Iron-deficiencyIronFEMMENCT04458662
Iron and Muscular Damage: FEmale Metabolism and Menstrual Cycle During Exercise
An observational study of Iron-deficiency and Inflammation, run by Universidad Politecnica de Madrid at 1 site in Spain.
Trial summary
This project is an observational controlled randomized counterbalance study. One hundred and three physically active and healthy women were selected to participate in the IronFEMME Study, of which 57 were eumenorrheic, 30 were oral contraceptive users (OCP) and 16 were postmenopausal women. Read more about this study
- Who is it for?
- Ages 18 to 60 years, women onlyHealthy volunteers can take part
- Conditions the study is for
- Iron-deficiency, Inflammation, Exercise Induced Iron-deficiency
- What is being tested?
- Interval running protocol / eccentric-based resistance exercise protocol
- Type of treatment
- Pill or oral medicine, Surgery or procedure
- How many people
- 103 participants
- Where
- 1 siteSpain
- Study dates
- 1 Jan 2017to 1 Jun 2020
- How long will I take part?
- 2 Months
- Phase
- Observational
Key requirements
Who can take part, as listed by the sponsor. Only the study team can confirm whether you are eligible, so take this list to your doctor.
- Ages 18 to 60 years
- Women only
- Healthy volunteers accepted
- Adult
Study population: Physically active and healthy women. Study I, 37 eumenorrheic women (30.0±6.3 yrs; 59.8±15.7 kg; 163.7±6.3 cm): endurance training (ET) experience of 7.7 yrs and training volume of 5.5±0.9 h/week. Study II, 20 eumenorrheic women (28.8±6.2 yrs; 57.5±13.8 kg; 163.9±6.4 cm): strength training (ST) experience of 6.4±4.1 yrs and training volume of 7.5±2.1 h/week. Eumenorrheic women selected to participate in Study I and II were different, whereas oral contraceptive users and postmenopausal women participated in both studies. 30 oral contraceptive users (25.1±4.3 yrs; 56.2±10.9 kg; 163.1±5.5 cm): ET experience of 7.3±5.5 yrs and training volume of 3.4±1.5 h/week; ST experience of 3.1±1.9 yrs and training volume of 2.5±1.4 h/week. 16 postmenopausal women (51.4±3.7 yrs; 56.7±8.3 kg; 161.7±4.9 cm): ET experience of 7.9±3.4 yrs and training volume of 4.1±1.2 h/week; ST experience of 3.1±1.9 yrs and training volume of 1.6±0.9 h/week
Who may be able to join
- Participants were required to meet the following criteria:
- Healthy adult females between 18 and 40 years old for eumerroheic and oral contraceptive groups or under 60 years old for postmenopausal women.
- Presenting with healthy iron parameters (serum ferritin >20μg/l, haemoglobin >115 μg/l and transferrin saturation >16%).
- Performing endurance training between 5 and 12 h per week (study I) or experienced in resistance training performing at least 1 h session two times per week during a minimum of a year (study II).
Who cannot join
- The exclusion criteria included:
- Irregular menstrual cycles.
- Any existing disease and/or metabolic or hormonal disorder.
- Any musculoskeletal injury in the last six months prior to the beginning of the project.
- Any surgery interventions (e.g. ovariectomy) or other medical conditions that would be exacerbated by an eccentric resistance exercise protocol.
- Regular use of medication or dietary supplements that could affect the results (e.g. nonsteroidal anti-inflammatory drugs).
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- Taking medication that alters vascular function (e.g. tricyclic antidepressants, α-blockers, β-blockers, etc.).
- Pregnancies in the year preceding.
- Smoking.
Trial locations and contacts
1 site in 1 country.
Spain1 site
Laboratorio de Fisiología Del Esfuerzo. Facultad de Ciencias de La Actividad Física Y Del Deporte. Universidad Politécnica de Madrid.
Madrid, 28040
About this study
This project is an observational controlled randomized counterbalance study. One hundred and three physically active and healthy women were selected to participate in the IronFEMME Study, of which 57 were eumenorrheic, 30 were oral contraceptive users (OCP) and 16 were postmenopausal women. The project consisted on two sections carrying out at the same time: Iron metabolism (Study I) and Muscle damage (Study II). For the study I, the exercise protocol consisted on an interval running test (8 bouts of 3 min at 85% of the maximal aerobic speed), whereas the study II protocol was based on an eccentric-based resistance exercise protocol (10 sets of 10 repetitions of plate-loaded barbell parallel back squats at 60% of their 1RM with 2 min of rest between sets). In both studies, eumenorrheic participants were evaluated at three specific moments of the menstrual cycle: Early-follicular phase, late-follicular phase and mid-luteal phase; OCP performed the trial at two moments: Withdrawal phase and active pill phase. Lastly, postmenopausal women were tested only once, since their hormonal status does not fluctuate. The three-step method was used to verify the menstrual cycle phase: calendar counting, blood analyses confirmation and urine-based ovulation kits. Blood samples were obtained to measure sexual hormones (e.g., 17β-Estradiol, Progesterone), iron metabolism parameters (e.g., Hepcidin, Iron, Ferritin, Transferrin) and muscle damage related markers (e.g., Creatine Kinase, Myoglobin, Lactate Dehydrogenase).
- Conditions
- Iron-deficiency, Inflammation, Exercise Induced Iron-deficiency, Exercise Induced Muscle Damage, Regulation of Sex Hormones on Hepcidin, Regulation of Sex Hormones on Muscle Damage
- Being tested
- Interval running protocol / eccentric-based resistance exercise protocol
- Keywords
- Hepcidin, Iron deficiency, Menstrual cycle, Muscle damage, Creatine kinase, Female, Exercise, Estrogen, Progesterone, Oral contraceptives, Postmenopausal women
- Medical subject headings
- Anemia, Iron-Deficiency, Inflammation, Iron Metabolism Disorders, Iron Deficiencies, Motor Activity
What participants receive
What participants receive. In a randomised study each person is assigned to one group by chance.
Interval running protocol / eccentric-based resistance exercise protocolProcedure or surgery
| Study group | Role | What they receive |
|---|---|---|
| Eumenorrheic women | The project consisted on two sections carrying out at the same time: Iron physiology (Study I) and Muscle damage (Study II).
For the study I, the exercise protocol consisted on an interval running test. 5 min warm-up at 60% of the vVO2peak followed by 8 bouts of 3 min at 85% of the vVO2peak with 90 secs recovery at 30% of the vVO2peak between bouts. Finally, a 5 min cool down was performed at 30% of the vVO2peak.
The study II protocol was based on an eccentric-based resistance exercise protocol consisted on 10 sets of 10 reps of plate-loaded parallel back squats at 60% of their previously calculated 1RM with 2 mins recoveries between sets.
In both studies, eumenorrheic participants were evaluated at three specific moments of the menstrual cycle: Early-follicular phase (EFP), late-follicular phase (LFP) and mid-luteal phase (MLP); Receives: Procedure: Interval running protocol / eccentric-based resistance exercise protocol | |
| Oral contraceptive users | The project consisted on two sections carrying out at the same time: Iron physiology (Study I) and Muscle damage (Study II).
For the study I, the exercise protocol consisted on an interval running test. 5 min warm-up at 60% of the vVO2peak followed by 8 bouts of 3 min at 85% of the vVO2peak with 90 secs recovery at 30% of the vVO2peak between bouts. Finally, a 5 min cool down was performed at 30% of the vVO2peak.
The study II protocol was based on an eccentric-based resistance exercise protocol consisted on 10 sets of 10 reps of plate-loaded parallel back squats at 60% of their previously calculated 1RM with 2 mins recoveries between sets.
Oral contraceptive users performed the trial at two moments: Withdrawal phase (WP) and active pill phase (APP). Receives: Procedure: Interval running protocol / eccentric-based resistance exercise protocol | |
| Postmenopausal women | he project consisted on two sections carrying out at the same time: Iron physiology (Study I) and Muscle damage (Study II).
For the study I, the exercise protocol consisted on an interval running test. 5 min warm-up at 60% of the vVO2peak followed by 8 bouts of 3 min at 85% of the vVO2peak with 90 secs recovery at 30% of the vVO2peak between bouts. Finally, a 5 min cool down was performed at 30% of the vVO2peak.
The study II protocol was based on an eccentric-based resistance exercise protocol consisted on 10 sets of 10 reps of plate-loaded parallel back squats at 60% of their previously calculated 1RM with 2 mins recoveries between sets.
Postmenopausal women were tested only once, since their hormonal status does not fluctuate. Receives: Procedure: Interval running protocol / eccentric-based resistance exercise protocol |
What is being measured
The primary outcome is the main question the study is designed to answer.
Hepcidin
Hepcidin is a protein that in humans is encoded by the HAMP gene. Hepcidin is a key regulator of the entry of iron into the circulation in mammals
Measured: pre-exercise
Hepcidin
Hepcidin is a protein that in humans is encoded by the HAMP gene. Hepcidin is a key regulator of the entry of iron into the circulation in mammals
Measured: 0 hours post-exercise
Hepcidin
Hepcidin is a protein that in humans is encoded by the HAMP gene. Hepcidin is a key regulator of the entry of iron into the circulation in mammals
Measured: 3 hours post-exercise
Hepcidin
Hepcidin is a protein that in humans is encoded by the HAMP gene. Hepcidin is a key regulator of the entry of iron into the circulation in mammals
Measured: 24 hours post-exercise
Creatine kinase
It is an enzyme that helps regulate the concentration of adenosine triphosphate within a cell. To do so, creatine kinase catalyzes the movement of a phosphate group from ATP to creatine, forming phosphocreatine. This molecules stores the phosphate group in a stable form, acting as an energy reservoir in cells.
Measured: pre-exercise
Creatine kinase
It is an enzyme that helps regulate the concentration of adenosine triphosphate within a cell. To do so, creatine kinase catalyzes the movement of a phosphate group from ATP to creatine, forming phosphocreatine. This molecules stores the phosphate group in a stable form, acting as an energy reservoir in cells.
Measured: 2 hours post-exercise
Creatine kinase
It is an enzyme that helps regulate the concentration of adenosine triphosphate within a cell. To do so, creatine kinase catalyzes the movement of a phosphate group from ATP to creatine, forming phosphocreatine. This molecules stores the phosphate group in a stable form, acting as an energy reservoir in cells.
Measured: 24 hours post-exercise
Creatine kinase
It is an enzyme that helps regulate the concentration of adenosine triphosphate within a cell. To do so, creatine kinase catalyzes the movement of a phosphate group from ATP to creatine, forming phosphocreatine. This molecules stores the phosphate group in a stable form, acting as an energy reservoir in cells.
Measured: 48 hours post-exercise
Secondary outcomes (34)
Iron
Measured: pre-exercise
Iron
Measured: 0 hours post-exercise
Iron
Measured: 3 hours post-exercise
Iron
Measured: 24 hours post-exercise
Transferrin
Measured: pre-exercise
Transferrin
Measured: 0 hours post-exercise
Transferrin
Measured: 3 hours post-exercise
Transferrin
Measured: 24 hours post-exercise
Ferritin
Measured: pre-exercise
Ferritin
Measured: 0 hours post-exercise
Ferritin
Measured: 3 hours post-exercise
Ferritin
Measured: 24 hours post-exercise
Mioglobin
Measured: pre-exercise
Mioglobin
Measured: 2 hours post-exercise
Mioglobin
Measured: 24 hours post-exercise
Mioglobin
Measured: 48 hours post-exercise
LDH
Lactate deshidrogenase
Measured: pre-exercise
LDH
Lactate deshidrogenase
Measured: 2 hours post-exercise
LDH
Lactate deshidrogenase
Measured: 24 hours post-exercise
LDH
Lactate deshidrogenase
Measured: 48 hours post-exercise
TNF-alfa
Measured: pre-exercise
TNF-alfa
Measured: 2 hours post-exercise
TNF-alfa
Measured: 24 hours post-exercise
TNF-alfa
Measured: 48 hours post-exercise
Interleukin-6
Measured: pre-exercise
Interleukin-6
Measured: 0 hours post-exercise
Interleukin-6
Measured: 2 hours post-exercise
Interleukin-6
Measured: 24 hours post-exercise
Interleukin-6
Measured: 48 hours post-exercise
CRP
C-reactive protein
Measured: pre-exercise
CRP
C-reactive protein
Measured: 0 hours post-exercise
CRP
C-reactive protein
Measured: 2 hours post-exercise
CRP
C-reactive protein
Measured: 24 hours post-exercise
CRP
C-reactive protein
Measured: 48 hours post-exercise
Timeline
From the start of the study to its completion; the mark is today. Estimated dates are the sponsor’s plan.
- First posted7 Jul 2020
- Study start1 Jan 2017
- Primary completion31 Mar 2019
- Study completion1 Jun 2020
Resources
- Official record on ClinicalTrials.govThe registry entry this page is built from, with its full change history.
- Publications (17)Papers the sponsor has linked to this study.
- Other trials for Iron-deficiencyStudies of the same condition, open and under way.
- What the trial phases meanHow a treatment moves from first tests in people to approval.
More details
For doctors and researchers: how the study is designed, who runs it, and its registry IDs.
Study design
- Study type
- Observational
- Observational model
- Cohort
- Time perspective
- Cross-sectional
- Enrolment
- 103 participants (actual)
- Target duration
- 2 Months
Sponsor and investigators
- Lead sponsor
- Universidad Politecnica de Madrid
- Sponsor type
- University, hospital or other
- Collaborators
- Ministerio de Economía y Competitividad, Spain
- Responsible party
- Principal investigator, ANA BELEN PEINADO LOZANO, Universidad Politecnica de Madrid
- Investigators
- Ana Belén Peinado, Study director, LFE Research Group. Universidad Politécnica de Madrid
IDs, dates and publications
- ClinicalTrials.gov ID
- NCT04458662
- Sponsor's protocol ID
- DEP2016-75387-P
- Acronym
- IronFEMME
- Status last verified
- Jul 2020
- First submitted
- 29 Jun 2020
- Last updated
- 7 Jul 2020
- Results posted
- No
- Data monitoring committee
- No
- FDA-regulated drug
- No
- FDA-regulated device
- No
- Individual data sharing
- No
Publications
Lehtihet M, Bonde Y, Beckman L, Berinder K, Hoybye C, Rudling M, Sloan JH, Konrad RJ, Angelin B. Circulating Hepcidin-25 Is Reduced by Endogenous Estrogen in Humans. PLoS One. 2016 Feb 11;11(2):e0148802. doi: 10.1371/journal.pone.0148802. eCollection 2016.
PubMed 26866603Hou Y, Zhang S, Wang L, Li J, Qu G, He J, Rong H, Ji H, Liu S. Estrogen regulates iron homeostasis through governing hepatic hepcidin expression via an estrogen response element. Gene. 2012 Dec 15;511(2):398-403. doi: 10.1016/j.gene.2012.09.060. Epub 2012 Oct 3.
PubMed 23041085Ikeda Y, Tajima S, Izawa-Ishizawa Y, Kihira Y, Ishizawa K, Tomita S, Tsuchiya K, Tamaki T. Estrogen regulates hepcidin expression via GPR30-BMP6-dependent signaling in hepatocytes. PLoS One. 2012;7(7):e40465. doi: 10.1371/journal.pone.0040465. Epub 2012 Jul 11.
PubMed 22792339Li X, Rhee DK, Malhotra R, Mayeur C, Hurst LA, Ager E, Shelton G, Kramer Y, McCulloh D, Keefe D, Bloch KD, Bloch DB, Peterson RT. Progesterone receptor membrane component-1 regulates hepcidin biosynthesis. J Clin Invest. 2016 Jan;126(1):389-401. doi: 10.1172/JCI83831. Epub 2015 Dec 14.
PubMed 26657863Yang Q, Jian J, Katz S, Abramson SB, Huang X. 17beta-Estradiol inhibits iron hormone hepcidin through an estrogen responsive element half-site. Endocrinology. 2012 Jul;153(7):3170-8. doi: 10.1210/en.2011-2045. Epub 2012 Apr 25.
PubMed 22535765Thompson B, Almarjawi A, Sculley D, Janse de Jonge X. The Effect of the Menstrual Cycle and Oral Contraceptives on Acute Responses and Chronic Adaptations to Resistance Training: A Systematic Review of the Literature. Sports Med. 2020 Jan;50(1):171-185. doi: 10.1007/s40279-019-01219-1.
PubMed 31677121McClung JP. Iron status and the female athlete. J Trace Elem Med Biol. 2012 Jun;26(2-3):124-6. doi: 10.1016/j.jtemb.2012.03.006. Epub 2012 May 7.
PubMed 22572041Kendall B, Eston R. Exercise-induced muscle damage and the potential protective role of estrogen. Sports Med. 2002;32(2):103-23. doi: 10.2165/00007256-200232020-00003.
PubMed 11817996Tiidus PM, Lowe DA, Brown M. Estrogen replacement and skeletal muscle: mechanisms and population health. J Appl Physiol (1985). 2013 Sep 1;115(5):569-78. doi: 10.1152/japplphysiol.00629.2013. Epub 2013 Jul 18.
PubMed 23869062Sim M, Dawson B, Landers G, Swinkels DW, Tjalsma H, Yeap BB, Trinder D, Peeling P. Oral contraception does not alter typical post-exercise interleukin-6 and hepcidin levels in females. J Sci Med Sport. 2015 Jan;18(1):8-12. doi: 10.1016/j.jsams.2013.11.008. Epub 2013 Nov 28.
PubMed 24373771Sipaviciene S, Daniuseviciute L, Kliziene I, Kamandulis S, Skurvydas A. Effects of estrogen fluctuation during the menstrual cycle on the response to stretch-shortening exercise in females. Biomed Res Int. 2013;2013:243572. doi: 10.1155/2013/243572. Epub 2013 Sep 12.
PubMed 24151587Janse DE Jonge X, Thompson B, Han A. Methodological Recommendations for Menstrual Cycle Research in Sports and Exercise. Med Sci Sports Exerc. 2019 Dec;51(12):2610-2617. doi: 10.1249/MSS.0000000000002073.
PubMed 31246715Romero-Parra N, Barba-Moreno L, Rael B, Alfaro-Magallanes VM, Cupeiro R, Diaz AE, Calderon FJ, Peinado AB. Influence of the Menstrual Cycle on Blood Markers of Muscle Damage and Inflammation Following Eccentric Exercise. Int J Environ Res Public Health. 2020 Mar 2;17(5):1618. doi: 10.3390/ijerph17051618.
PubMed 32131554Guisado-Cuadrado I, Romero-Parra N, Bell M, Elliott-Sale KJ, Sale C, Klentrou P, Peinado AB. Resistance and interval running exercise exert differential short-term post-exercise bone metabolic marker responses in postmenopausal. Bone. 2026 Aug;209:117910. doi: 10.1016/j.bone.2026.117910. Epub 2026 Apr 27.
PubMed 42055105Guisado-Cuadrado I, Romero-Parra N, Cupeiro R, Elliott-Sale KJ, Sale C, Peinado AB. Effect of eccentric-based resistance exercise on bone (re)modelling markers across the menstrual cycle and oral contraceptive cycle. Eur J Appl Physiol. 2025 May;125(5):1463-1473. doi: 10.1007/s00421-024-05693-y. Epub 2024 Dec 30.
PubMed 39738864