Effects of Water Extract from Artemisia argyi Leaves on LPS-Induced Mastitis in Mice

Round 1

Reviewer 1 Report

With the help of evidence-based experimental biology, the prospects for the development of medicinal preparations from plants that have long been used in traditional Chinese medicine are substantiated in the work, proposed for publication . The problem raised is undoubtedly of interest. However, there are several questions and comments on the work done.

1. At the very beginning of the article, the abbreviation ALE is used (line 23), the decoding in the text is given much further (line 81). I believe that a decoding should be given already at the first use of this abbreviation.
2. Line 137-138: “Total RNA extraction kit was used to isolate proteins from homogenates…”. Perhaps they meant "to isolate RNA"?
3. Line 158: "LPS infection of the breast usually causes clinical mastitis." I consider this proposal incorrect, since the term "infection" is applicable to pathogens - bacteria, viruses, etc.
4. Lines 223-225: “As shown in Figure 5, the expression levels of IL6, TNFα and IL1β in breast tissues were significantly increased by the addition of LPS (p < 0.01). Addition of ALE also significantly increased their expression levels (p < 0.01)…” indeed ALE increase the expression of these cytokines, but dozens of times less prominent compared to LPS, so it is incorrect to write “significantly increased” in both cases.
5. And further lines 225-226 “…but ALE significantly down-regulated the expression level of LPS (p < 0.01)” Perhaps they meant “expression level of IL6, TNFα and IL1β on the background of LPS stimulation”?
6. Why were non-lactating animals used in the development of a mouse model of mastitis?
7. Why were such high concentrations of ALE (10 mg/mL) used in the work? In their other work, to which the authors refer, it was shown that ALE at a concentration of 1 μg/ml has the strongest cytotoxic effect, cell survival is less than 20%.
8. LPS concentration is also unreasonably high (0.2 mg/mL). Mouse models of mastitis have previously been described in which LPS was administered at a concentration of 20 ng/μL (Xiao HB, Wang CR, Liu ZK, Wang JY. LPS induces pro-inflammatory response in mastitis mice and mammary epithelial cells: Possible involvement of NF-κB signaling and OPN. Pathol Biol (Paris). 2015 Feb;63(1):11-6. doi: 10.1016/j.patbio.2014.10.005. Epub 2014 Nov 4. PMID: 25468491.)
9. Why do the authors believe that substances that suppress the expression of inflammatory mediators are a good alternative to antibiotics for mastitis? Inflammation is the immune system's way of fighting infectious agents. If plant extracts, along with an anti-inflammatory effect, also have a bacteriostatic or bactericidal effect, then this must be demonstrated in experiments.

Author Response

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Reviewer 2 Report

I appreciate the idea of the manuscript. However, I recommend introducing some changes as follows:

 

Line 21: Remove one "." after hotspot

Please define the abbreviation ALE in the abstract

Line 42-43- I don`t understand the division based of "former" and "latter". Please clarify

Line 91 Preparation ?

Table 1 please provide explanation for CK, ALE etc. The table shoud stand alone.

Line 117- maybe change "kill" for "euthanised"

Lines 153- 155 - delete

Line 248 - space befor citation

Lines 328 -331 - have you used bovines in the study ?

lines 333-334 - correct "material"

Author Response

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Reviewer 3 Report

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Comments for author File: Comments.pdf

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Round 2

Reviewer 3 Report

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Comments for author File: Comments.pdf

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