocytes.
   → We added “…lipogenic proteins and mRNA, including SREBP1c, and FAS in 3T3-L1 adipocyte….” in line 292.
7. Line 319 of conclusions: Authors should indicate that this study has been performed in vitro. Thus, a suggestion would be to replace the “anti-obesity activity” by another sentence more appropriate: “ADE inhibits adipogenesis and lipogenesis processes in 3T3-L1 cells by suppressing lipid droplets, TG, …. “
   → We modified as your request “ADE inhibits adipogenesis and lipogenesis processes in 3T3-L1 cells by suppressing lipid droplets, TG, and cholesterol levels” in line 319.
8. Discussion comment 1: Authors should discuss more the different effect observed in total lipid content, TG content and cholesterol content observed after treatment with ADE in 3T3-L1.
   → we added discussion related lipid content, TG content and cholesterol content in line 295 and 304.
9. English must be revised in deep as there are numerous errors along the manuscript, including abstract, introduction, discussion and conclusion.
   → We had been corrected English revision.