Background： Gender equality and equitable health have emerged as international trends in
recent years. Despite existing solutions for women's health issues, many needs remain
unmet. According to projections, the global menopausal population is expected to reach
1.2 billion by 2030, surpassing the prevalence of the most common chronic diseases.
Presently, solutions exist for common menopausal symptoms such as hot flashes, night
sweats, fatigue, irritability, and urinary incontinence. However, unresolved challenges
encompass psychological changes and monitoring systems. Therefore, this project aims to
develop biomarkers for menopause in women as a future monitoring system.
Most women enter menopause between the ages of 48 and 52. During this period, ovarian
follicles cease activity, leading to the one-year period following the appearance of
significant menstrual irregularities, known as the menopausal transition or perimenopause
(1). As ovarian function gradually declines and estrogen secretion diminishes, various
systemic changes occur, resulting in several menopause-related conditions such as
osteoporosis, cardiovascular diseases, and urinary system issues. The decline in estrogen
triggers a negative feedback loop, causing an increase in follicle-stimulating hormone
(FSH) secretion and a reduction in E2 (17β-estradiol) production (2). Currently,
menopausal hormone therapy (MHT) using estrogen can alleviate menopausal symptoms and
restore overall E2 levels but doesn't lower FSH to premenopausal levels (3-4). However, a
precise biomarker for monitoring menopause has not yet been established. Under estrogen
deficiency, mitochondrial morphology and function may be compromised, while estrogen
presence relates to reduced oxidative reactions, enhanced respiratory function, and
stable membrane properties (5). This project aims to provide a comprehensive view through
metabolomics to identify biomarkers within mitochondria that can be used for menopause
monitoring.
Study Design： This study is a cross-sectional study. A total of 100 women will be
included and grouped into three categories: 25 cases of reproductive period, 50 cases of
perimenopausal transition period, and 25 cases of postmenopausal period. The peripheral
blood will be collected. Following mitochondrial extraction, both qualitative and
quantitative analyses are conducted. High-confidence protein markers are identified as
candidate molecules using combined data from RNA-seq and LC/MS/MS. These candidate
markers will undergo further validation testing.
Methods： After Ficoll separation, peripheral blood will be divided into mononuclear
cells, plasma, and red blood cells. Further extraction of mitochondria will be carried
out from both mononuclear cells and plasma. Quantitative experiments involve hippocampal
measurements of metabolic energy and JC-1 assessment of mitochondrial health, while
mitochondrial DNA copy number will be evaluated using digital PCR. Qualitative
experimental analysis employs Western blotting to assess markers such as cytochrome c and
HSP60 for mitochondria and PCNA for nuclear proteins. Candidate biomarker validation
experiments use combined data from RNA-seq and LC/MS/MS to identify high-confidence
protein markers as candidates, which are further validated for feasibility using the
ELISA method.
Effect：
Understand the changes in mitochondrial quality and quantity during the
perimenopausal transition in women.
Develop monitoring indicators for the perimenopausal transition in women.