Gastroprotective, Biochemical and Acute Toxicity Effects of Papaver decaisnei against Ethanol-Induced Gastric Ulcers in Rats

Round 1

Reviewer 1 Report

The experiments were carried out properly. However, some questions should be clarified before the paper published.

1.     How were the samples (200mg/kg and 400mg/kg) prepared for intragastric administration of rats.

2.     Negative controls are missing in immunohistochemistry.

3.     In Line 366-367,“Previous toxicological studies…”the author should cite at least 2 literatures to illustrate the problem.

Author Response

Response to Reviewers Comments

 

The manuscript has been thoroughly revised according to the referees’ comments (point-by-point) and changes are highlighted in different colors. Our responses to the referees’ comments are detailed as follows.

Reviewer 1 (changes are in light yellow):

The experiments were carried out properly. However, some questions should be clarified before the paper published.

1. How were the samples (200 mg/kg and 400 mg/kg) prepared for intragastric administration of rats?

Answer: According to weight of rate, for example for low dose (200mg/Kg) we prepared 200mg of dry extract dissolved in 5 ml of CMC (carboxymethylcellulose). feeding 1ml of sample for each Rate. Also for higher dose 400mg of dry extract was dissolved in 5 ml of CMC. Of Note: 5 Rate totally equal 1kg each Rate 200g. and for prepare CMC as following: 0.5% carboxymethylcellulose was prepared by adding 5 grams of absolute carboxylmethyl cellulose into 1L in Normal Saline (0.9% sodium chloride). At first, we put (250 ml) normal saline in volumetric flask (1L) then we use beaker (1L) to dissolved 5g of powder CMC In (500ml) of N.S solution, to make the dissolving the CMC easier. After that both of solutions were mixed in volumetric flask (1L) as a result the volume became 750ml. Finally, N.S was added to the volumetric till the solution volume reach 1000ml. for acute toxicity and gastric ulcer experiments the same procedure was used to prepare the CMC solution (negative control).

2. Negative controls are missing in immunohistochemistry.

Answer: Negative control added to this section.

3. In Line 366-367, “Previous toxicological studies” the author should cite at least 2 literatures to illustrate the problem.

   Answer: More articles cited on that regards.

 

Author Response File: Author Response.docx

Reviewer 2 Report

The work presented here is well designed and presents the information correctly.

 

Doubts. Since they demonstrate the protective effect of the Papaver plant, at the histological and biochemical level.

I would like to suggest that the discussion and conclusions be extended in relation to the possible effect that Papaver plant would have on the regulation of oxidative stress and inflammation by regulating the transcriptional factor NRF2.

Since it has been shown that this factor regulates both the inflammatory process and oxidative stress.

 

Thank you

Author Response

Reviewer 2 (changes are in light green):

Doubts. Since they demonstrate the protective effect of the Papaver plant, at the histological and biochemical level.

I would like to suggest that the discussion and conclusions be extended in relation to the possible effect that Papaver plant would have on the regulation of oxidative stress and inflammation by regulating the transcriptional factor NRF2.

Since it has been shown that this factor regulates both the inflammatory process and oxidative stress.

Oxidative stress has been known as a series cause of inflammation, which defined as an imbalance in ROS (superoxide, hydroxyl free radicals, and hydrogen peroxide), which also lowers antioxidant enzyme productions that fights free radicals [32]. Antiradical enzymes such as phase 2 antioxidant induced enzymes and cell protective genes (glutamate-cysteine ligase catalytic (GCLC), heme oxygenase-1 (HO-1), and NAD(P)H quinone oxidoreductase 1 (NQO1)) were activated via nuclear factor erythroid-2-related factor 2 (Nrf2), which has an important role in cell protection from free radicals and inflammations [36-38]. Nrf2 is dimerized with Maf small-sized proteins in the nucleus and joins cis-regulatory with the antioxidant response-element to stimulate the transcriptional expression. This action could be down-regulated by numerous genes, such as NQO1 and HO-1 as scientists have been clarified [38]. Furthermore, the NF-κB signaling considered as a suppressor of the antioxidant actions due to its inhibitory effects on the Nrf2-Keap1 mechanism via the p65 and Keap1 interactions [39]. Thus, establishing new anti-inflammatory Nrf2 signaling is crucial to enhance cell’s antioxidant defense systems, due to its potentials in up-regulation of cell protection genes (HO-1, NQO1, and GCLC) [38]. The current study showed significant antioxidant potentials of P. decaisnei, which could be through activation of antioxidant genes mediated by Nrf2. Similarly, a recent study has reported the isoquinoline alkaloids profile and the antioxidant potentials of P. nudicaule via mechanism of the Nrf2-mediated stimulation of antiradical genes. While, a notable antioxidant actions of P. rhoeas extracts were explained via up-regulation of NQO1 without Nrf2 mediation, possibly via suppressing NF-κB [40].

Author Response File: Author Response.docx

Reviewer 3 Report

There are a few notes on writing the article.
1) The reviewer is not sure that fig. 1 is essential for understanding the mechanism of anti-ulcerogenic actions of Papaver decaisnei. However, in any case, a reference to this figure should be made in the text.

2) The numbering of subsections in section 2 is broken, starting with subsection 2.4.

3) It is not clear which regions are indicated by the arrows in Fig. 3D.

Author Response

 

Response to Reviewers Comments

 

Reviewer 3 (changes are in turquoise):

There are a few notes on writing the article.
1) The reviewer is not sure that fig. 1 is essential for understanding the mechanism of anti-ulcerogenic actions of Papaver decaisnei. However, in any case, a reference to this figure should be made in the text.

Answer: Thanks for your comment, yes its essential, I mentioned (Fig 1) in the text from section 2.1

 

2) The numbering of subsections in section 2 is broken, starting with subsection 2.4.

 

Answer: The section 2, totally have been corrected.

 

3) It is not clear which regions are indicated by the arrows in Fig. 3D.

 

Answer: The region has been corrected.

Author Response File: Author Response.docx

Round 2

Reviewer 1 Report

The authors have revised the manuscript accordingly.