Augmentation and Evaluation of an Olive Oil Based Polyherbal Combination against Diabetic Cardiomyopathy in Experimental Model of Rodents

Round 1

Reviewer 1 Report

In this study, Shamim et al. evaluate the role of Olive oil-based herbal combination for diabetic cardiomyopathy. They utilized the STZ+HFD rodent model for the experimental evaluation of their herbal combination. By checking different parameters, theyy observed that the polyherbal combination shows protective effects. However, although the results are interesting, there are several major concerns that limit the enthusiasm in this study.

 

11. The major concern is the writing style of this study: This is a poorly written article with a complete lack of organization, innumerable grammatical errors, & unclear figure format and structures (sizes). Although the results might be conveying something interesting, the way it has been described makes it difficult to understand.

T2. The second major concern is lack of experimental controls. The authors have used STZ +HFD mediated beta cell dysfunction. However, both of these agents have an independent mechanism of action. The study warrants the inclusion of controls with separate STZ and HFD treatment.

33. The authors have not standardized the dosage of PHC-6 and PHC-10 based on the weight of the mice. They fed 2 mL of the PHC. The dosages need to be controlled.

 

44. The composition of the normal pellet diet is not mentioned.

55.  The figure legends are not structured and explained appropriately.

66.  Explain the food efficiency ratio.

77. The introduction only speaks about the background of different herbs and olive oil. It needs to include the missing gaps in the literature, the questions the authors had, the rationale, and the hypothesis. Similarly, the discussion needs to be more scholarly. At this stage, it looks like a mere re-iteration of the results. Finally, the methods section needs to be extensive – The authors fail to explain major methods including how the metabolites, triglycerides, enzymes, and collagen were measured in this section.

88. The jargon of the article is extremely frail: To point a few examples –

a.       “and the high-fat eating regimen taking care of was continued on for a further week to prompt a type II diabetic aggregate with changed heart digestion”

b.       “The results of D-HFD-C group were found to be very highly significantly different when compared with Normal Control group”

c.       The smudge was made by responding with hydrogen peroxide and diaminobenzidine

Author Response

Please find the reply to the Reviewer's Comments attached as a word file below.

Author Response File: Author Response.docx

Reviewer 2 Report

Shamim et al. provide a noval polyherbal combination (PHC) therapy against the diabetic cardiomyopathy using a well-established rat model. The results demonstrated PHC (PHC-6 and PHC-10) to have protective effects that were potentially beneficial compared with standard drugs metformin and carvedilol.

 

There are several critical issues:

Major comments:

1, The rationale for using the PHC-C6 and PHC-C10, instead of the other specific herbs, needs to be further highlighted. Based on all the findings, the evidence of the advantage of PHC-C6/10 compared to metformin and carvedilol is weak. The study lacks innovation and creativity.

2, The author should illustrate how the components of the three herbs: Tinospora cordifolia, Withania somnifera and Boerrhavia diffusa were mixed in PHC-C6 and C-10.

3, In the Methods section, there is an inconsistency in the application does of the STZ. In line 150, the authors indicated ‘Streptozotocin 35 mg/Kg body weight’, while in Table 1., the author listed as ‘Single i.p. injection of streptozocin at the dose of 30 mg/kg b.w.’.

4, As the author quoted that the capacity of the β-cell is essential to the pathology process of the cardiomyopathy. It is suggested that authors should perform experiments to address if beta cell’s function was also enhanced in PHC-C6 and C-10 treatments.

5, In the Result section, the authors use blood glucose level as the indicator of the STZ and HFD model which is insufficient. It is suggested to also include other surrogate markers, for example, IPGTT/OGTT, and insulin levels to illustrate this model was successful.

6, Please include individual points of measuring parameters together with mean and SD in the bar chart.

7, In the Result section, line 253, the authors described ‘Metformin (70 mg/ Kg) showed significant reduction (***p<0.001) in cardiac collagen content.’ However, according to Fig 6, metformin didn’t decrease much.

 

Minor comments:

1, English writing needs polishing. Some descriptions lack clarity. For example, in the Result section, line ‘174’, the statement ‘The results of D-HFD-C group were…’ did not refer to the exact index.  

2, In fig. 2, the olive oil and PHC-C6, 10 groups still had a difference from the control group. Please also include the comparison between PHC-C6/10 with NC group.

3, In line’198’, the author used ‘test kits’ to analyze ALT, CK-MB and LDH. Please illustrate the detailed methods in the Method section.

4, The contents of lines ’241-248’ and lines ’256-274’ should be moved into the Method section.

Author Response

Please find attached below the response to reviewer's comments.

Author Response File: Author Response.docx

Reviewer 3 Report

1. References are missing in the introduction. The first reference is in row 61. Is all the information from rows 40-61 included in that article? I suggest that more Ref is included in the entire manuscript.

2. In rows 64-70, you refer to your own study in 2020 but the name of the first author is spelled incorrectly. And it´s not possible to open the link with the article.

3. The adult body weight for male Sprague-Dawley rats is 450-520 grams.          4-weeks old rats can´t be considered adults. Not until 8-10 weeks.

4. Why did you only use male rats, and not both male and female?

5. What did you inject the normal control group with when the other 6 groups were injected with STZ? On day 13, were they also abstained from eating for the time being and given a solitary intraperitoneal infusion of any control substance?

6. You write that "The carvedilol being cardioprotective standard, its alone treatment group reflected significant results (*p<0.05) when compared with D-HFD-C group as shown in fig 1.1 and fig 1.2." However, I can´t see any difference. Is it possible that you can point it out more clear in the figure?

7. Food Efficiency Ratio has to be explained.

8. Heart weight / body weight ratio could be written on the Y-axis.

9. I suggest that the Material and methods are written in the appropriate section and not in the result section.

10. Since you could conclude by this research that the polyherbal combination comprising T. cordifolia, W.somnifera, and B. diffusa plant extracts based in Olive Oil was found to have antidiabetic, antioxidant, and plays a prominent role in attenuation of diabetes-induced cardiomyopathy and associated anomalies; why don´t write this in your title?

 

Author Response

Please find attached below the response to reviewer's comments.

Author Response File: Author Response.docx

Round 2

Reviewer 1 Report

The authors have addressed majority of my concerns.

Reviewer 2 Report

My issues are well solved. The current version is decent for publication.