Journal of Functional Foods

Peer Review Report

Manuscript title: Arthrospira (Spirulina) platensis supplementation prevents
contractile reactivity damage in obese rats fed a hypercaloric diet by positive
modulating the Rho-A/Rho-kinase pathway, inflammation and oxidative stress.
Ms. Ref. No.: JFF-D-23-01487

Reviewer comments

In fact, the research idea is valuable. Moreover, the research assumptions were
evaluated in a scientific and adequate manner; however, there are some mistakes
and inquiries should be responded.

- Mistake correction:
Part/ Section Page/Line Correct from. To.
Abstract 33, 39 PA AP
Abstract 33 algae alga
may be associated with
may be associated with the
the presence of acid
Abstract 43 presence of gamma linolenic
gamma linolenic acid
acid (GLA)
(GLA)
Introduction 56, 57 ISMAIL et al., 2017 Ismail et al., 2015
Roval Manipulation Roval Manipulation
Materials and Page 9/34 Lines 36,
Pharmacy (João Pessoa), Pharmacy (João Pessoa,
methods 37
Paraiba, Brazil) Paraiba, Brazil)
Materials and Page 9/34 Lines 40, Animals and experiments Animals and experimental
methods 41 groups groups
purchased from the were purchased from the
Materials and Page 9/34 Lines 45,
Animal Production Unit Animal Production Unit
methods 46
(APU) (APU)
Materials and Page 9/34 Lines 50, with ad libitum access to with ad libitum access to
methods 51 water, water,
Materials and Page 10/34 Lines 14,
VET TEV
methods 15
Materials and modern chemistry
Page 10/34 Line 29 Modern Chemistry (Brazil)
methods (Brazil)
Materials and
Page 10/34 Line 33 nuclear (Brazil) Nuclear (Brazil)
methods
Materials and
Page 10/34 Line 51 was purchased were purchased
methods

1
Materials and
Page 11/34 Line 42 ad libitum ad libitum
methods

Materials and The ileum was assembled The ileum was assembled as
Page 12/34 Line 9
methods as described in item 3.1.1. described in item 3.2.1.

In the 1H NMR spectrum In the 1H NMR spectrum

Page 13/34 Lines 45,
Results (Fig. 1) was possible (Fig. 1), it was possible to
46
observed signals observe signals

Page 15/34 Lines 25, when compared to its

Results when compared to yours
26 absence

when compared to its when compared to its

Page 15/34 Lines 33,
Results absence. inhibitor absence (Emax = 63.9 ±
34, 35
(Emax = 63.9 ± 0.9%). 0.9%).

At the captions of all HCD = fed a hypercaloric HCD = hypercaloric diet

Results
figures and tables diet group group

Results Inside all tables pCE50 pEC50

in rats that consumed the
Results Page 17/34 Lines 1, 2 hypercaloric diet of the in rats from the HCD group
HCD group

Page 18/34 Lines 22, in rats fed the standard in rats from the SD
Results
23, 24 diet of the SD group, group,

in rats that consumed the

Page 18/34 Lines 31.
Results hypercaloric diet of the in rats from the HCD group
32, 33
HCD group

Page 18/34 Lines 36,

Results (Emax = 32 7 ± 7.5%), (Emax = 32.7 ± 7.5%),
37

(pEC50 = 6.2 ± 0.1) versus

Page 18/34 Lines 38, (pEC50 = 6.2 ± 0.1) (pEC50 = 6.6 ± 0.1) in its
Results
39 (Table 3, Fig. 6B, n = 5). absence (Table 3, Fig. 6B, n
= 5).

2
 In rats fed a hypercaloric
diet and supplemented
Page 18/34 Lines 40, In rats from the HCD+AP25
Results with 25 mg/kg of A.
41, 42 group
platensis in the
HCD+AP25 group
Interestingly, the Interestingly, the contractile
contractile potency of potency of CCh was reduced
CCh was reduced in the in the presence of the COX
Page 18/34 Lines 45, presence of inhibitor versus in its
Results
46, 47, 48 the COX inhibitor absence (pEC50 = 5.2 ± 0.04
(pEC50 = 7.2 ± 0.1; and pEC50 = 6.2 ± 0.1,
pEC50 = 5.2 ± 0.04) respectively) (Table 3, Fig.
(Table 3, Fig. 6B, n = 5). 6B, n = 5).
In rats fed a hypercaloric In rats from the HCD+AP25
diet and supplemented group
Page 19/34 Lines 11,
Results with 25 mg/kg of A.
12, 13
platensis in the
HCD+AP25 group
Similarly, the contractile Similarly, the contractile
potency of CCh was also potency of CCh was also
reduced in the presence reduced in the presence of
Page 19/34 Lines 16,
Results of the NOX inhibitor the NOX inhibitor versus in
17, 18
(pEC50 = 6.2 ± 0.1; its absence (pEC50 = 4.8 ±
pEC50 = 4.8 ± 0.1). 0.1 and pEC50 = 6.2 ± 0.1,
respectively).
Results Page 20/34 Line 56 SOD minetic SOD mimetic
Results Page 21/34 Lines 5, 6 in the HCD group rats, in the HCD group,
when compared to the when compared to the
Page 21/34 Lines 16,
Results absence of this inhibitor absence of this inhibitor,
17
no change in potency with no change in potency
Arthrospira (Spirulina) Arthrospira (Spirulina)
Discussion Page 22/34 Line 13
platensis platensis
Page 22/34 Lines 18, A. platensis A. platensis
Discussion
19, 20
is a blue-green is a blue-green microalga
Page 22/34 Lines 25,
Discussion microalgae traditionally traditionally used
26
used
Page 22/34 Lines 36, (XIAOPENG, et al., (Xiaopeng et al., 2023)
Discussion
37 2023)
were possible attribution were possible attribution to
Page 22/34 Lines 49,
Discussion to GLA (Figure 3) with GLA (Figure 3) as the main
50, 51
main compound compound
Page 22/34 Lines 51, (JUBIE, et al. 2015) (Jubie et al., 2015)
Discussion
52
Discussion Page 23/34 Lines 1, 2 A. platensis A. platensis
Discussion Page 24/34 Line 13 PA AP

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 Discussion Page 24/34 Line 31 COON et al., 2015 COON et al., 2005
Page 24/34 Lines 58, SILVA; CERCHIARO; Silva et al., 2011).
Discussion
59 HONORIO, 2011;).
In SD group rats, In SD group, or In rats of
Discussion Page 26/34 Line 33
SD group,
Additionally, the HCD Additionally, the HCD
Discussion Page 26/34 Line 44
group rats, group,
Page 27/34 Lines 24, tempol was used, a SOD Tempol, a SOD mimetic,
Discussion
25, 26 mimetic was used
In the AP-supplemented In the HCD+AP25 group,
Discussion Page 27/34 Line 46 group of the HCD+AP25
group,
Discussion Page 27/34 Line 57 (RSH) (R-SH)

- In addition to:
1. Highlights: The points are short and expressive; however,
1.1. It is preferred to paraphrase the sentence No. 2 to be "Peptides derived from Arthrospira
have biological activity either in vitro or in vivo" or "Peptides derived from Arthrospira have
biological activity in vitro and in vivo".
1.2. I think "highlights" are short sentences express the most important findings of the work
and/or the mechanisms through which these findings occurred; however I see the sentence No.4
is nothing more than the idea of the study.

2. Abstract: Although abstract should not exceed 150 words; however, such abbreviations that
are unavoidable must be defined at their first mention, for example, CCh and NADPH. This is
because abstract must be able to stand alone.

3. Keywords: Keywords are recommended to be separated only by ";", and there is no need for
numbering. Also, authors are recommended to add one or two expressive keywords used for
widely indexing purposes, for example, Contractile reactivity, Rats, etc.

4. Introduction: The introduction already stated the objectives of the work and provide an
adequate background; however.
4.1. Most recent references are required.
4.2. In the first paragraph, Lines 7, 8, I think malnutrition may be the best alternative to
"inadequate intake of macro and micronutrients", as the concept "malnutrition" is more

4
comprehensive and the health risks resulted from obesity is related to both over- and under-
nutrition.

5. Materials and methods: Some inquiries are found as follows:

5.1. The authors explained the percentages of nutrients in the standard diet used. Which chemical
materials were used to provide these nutrients? What is the reference? This should be explained.
5.2. ≥20 % protein, is used in standard diet preparation for young animals at growth stage, while
the used animals (170-190 g) are adults. Explain why?
5.3. In methods section, subsection 3.2.1. Obtaining and preparing rat ileum segments, the entire
paragraph is attributed to the same reference (RADENKOVIC et al., 2006), so it is
recommended to mention it at the end, i.e. there is no need for repetition.
5.4. A lot of abbreviations were found all over the "Materials and methods" section without
previous inserting for their full terms. Revision is needed.

6. Results: Results are clear and concise; However,

6.1: In page 14/34, the authors inserted a figure for the structure of gamma linolenic acid (GLA).
If GLA was the predominant compound in the sample of the AP chloroform extract, this should
be explained at the end of the comment on the subsection 4.1. NMR analysis of chloroform
extract, page 13/34, with an indication to the number of the figure showing the structure of this
compound.

7. Discussion: The discussion already explored the significance of the work results; however,
7.1. Care must be taken to unify the abbreviations used to refer to the same term. For example
two abbreviations were used to refer to the same term "voltage-dependent calcium channels; CaV
and Ca V", page 23/34 Lines 22, 35.
7.2. A lot of abbreviations were found all over the "Discussion" section without previous
inserting for their full terms. Revision is needed.
7.3. Some abbreviations do not fit the full terms inserted just before. For example, NADPH is
the abbreviation for reduced nicotinamide adenine dinucleotide phosphate (not for nicotinamide
adenine dinucleotide phosphate), etc. Revision is needed.
7.4. There is no need to refer to figures in the discussion.

5
8. Conclusions:
8.1. There is no need to refer to figures in the conclusions
8.2. Fig. 9 is the same as the graphical abstract. So, there is no need for repetition.

9. References: Revision is recommended since the following:

9.1. The standard reference style of the journal should be followed in all references cited both in
the text and in the reference list. Moreover, a unified method must be followed.
9.2. According to the reference list, "FELIPPI, 2014" mentioned in the "introduction" section
lines 27, 28 should be corrected, as FELIPPI is not the only author.
9.3. About 14 references were missed in the reference list. They include the references:
(Lancet, 2016) mentioned in the "Introduction" section, lines 10, 11;
(BATISTA et al., 2018) mentioned in the "Introduction" section lines 38, 39;
(MILLS et al., 2001) mentioned in both the "Materials and methods" section page 12/34 lines
14, 15, and in the "Discussion" section page 23/34 lines 54, 55;
(CÔCO et al., 2016) mentioned in the "Materials and methods" section page 12/34 lines 36, 37;
(PEIXOTO et al., 2009) mentioned in both the "Materials and methods" section page 12/34
lines 37, 38, and in the "Discussion" section page 27/34 lines 25, 26;
(VIGNOZZI et al., 2006) mentioned in both the "Materials and methods" section page 12/34
lines 58, 59, and in the "Discussion" section page 24/34 line 44;
(XIAOPENG, et al., 2023) mentioned in the "Discussion" section page 22/34 lines 36, 37;
(YAN et al., 2012) mentioned in the "Discussion" section page 24/34 line 40;
(JOHNSON et al.., 2011) mentioned in the "Discussion" section page 25/34 lines 16, 17;
(XIANG et al., 2008) mentioned in the "Discussion" section page 25/34 line 31;
(HATANAKA et al., 2006) mentioned in the "Discussion" section page 27/34 line 11;
(FINKEL, 2001) mentioned in the "Discussion" section page 27/34 lines 58, 59;
(NAKASHIMA et al., 2002) mentioned in the "Discussion" section page 27/34 lines 60, 61;
(WANG et al., 2018) mentioned in the "Discussion" section page 28/34 line 1.
9.4. The information of some references was not complete.
9.5. The scientific method of writing references (e.g. starting with family name, etc.) was not
followed in the reference (SATISH RATTAN et al., 2010) mentioned in the text in the
"Discussion" section page 23/34 lines 49,50, 51, while it was inserted correctly in the reference
list.