Title: Neuregulin1 ameliorates metabolic dysfunction-associated fatty liver disease via the
ERK/SIRT1 signaling pathways
Version: 2 Date: 15 Sep 2024
Reviewer's report:
Reviewers' comments
Title:Neuregulin1 ameliorates nonalcoholic fatty liver disease via the ERK/SIRT1 signaling
pathways
Comments:
Neuregulin (NRG) family is involved in energy metabolism, among which NRG1 is a
neuregulin has been proved to upregulate the phosphorylation of ErbB3 in NAFLD cells.
This study aimed to investigate the role of NRG1 in the pathogenesis of hepatic steatosis
and steatohepatitis.HepG2 cells were exposed to oleic acid and palmitic acid to establish a
cellular model of NAFLD. NAFLD animal model was established by feeding C57BL/6J mice
with high-fat diet. H&E staining, oil red staining, and biochemical tests were used to evaluate
the models. The results  showed that #The expressions of NRG1 in NAFLD cell and animal
models were significantly lower than the control group. The intervention of NRG1 increased
its expressions. After the intervention of ERK inhibitor PD98059, no significant change was
observed in vivo, while its expression decreased significantly in vitro. #NRG1 ameliorates
hepatic steatosis, enhances cell viability, reduces cell apoptosis, and attenuates liver injury both
in vitro and in vivo. #After NRG1 intervention, the expressions of ERBB2, ERRBB3, p-ERK1/2,
SIRT1 and p-FOXO1 in NAFLD cells and liver tissues of NAFLD mice were significantly
increased. Furthermore, the LC3II/I ratio was increased, while the expression of SREBP1c was
decreased. The data above suggest  that  NRG1 might play a protective role in the pathogenesis
of NAFLD by activating the downstream ERK1/2 through the ErbB2-ErbB3, which promotes
the expression of SIRT1 and autophagy.
It is a topic of interest to the researchers in the related areas #but the paper needs large
improvements before acceptance for publication.My detailed comments are as follows:
1.the introduction ,materials and methods  in the paper work  well ,Cell culture
and treatments,nimal models,Oil Red O staining, Cell viability assay,Apoptosis analysis,
Intracellular ROS,Measurement of serum inflammatory factors,and  Western blotting.
2. Results are good and the resolutions of the figures are high, but the results got in the
experiments should be described objectively,not conclusions were made;In addition,the part of
dicussion is not well discussed combined with results and references and should make some
modifications.
3.The language is not fluent ,suggesting that it should be edited by an english native editor.
4.There are 19.4%  references which are not up-to-date,references of the last 10 years should be
cited,please cite last 10 years references ,especially references for the last 5 years.
Pelase make large revisions, espically in the part of results,discussion references  and languge-
editing. After making large  revisions,the paper may be considered for publication.
Title: Neuregulin1 ameliorates metabolic dysfunction-associated fatty liver disease via the
ERK/SIRT1 signaling pathways
Version: 2 Date: 15 Sep 2024
Reviewer's report:
Reviewers' comments
Title:Neuregulin1 ameliorates nonalcoholic fatty liver disease via the ERK/SIRT1 signaling
pathways
Comments:
Neuregulin (NRG) family is involved in energy metabolism, among which NRG1 is a
neuregulin has been proved to upregulate the phosphorylation of ErbB3 in NAFLD cells.
This study aimed to investigate the role of NRG1 in the pathogenesis of hepatic steatosis
and steatohepatitis.HepG2 cells were exposed to oleic acid and palmitic acid to establish a
cellular model of NAFLD. NAFLD animal model was established by feeding C57BL/6J mice
with high-fat diet. H&E staining, oil red staining, and biochemical tests were used to evaluate
the models. The results  showed that #The expressions of NRG1 in NAFLD cell and animal
models were significantly lower than the control group. The intervention of NRG1 increased
its expressions. After the intervention of ERK inhibitor PD98059, no significant change was
observed in vivo, while its expression decreased significantly in vitro. #NRG1 ameliorates
hepatic steatosis, enhances cell viability, reduces cell apoptosis, and attenuates liver injury both
in vitro and in vivo. #After NRG1 intervention, the expressions of ERBB2, ERRBB3, p-ERK1/2,
SIRT1 and p-FOXO1 in NAFLD cells and liver tissues of NAFLD mice were significantly
increased. Furthermore, the LC3II/I ratio was increased, while the expression of SREBP1c was
decreased. The data above suggest  that  NRG1 might play a protective role in the pathogenesis
of NAFLD by activating the downstream ERK1/2 through the ErbB2-ErbB3, which promotes
the expression of SIRT1 and autophagy.
It is a topic of interest to the researchers in the related areas #but the paper needs large
improvements before acceptance for publication.My detailed comments are as follows:
1.the introduction ,materials and methods  in the paper work  well ,Cell culture
and treatments,nimal models,Oil Red O staining, Cell viability assay,Apoptosis analysis,
Intracellular ROS,Measurement of serum inflammatory factors,and  Western blotting.
2. Results are good and the resolutions of the figures are high, but the results got in the
experiments should be described objectively,not conclusions were made;In addition,the part of
dicussion is not well discussed combined with results and references and should make some
modifications.
3.The language is not fluent ,suggesting that it should be edited by an english native editor.
4.There are 19.4%  references which are not up-to-date,references of the last 10 years should be
cited,please cite last 10 years references ,especially references for the last 5 years.
Pelase make large revisions, espically in the part of results,discussion references  and languge-
editing. After making large  revisions,the paper may be considered for publication.
Please review this paper.
user
Title: Neuregulin1 ameliorates metabolic dysfunction-associated fatty liver disease via the
ERK/SIRT1 signaling pathways
Version: 2 Date: 15 Sep 2024
Reviewer's report:
Reviewers' comments
Title:Neuregulin1 ameliorates nonalcoholic fatty liver disease via the ERK/SIRT1 signaling
pathways
Comments:
Neuregulin (NRG) family is involved in energy metabolism, among which NRG1 is a
neuregulin has been proved to upregulate the phosphorylation of ErbB3 in NAFLD cells.
This study aimed to investigate the role of NRG1 in the pathogenesis of hepatic steatosis
and steatohepatitis.HepG2 cells were exposed to oleic acid and palmitic acid to establish a
cellular model of NAFLD. NAFLD animal model was established by feeding C57BL/6J mice
with high-fat diet. H&E staining, oil red staining, and biochemical tests were used to evaluate
the models. The results  showed that #The expressions of NRG1 in NAFLD cell and animal
models were significantly lower than the control group. The intervention of NRG1 increased
its expressions. After the intervention of ERK inhibitor PD98059, no significant change was
observed in vivo, while its expression decreased significantly in vitro. #NRG1 ameliorates
hepatic steatosis, enhances cell viability, reduces cell apoptosis, and attenuates liver injury both
in vitro and in vivo. #After NRG1 intervention, the expressions of ERBB2, ERRBB3, p-ERK1/2,
SIRT1 and p-FOXO1 in NAFLD cells and liver tissues of NAFLD mice were significantly
increased. Furthermore, the LC3II/I ratio was increased, while the expression of SREBP1c was
decreased. The data above suggest  that  NRG1 might play a protective role in the pathogenesis
of NAFLD by activating the downstream ERK1/2 through the ErbB2-ErbB3, which promotes
the expression of SIRT1 and autophagy.
It is a topic of interest to the researchers in the related areas #but the paper needs large
improvements before acceptance for publication.My detailed comments are as follows:
1.the introduction ,materials and methods  in the paper work  well ,Cell culture
and treatments,nimal models,Oil Red O staining, Cell viability assay,Apoptosis analysis,
Intracellular ROS,Measurement of serum inflammatory factors,and  Western blotting.
2. Results are good and the resolutions of the figures are high, but the results got in the
experiments should be described objectively,not conclusions were made;In addition,the part of
dicussion is not well discussed combined with results and references and should make some
modifications.
3.The language is not fluent ,suggesting that it should be edited by an english native editor.
4.There are 19.4%  references which are not up-to-date,references of the last 10 years should be
cited,please cite last 10 years references ,especially references for the last 5 years.
Pelase make large revisions, espically in the part of results,discussion references  and languge-
editing. After making large  revisions,the paper may be considered for publication.
assistant