Title: Dietary intervention reverses molecular markers of hepatocellular senescence in the GAN
diet-induced obese and biopsy-confirmed mouse model of NASH
Version: 1 Date: 06 Dec 2023
Reviewer's report:
The authors provided interesting findings on the beneficial effect of dietary intervention for
the amelioration of diet-induced NASH model. ALthough the results are interesting, several
experiments may be necessary to clearly strengthen their findings as shown below:
1. While the authors performed a few experiments with regard to hepatocellular senescence, p21
positive cell area and SASP would not be sufficient to fully address what the authors intended
to demonstrate in the title of rhis manuscript. At least b-galactosidase staining or other types of
senescence markers (e.g., Lamin B1, gammaH2AX, etc.) need to be further provided.
1-1. Although the authors performed RNA sequencing using livers obtained from experimental
mice, the authors may consider examining the protein expression of several key molecules so
as to fully validate their findings.
2. It is advised to present additional in vivo data in relation to glucose metabolism (e.g., plasma
glucose, plasma insulin, insulin sensitivity markers in representative organs), so the effect of
dietary intervention on NASH can be further demonstrated.
3. How the weight of adipose tissue was affected? Can this change be attributable to overall
reduction of body weight upon dietary intervention in those mice?
4. In order to prove subtle change of fibrosis markers upon dietary intervention, the authors may
perform additional experiments to analyze fibrotic changes, including hydroxyproline contents,
Masson's Trichrome staining, etc.
4-1. While the overall fibrosis score did not markedly change upon  8 or 16 weeks of dietary
intervention, alpha-SMA or Col1a1 positive area were significantly reduced. Any explain for
this discrepancy?
4-2. Can the authors examine whether the activity and/or the population of hepatic stellate cells
were markedly affected by dietary intervention?
Title: Dietary intervention reverses molecular markers of hepatocellular senescence in the GAN
diet-induced obese and biopsy-confirmed mouse model of NASH
Version: 1 Date: 06 Dec 2023
Reviewer's report:
The authors provided interesting findings on the beneficial effect of dietary intervention for
the amelioration of diet-induced NASH model. ALthough the results are interesting, several
experiments may be necessary to clearly strengthen their findings as shown below:
1. While the authors performed a few experiments with regard to hepatocellular senescence, p21
positive cell area and SASP would not be sufficient to fully address what the authors intended
to demonstrate in the title of rhis manuscript. At least b-galactosidase staining or other types of
senescence markers (e.g., Lamin B1, gammaH2AX, etc.) need to be further provided.
1-1. Although the authors performed RNA sequencing using livers obtained from experimental
mice, the authors may consider examining the protein expression of several key molecules so
as to fully validate their findings.
2. It is advised to present additional in vivo data in relation to glucose metabolism (e.g., plasma
glucose, plasma insulin, insulin sensitivity markers in representative organs), so the effect of
dietary intervention on NASH can be further demonstrated.
3. How the weight of adipose tissue was affected? Can this change be attributable to overall
reduction of body weight upon dietary intervention in those mice?
4. In order to prove subtle change of fibrosis markers upon dietary intervention, the authors may
perform additional experiments to analyze fibrotic changes, including hydroxyproline contents,
Masson's Trichrome staining, etc.
4-1. While the overall fibrosis score did not markedly change upon  8 or 16 weeks of dietary
intervention, alpha-SMA or Col1a1 positive area were significantly reduced. Any explain for
this discrepancy?
4-2. Can the authors examine whether the activity and/or the population of hepatic stellate cells
were markedly affected by dietary intervention?
Please review this paper.
user
Title: Dietary intervention reverses molecular markers of hepatocellular senescence in the GAN
diet-induced obese and biopsy-confirmed mouse model of NASH
Version: 1 Date: 06 Dec 2023
Reviewer's report:
The authors provided interesting findings on the beneficial effect of dietary intervention for
the amelioration of diet-induced NASH model. ALthough the results are interesting, several
experiments may be necessary to clearly strengthen their findings as shown below:
1. While the authors performed a few experiments with regard to hepatocellular senescence, p21
positive cell area and SASP would not be sufficient to fully address what the authors intended
to demonstrate in the title of rhis manuscript. At least b-galactosidase staining or other types of
senescence markers (e.g., Lamin B1, gammaH2AX, etc.) need to be further provided.
1-1. Although the authors performed RNA sequencing using livers obtained from experimental
mice, the authors may consider examining the protein expression of several key molecules so
as to fully validate their findings.
2. It is advised to present additional in vivo data in relation to glucose metabolism (e.g., plasma
glucose, plasma insulin, insulin sensitivity markers in representative organs), so the effect of
dietary intervention on NASH can be further demonstrated.
3. How the weight of adipose tissue was affected? Can this change be attributable to overall
reduction of body weight upon dietary intervention in those mice?
4. In order to prove subtle change of fibrosis markers upon dietary intervention, the authors may
perform additional experiments to analyze fibrotic changes, including hydroxyproline contents,
Masson's Trichrome staining, etc.
4-1. While the overall fibrosis score did not markedly change upon  8 or 16 weeks of dietary
intervention, alpha-SMA or Col1a1 positive area were significantly reduced. Any explain for
this discrepancy?
4-2. Can the authors examine whether the activity and/or the population of hepatic stellate cells
were markedly affected by dietary intervention?
assistant