Title: Associations of LH and FSH with reproductive hormones depending on each stage of the
menopausal transition
Version: 1 Date: 23 Oct 2022
Reviewer's report:
The authors aimed to investigate the associations between gonadotropins LH and FSH and
several sex hormones in women of different reproductive/menopausal stages. The study was a
cross-sectional study of  173 relatively healthy Japanese women.
Abstract: please add information on cross-sectional design and the use of STRAW+10. In
conclusions, the last sentence "The associations between...." is not really related to anything in
the abstract.  Please add relevant leading information to this statement.
Introduction: Row 93-94: "...with changes in reproductive hormones." What changes are meant,
levels or actions?
Row 105-107:  The associations were investigated also before menopausal transition and after
FMP, not only during menopausal transition.
Overall, a simple figure illustrating the delta-4 and 5-steroidogenic pathways would make the
text more easy to follow.
Statistical analysis: Please provide explanation why Kruskal-Wallis test and Spearman test were
used. Was normality  tested?
Results: Table 1: please report all measured hormones (estradiol, testosterone etc) in all  six
groups. In addition, I would also like know in more detail about the health status of the
participants since some of them had dyslipidemia etc.
Row 153: please avoid using the term "changes" and "increase" due to the cross-sectional nature
of the study and be consistent on the use of the term "menopausal transition" or define in
more detail the terminology you will use.  Earlier you stated that only groups C and D were in
menopausal transition according to STRAW + 10.
Row 155/Figure 1: I would recommend changing ** to some other sign, as this is very intuative
to being more significant than *.
Table 2 and 3: why is cortisol here? It is not mentioned in methods nor discussed anywhere.
Rows 168-179: some of the reported correlations are weak (<0.4). Overall, a scatterplot of
separate associations  would be more informative for the reader to evaluate  whether the
association is real.
Discussion: rows 191-193: You state: "Through an increase in FSH level, a relationship in which
the balance between LH and FSH was maintained in Group A was attenuated, and the shift was
beginning toward the delta-5 pathway." The terminology used suggests that you have somehow
measured pathway activity, but you have not. Please use more accurate language.
Row 193: "In addition, stimulation of DHEAS production .." where is this stimulation occuring?
Rows 196-197: Basically repeats the next sentence.  Why is fibrosis important to mention?
Rows 200-202, reference 25: is this a valid reference for reporting  ovarian steroidogenesis if the
study was done with tumour cells? Also the PCOS-reference, is it valid to clarify your results
since the women have a medical condition (PCOS)?
Rows 210-212: here also you suggest that you have measured the activity of the route (enzymes) .
I would recommend being more careful with the conclusions.
Rows 219-220: "Due to the increase in LH...." please provide a reference for this statement.
Rows 223-225: "As well as DHEAS...." please provide a reference for this statement.
Rows 231-233: I don't understand what this sentence is trying to say.
Rows 234-235: "The reason is conversion...". This was not investigated in enzyme level in your
study. Please be more careful with you words or provide a reference.
Rows 235-237: the sentence is difficult to read, please rephrase.
Rows 242-243: You state "perimenopausal women" and "older women". Please provide the
participant age and menopausal status of both studies, so the reader knows what you really talk
about.
Overall, when talking about steroidogenesis, please add the details of where it is occuring  or
studied (ovaries, adipose tissue, adrenal gland, muscle....) for clarity.  Additionally, now you
have not discussed much on the effects of measured sex hormones for health, only LH and FSH.
I am not quite sure whether the authors think it is the gonadotropins that affect health or the sex
hormones? This should be made more clear to understand why is it important to investigate the
associations between gonadotropins and sex hormones.