Combined Red Clover isoflavones and probiotics potently reduce menopausal vasomotor symptoms

RESEARCH ARTICLE Combined Red Clover isoflavones and probiotics potently reduce menopausal vasomotor symptoms Max Norman Tandrup Lambert1, Anne Cathrine Thorup1, Esben Søvsø Szoscka Hansen2, Per Bendix Jeppesen1* 1 Department of Endocrinology and Internal Medicine, Aarhus University Hospital, Aarhus, Denmark, 2 MR Research Centre, Aarhus University Hospital, Skejby, Denmark a1111111111 a1111111111 a1111111111 a1111111111 a1111111111 * Abstract Background OPEN ACCESS Citation: Lambert MNT, Thorup AC, Hansen ESS, Jeppesen PB (2017) Combined Red Clover isoflavones and probiotics potently reduce menopausal vasomotor symptoms. PLoS ONE 12(6): e0176590. https://doi.org/10.1371/journal. pone.0176590 Editor: Tiffany L. Weir, Colorado State University, UNITED STATES Received: November 14, 2016 Accepted: April 9, 2017 Published: June 7, 2017 Copyright: © 2017 Lambert et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Data Availability Statement: All data are uploaded in the public repository Figshare with the relevant DOI number: 10.6084/m9.figshare.4983368. Funding: This study was funded by Styrelsen for Forskning og Innovation and Future Food Innovation (FFI), a public non-profit organization supported by the European Union and the Region of Central Jutland Denmark. This study was also supported in part by Agro Food Park, Skejby, Denmark- The European Fond for Regional Development, Denmark and The Danish Ministry Natural estrogen decline leads to vasomotor symptoms (VMS). Hormone therapy alleviates symptoms but increases cancer risk. Effective treatments against VMS with minimal cancer risks are needed. We investigate the effects of a highly bioavailable aglycone rich Red Clover isoflavone treatment to alleviate existing menopausal VMS, assessed for the first time by 24hour ambulatory skin conductance (SC) Methods and results We conducted a parallel, double blind, randomised control trial of 62 peri-menopausal women aged 40–65, reporting  5 hot flushes/day and follicle stimulating hormone 35 IU/L. Participants received either twice daily treatment with bioavailable RC extract (RCE), providing 34 mg/d isoflavones and probiotics, or masked placebo formulation for 12 weeks. The primary outcome was change in daily hot flush frequency (HFF) from baseline to 12 weeks using 24hr SC. Secondary outcomes were change in SC determined hot flush intensity (HFI), self-reported HFF (rHFF) and hot flush severity (rHFS), blood pressure and plasma lipids. A significant decrease in 24hr HFF (P < 0.01) and HFI (P<0.05) was found when comparing change from baseline to 12 months of the RCE (-4.3 HF/24hr, CI -6.8 to -2.3; -12956 μS s-1, CI -20175 to -5737) with placebo (0.79 HF/24hr, CI -1.56 to 3.15; 515 μS s-1, CI -5465 to 6496). rHFF was also significantly reduced (P <0.05)in the RCE (-2.97 HFs/d, CI -4.77 to -1.17) group compared to placebo (0.036 HFs/d, CI -2.42 to 2.49). Other parameters were non-significant. RCE was well tolerated. Conclusion Results suggest that moderate doses of RCE were more effective and superior to placebo in reducing physiological and self-reported VMS. Findings support that objective physiological symptom assessment methods should be used together with self-report measures in future studies on menopausal VMS. PLOS ONE | https://doi.org/10.1371/journal.pone.0176590 June 7, 2017 1 / 16 Red Clover isoflavones reduce menopausal vasomotor symptoms for Research and Innovation, Copenhagen, Denmark. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Competing interests: ML, AT and PBJ are coinventors on the patent application “PCT/DK2013/ 050428” for the production of the Red Clover extract used in the present trial as required by US patent authorities. All rights have been assigned to the company Herrens Mark without any kind of compensation, where ML, AT and PBJ have forgone their rights as stipulated by terms and conditions of Aarhus University. Hence, the authors declare no financial or other interests. This does not alter our adherence to all the PLOS ONE policies on sharing data and materials. Abbreviations: BMI, body mass index; BP, blood pressure; CI, confidence interval; CVD, cardiovascular disease; ER, estrogen receptor; FSH, follicle stimulating hormone; GI, gastrointestinal; GCS, Greene climacteric scale; HDL, high density lipoprotein; HF, hot flushes; HFF, hot flush frequency; HFI, hot flush intensity; HPLC, high performance liquid chromatography; HT, hormone therapy; LDL, low density lipoprotein; RC, Red Clover; RCE, Red Clover extract; rHFF, self-reported hot flush frequency; rHFS, self-reported hot flush intensity; SC, skin conductance; SD, standard deviation; TC, total cholesterol; VLDL, very low density lipoprotein; VMS, vasomotor symptoms. Trial registration ClinicalTrials.gov NCT02028702 Introduction Menopause symptoms severely reduce the quality of life of women worldwide, up to 80% of women may experience symptoms and it is estimated that in 2030 the at risk groups of periand post- menopausal women will reach 1.2 billion globally [1]. The core symptoms are hot flushes (HF) and night sweats (NS), collectively referred to as vasomotor symptoms (VMS); sleep disturbance and other secondary symptoms often also present [2]. These symptoms are largely a consequence of natural endogenous estrogen decline and dysregulation during peri- and post- menopause; estrogen deficiency is further associated with increased risk of osteoporosis, cardiovascular disease (CVD) and negative changes to lipid profile [3–5]. Hormone therapy (HT) is the current gold standard treatment for VMS. However substantial evidence supports that therapy increases cancer risk in estrogen receptor (ER) α rich tissues (e.g. uterus, breast and ovaries) [6,7]. The IMS and Revised Global Consensus Guidelines recommend and agree that both estrogen only and combined treatments: increase cancer risk with longer duration of use, that physicians should undertake a case by case risk benefit assessment prior to treatment, that treatments be limited to 5 years, that cancer risk is affected by time from the start of menopause and initiation of HT treatment, that the lowest effective dose be given, that patients undertaking HT have annual monitoring and that HTs are unsuitable for use in breast cancer operated patients/survivors suffering from estrogen deficient VMS [8–10]. Currently, there are no recommendations regarding ovarian cancer risk, a recent meta-analysis of 52 studies has shown equivalent relative risk increases of 1.43 and 1.37 for both estrogen only and combined therapies used for < 5 years respectively [11]. Considering that the median total VMS duration is shown to be 7.4 years and that the current guidelines for (...truncated)