Echinacea purpurea Alleviates Cyclophosphamide-Induced Immunosuppression in Mice

Round 1

Reviewer 1 Report

It is very well executed experiment. Utilizing natural products for immunomudulation is novel. The active extract from Echinacea purpurea was used. But, none of the possible alkaloids or terpenoids were mentioned as the chemical composition of this extract. A brief discussion about the extract chemical constituents will be very helpful to the reader. The experiments are very well executed especially the mouse model for this study.

Author Response

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Thanks for good review indication. 

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Reviewer 2 Report

1. Cyclophosphamide is a drug used mainly in cancer treatment. It can be used also after organ transplantations, having immunosuppressive activity. However, I think choice of cyclophosphamide for immunosuppressive studies is not good. Today, main immunosuppressive drugs, used worldwide are glucocorticosteroids. Therefore, Authors should study one or some of glucocorticosteroids, instead of anticancer cyclophosphamide. It will be more useful for science and patients.
2. In the article is lack of phytochemical analysis of Echinacea purpurea. The phytochemical composition differs significantly from the collection site, soil, sunlight, etc. Research without a thorough phytochemical analysis is pointless.
3. Methodology is poorly described; very often is lack of names of reagents, kits, producers, conditions of studies.

Author Response

I attached revision comments via Word file. 

Thanks for good review indication. 

Author Response File: Author Response.pdf

Reviewer 3 Report

The authors evaluated an extract of Echinacea purpurea and suggest that it could be used to improve health and immunity under conditions of immunosuppression.

The manuscript is well organized although

I suggest adding these manuscripts (see below) in the introduction to emphasize the importance of medicinal plants and their potential help for people's health.

Fernández J, Silván B, Entrialgo-Cadierno R,Villar  C.J, Capasso R., Uranga J. A., Lombó F, l Abalo R., Antiproliferative and palliative activity of flavonoids in colorectal cancer. Biomed Pharmacother. 2021; 143: 112241, doi.org/10.1016/j.biopha.2021.112241

Küpeli Akkol E, Genç Y, Karpuz B, Sobarzo-Sánchez E, Capasso R. Coumarins and

Coumarin-Related Compounds in Pharmacotherapy of Cancer. Cancers (Basel). 2020

Jul 19;12(7):1959.

The plant extracts should contain a type of standardization (i.e. fingerprint)

Do the authors have information on whether the individual constituents can have a synergistic effect?

In the Discussion, the Authors should highlight the possible clinical significance

Author Response

I attached revision comments via Word file. 

Thanks for good review indication. 

Author Response File: Author Response.pdf

Reviewer 4 Report

The authors investigated the effects of Echinaceae purpurea against immune suppression by the chemotherapeutic drug  cyclophosphamide in mice.

Is there in the literature studies that have investigated the immuno-enhance activity of other plants from Asteraceae family? If so, pleae include it in the introduction section.

The description of the preparation of EP extract is too superficial. Please include details that could turn it reproducible.

How have you chosen the doses 12,5, 25 and 50 mg/kg?

Which method of euthanasia have you used?

Why have you administered CP on days 10 as 12? Is there any reference for it? 

Why have you used Duncan´s post-hoc test? 

Author Response

I attached revision comments via Word file. 

Thanks for good review indication. 

Author Response File: Author Response.pdf

Round 2

Reviewer 2 Report

The authors explained why studied cyclophosphamide, with which I can partially agree. 

Unfortunately:

- is a lack of phytochemical analysis of Echinacea purpurea. The authors confirmed chicoric acid in approximately 2%, but it is not proper phytochemical analysis. In plant is only 1 compound??? What with the rest 98% of compounds? Such an analysis is worth nothing.

- Methodology is partially corrected, but generally all time is poorly described.