Comparison of Steviol Glycosides or Stevia Leaves Addition to the Hypercholesterolemic Diet on Selected Biochemical Parameters of Experimental Rats

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

No comments for the authors.

Author Response

Dear Reviewer

The authors thank you very much for the review. We tried to improve the manuscript and hope that the corrections will be sufficient and certainly will get better this paper.

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

The present research compared the effects of dietary steviol glycosides and Stevia leaves addition on the biochemical parameters of rats fed on hypercholesterolemic diet, and has important theoretical and applied significance. However, the manuscript has some weaknesses and need to be improved.

1.     There are language and technical mistakes in the presentation of the manuscript. Author should check and improve the manuscript.

2.     Some acronyms and abbreviations had no definition or explanation on its first mention in the text. Please check and add relevant content.

3.     In Materials and Methods, the sections of live were stained with PAS to assess the glycogen content, but I can’t find the corresponding results in Result section.

4.     The description of some results is too verbose.

5.     I can’t find the blue arrow in figure 1, but the note of figure 1 explained the blue arrow.

6.     The discussion is more focused on the enumeration of similar studies. Analysis of the reasons behind the results is lacking.

7.     Line 416-417: In our studies, it was found that the addition of steviol glycosides to a hypercholesterol diet caused a significant reduction in serum triacylglycerols. But Table 2 showed that the triacylglycerols content had no significant difference between CHOL and GS groups. Please check this sentences and Table 2.

8.     Table 2 indicated that the activities of AST and ALT in GS group were low than those in CHOL group, but the difference is not significant. In Discussion and Conclusion, author considered that GS diet could decrease the ALT and AST activities. Author should draw more accurate conclusion and discussed precisely.

9.     Experimental diets had no significant effects on the activities of SOD, GR and GSH-Px, author can discuss these parameters together.

10.  The morphology of liver is not discussed, author only descripted the morphology results again in Discussion.

11.  In Conclusion, the content is too redundant.

Author Response

Dear Reviewer

 

The authors thank you very much for a accurate and very factual review. We tried to improve the manuscript in accordance with the comments and remarks, which certainly get better this paper.

 

Answers for Comments and Suggestions of Reviewer 2

 

 

The review form 2

 

The present research compared the effects of dietary steviol glycosides and Stevia leaves addition on the biochemical parameters of rats fed on hypercholesterolemic diet, and has important theoretical and applied significance. However, the manuscript has some weaknesses and need to be improved.

 

1. There are language and technical mistakes in the presentation of the manuscript. Author should check and improve the manuscript.

The manuscript has been corrected and lenguage has been improved. The authors hope that the posted corrections will be sufficient.

 

2. Some acronyms and abbreviations had no definition or explanation on its first mention in the text. Please check and add relevant content.

 

The authors checked and corrected the manuscript according to the review’s recommendations.

 

3. In Materials and Methods, the sections of live were stained with PAS to assess the glycogen content, but I can’t find the corresponding results in Result section.

The authors removed the following part of the sentence „and PAS (Periodic acid-Schiff) for the presence of glycogen” – lines 164-165

4. The description of some results is too verbose.

Can we please the reviewer for agreeing to leave the text unchanged. The authors consider a more detailed description of the obtained results not as inappropriate but as suitable for better transparency of the results.

 

5. I can’t find the blue arrow in figure 1, but the note of figure 1 explained the blue arrow.

We apologize for the technical error. When pasting, the same photo was duplicated twice. However, the photo with the blue arrow has been omitted. This error has been corrected by replacing the photo with the correct one.

 

6. The discussion is more focused on the enumeration of similar studies. Analysis of the reasons behind the results is lacking.

The following part of text was added to the end of Discussion chapter “To summarize, it is extremely difficult to conclude why the addition of whole Stevia leaves to the diet caused changes mainly in the blood lipid profile while the addition of steviol glycosides alone did not alter these parameters. As mentioned above a plant is a carrier of many nutritional and non-nutritive ingredients that interfere and can trigger different, sometimes unexpected, biological effects. In order to precisely explain the obtained results, different combinations of the individual ingredients would have to be used both together and separately in similar studies, which is not easy as long-term studies involving experimental animals must be compliant with the 3R principle.”.

 

7. Line 416-417: In our studies, it was found that the addition of steviol glycosides to a hypercholesterol diet caused a significant reduction in serum triacylglycerols. But Table 2 showed that the triacylglycerols content had no significant difference between CHOL and GS groups. Please check this sentences and Table 2.

Authors thank the reviewer for valuable opinion. Authors checked the results in the table 2 and changed the sentence in line 428-430.

 

8. Table 2 indicated that the activities of AST and ALT in GS group were low than those in CHOL group, but the difference is not significant. In Discussion and Conclusion, author considered that GS diet could decrease the ALT and AST activities. Author should draw more accurate conclusion and discussed precisely.

Authors thank the reviewer for valuable opinion. Authors checked the results in the table 2 and changed the sentences in line 496-505.

 

9. Experimental diets had no significant effects on the activities of SOD, GR and GSH-Px, author can discuss these parameters together.

The authors believe that due to the ambiguous results of different authors, it is more advantageous to quote them and discuss separately. From this reason authors ask the reviewer to stay this part of discussion unchanged.

10. The morphology of liver is not discussed, author only descripted the morphology results again in Discussion.

Authors would like to emphasise that there are no research results in the available literature regarding the effect of steviol glycosides and stevia leaves on histological changes in the liver of healthy rats fed the hypercholesterolemic diet.

11. In Conclusion, the content is too redundant.

The authors are very sorry, but other reviewers expected more detailed information, so the conclusion has been enriched with their suggestions.

Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

In the manuscript, the authors  compared the effect of adding pure steviol glycosides or Stevia leaves to the hypercholesterolemic diet on selected growth and biochemical parameters of experimental (non-diabetes) rats. The results of this work contained some interesting data. However, the manuscript needs revision and some issues still need to be improved.

- There are no significant differences on selected growth and biochemical parameters between the animals fed on addition of pure steviol glycosides or Stevia leaves to the hypercholesterolemic diet  and animals fed on the hypercholesterolemic diet. If the author can prolong the experiment time？

-After 6 weeks feeding experiment，is there any disease or health risk on the animals fed on the hypercholesterolemic diet?

-line554， “resulte“ should be revised to “result“.

-The innovation and significance of the research work are not outstanding or have not been well refined. Is there any relationship between the hypercholesterolemic diet and diabetes?

Author Response

Dear Reviewer

 

The authors thank you very much for the review. We tried to improve the manuscript and hope that the corrections will be sufficient and certainly will get better this paper.

 

In the manuscript, the authors  compared the effect of adding pure steviol glycosides or Stevia leaves to the hypercholesterolemic diet on selected growth and biochemical parameters of experimental (non-diabetes) rats. The results of this work contained some interesting data. However, the manuscript needs revision and some issues still need to be improved.

- There are no significant differences on selected growth and biochemical parameters between the animals fed on addition of pure steviol glycosides or Stevia leaves to the hypercholesterolemic diet  and animals fed on the hypercholesterolemic diet. If the author can prolong the experiment time？

-After 6 weeks feeding experiment，is there any disease or health risk on the animals fed on the hypercholesterolemic diet?

Due to the limitations of the Bioethics Committee and the 3R principle, the authors were obliged to maintain appropriate experimental conditions. The hypercholesterolemic diet was used to induce oxidative stress and change the lipid profile, as in many similar studies carried out over a similar period of time. This period of time is sufficient to obtained the expected results, which is confirmed by numerous studies.

 

-line554， “resulte“ should be revised to “result“.

It was corrected.

-The innovation and significance of the research work are not outstanding or have not been well refined. Is there any relationship between the hypercholesterolemic diet and diabetes?

The authors' intention was to induce oxidative stress and changes in the lipid profile by using the hypercholesterolemic diet according to the procedure which is presented in many other studies.

The authors improved lines 59-65 according to the reviewer suggestion.

Author Response File: Author Response.pdf

Reviewer 4 Report

Comments and Suggestions for Authors

1)Line 56. “It must to be emphasized that most of the available studies connected with steviol 59 glycosides and Stevia leaves were carried out using diabetic rats,” the related references should be given at the end of the sentence

2)The needs to study the effect of stevia leaves on healty rats should be explained. The disadvantages of studying those effects with diabetic rats should be explained in a good manner to justify the selection of the subject

3) Line 106. Please revise the sentence. It is not clear what is meant in the sentences.

4) The selection of the amount of püre Stevia and its leaves should be explained with the light of the studies reported in the literature

5) The reason why different origin of stevia was used should be explained,

Comments on the Quality of English Language

Fine

Author Response

Dear Reviewer

The authors thank you very much for the review. We tried to improve the manuscript and hope that the corrections will be sufficient and certainly will get better this paper.

Answers

1)Line 56. “It must to be emphasized that most of the available studies connected with steviol 59 glycosides and Stevia leaves were carried out using diabetic rats,” the related references should be given at the end of the sentence

2)The needs to study the effect of stevia leaves on healty rats should be explained. The disadvantages of studying those effects with diabetic rats should be explained in a good manner to justify the selection of the subject.

The authors’ intention was to show for the first time the influence the separated Stevia glucosides or Stevia leaves on the selecters parameters of rats fed the hypercholesterolemic diet that should cause an oxidative stress and changes in the lipid profile. Numerous publications use this procedure to achieve this goal.

3) Line 106. Please revise the sentence. It is not clear what is meant in the sentences.

The authors wanted to explain that the addition of 10, 17, 18% addition of different origin Stevia leaves to the  hypercholesterolemic diet contained 3.36 mg of steviol glycosides/kg b.w./day in each case (as in GS group), what was determined by analyzing the content of these compounds in the plant by liquid chromatography.

4) The selection of the amount of püre Stevia and its leaves should be explained with the light of the studies reported in the literature

The publication no 3 provides the content of steviol glycosides in the leaves used in our studies, and publication 23-24 provides the amounts of steviol glycosides used in similar animals studies. This information was presented in line 111.

 

5) The reason why different origin of stevia was used should be explained,

Authors used three different Stevia leaves because the results of analyzes of the composition of these stevia leaves, which were then used for animal studies, differed, as shown in publication no 3.

In the first part of Discussion chapter the author wrote “Different climatic, soil and agrotechnical conditions affect the differences in composition of plants i.e. Stevia leaves. Therefore, it was assumed that the sum of plant components that interfere with each other weakening or strengthening the response will have an unequal effect on the evaluated parameters”.

Author Response File: Author Response.pdf

Reviewer 5 Report

Comments and Suggestions for Authors

The current manuscript is not suitable for publication due to several gaps identified during the review process. Additionally, the data presented is not adequate for an original article. There are instances where references are not directly related to the corresponding text in the manuscript. It appears that the manuscript may have been intended for submission to a different, more relevant journal.

Comments on the Quality of English Language

The manuscript needs to be reviewed by a native speaker of the language

Author Response

Dear Reviewer

The authors thank you very much for the review. We tried to improve the manuscript and hope that the corrections will be sufficient and certainly will get better this paper.

Round 2

Reviewer 2 Report

Comments and Suggestions for Authors

The revised paper manuscript has shown some improvement, but there are still several details that need to be addressed. For instance, in line 2, "weighted" should be corrected to "weighed." Additionally, some abbreviations used in the tables should be consistent with the descriptions in the paper. The authors should thoroughly review the paper and make necessary revisions to enhance its quality.

Author Response

Response to Reviewer  Comments

 

Dear Reviewer,

We would like to kindly thank you for the insightful review of our manuscript. We appreciate that you decided to accept our manuscript after all corrections.

 

The revised paper manuscript has shown some improvement, but there are still several details that need to be addressed. For instance, in line 2, "weighted" should be corrected to "weighed." Additionally, some abbreviations used in the tables should be consistent with the descriptions in the paper. The authors should thoroughly review the paper and make necessary revisions to enhance its quality.

The authors have corrected the manuscript according to reviewer suggestions.

The authors have standardized the descriptions in the tables and in the text of manuscript.

 

In behalf of co-authors, once again thank you for your valuable comments.

 

Ewa Piątkowska

Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

The authors have made the necessary modifications, therefore the manuscript can be published in this revised form.

Author Response

Response to Reviewer  Comments

 

Dear Reviewer,

We would like to kindly thank you for the insightful review of our manuscript. We appreciate that you decided to accept our manuscript after all corrections.

In behalf of co-authors, once again thank you for your valuable comments.

 

Ewa Piątkowska

Author Response File: Author Response.docx

Reviewer 5 Report

Comments and Suggestions for Authors

I rejected the current manuscript at the first review. But, with respect to the other reviewers I accept it. 

Comments on the Quality of English Language

Dear Authors, 

 

I initially rejected the manuscript during the first review. However, considering the feedback from the other reviewers, I have decided to accept it. I appreciate the input and perspectives provided by the reviewers, and I believe their insights have contributed to the improvement of the manuscript. Thank you for considering my decision.

 

Good luck

Author Response

Response to Reviewer  Comments

 

Dear Reviewer,

We would like to kindly thank you for the insightful review of our manuscript. We appreciate that you decided to accept our manuscript after all corrections.

In behalf of co-authors, once again thank you for your valuable comments.

 

Ewa Piątkowska

Author Response File: Author Response.docx