An Extract of Pomegranate Fruit and Galangal Rhizome Increases the Numbers of Motile Sperm: A Prospective, Randomised, Controlled, Double-Blinded Trial

Double-Blinded Trial. PLoS ONE 9(10): e108532. doi:10.1371/journal.pone.0108532 An Extract of Pomegranate Fruit and Galangal Rhizome Increases the Numbers of Motile Sperm: A Prospective, Randomised, Controlled, Double-Blinded Trial Jens Fedder 0 Maja D. K. Fedder 0 Henrik B. Jakobsen 0 Ina Giversen 0 Lars P. Christensen 0 Erik T. Parner 0 Samuel Kim, University of Kansas Medical Center, United States of America 0 1 Laboratory of Reproductive Biology, Scientific Unit, Regional Hospital of Horsens, Horsens, Denmark , 2 Nerthus ApS, Lejre , Denmark , 3 Department of Chemical Engineering, Biotechnology and Environmental Technology, Faculty of Engineering, University of Southern Denmark , Odense, Denmark , 4 Department of Public Health, Section for Biostatistics, University of Aarhus , Aarhus , Denmark , 5 Centre of Andrology, Fertility Clinic, Department D, Odense University Hospital , Odense , Denmark Pomegranate fruit (Punica granatum) and galangal (Alpinia galanga) have separately been shown to stimulate spermatogenesis and to increase sperm counts and motility in rodents. Within traditional medicine, pomegranate fruit has long been used to increase fertility, however studies on the effect on spermatogenesis in humans have never been published. With this study we investigated whether oral intake of tablets containing standardised amounts of extract of pomegranate fruit and powder of greater galangal rhizome (Punalpin) would increase the total number of motile spermatozoa. The study was designed as a prospective, randomized, controlled, double-blinded trial. Enrolment was based on the mean total number of motile spermatozoa of two ejaculates. The participants delivered an ejaculate after 4-8 days of tablet intake and two ejaculates just before they stopped taking the tablets. Seventy adult men with a semen quality not meeting the standards for commercial application at Nordic Cryobank, but without azoospermia, were included in the study. Participants were randomized to take tablets containing extract of pomegranate fruit (standardised with respect to punicalagin A+B, punicalin and ellagic acid) and freeze-dried rhizome of greater galangal (standardised with respect to 19S19-acetoxychavicol acetate) or placebo on a daily basis for three months. Sixty-six participants completed the intervention (active treatment: n = 34; placebo: n = 32). After the intervention the total number of motile spermatozoa was increased in participants treated with plant extracts compared with the placebo group (p = 0.026). After three months of active treatment, the average total number of motile sperm increased by 62% (from 23.4 to 37.8 millions), while for the placebo group, the number of motile sperm increased by 20%. Sperm morphology was not affected by the treatment. Our findings may help subfertile men to gain an improved amount of motile ejaculated sperm by taking tablets containing preparations of pomegranate fruit extract and rhizome of greater galangal. Trial Registration: ClinicalTrials.gov NCT01357044 PLOS ONE | www.plosone.org - Data Availability: The authors confirm that all data underlying the findings are fully available without restriction. All relevant data are within the paper and its Supporting Information files. Funding: The study was funded by Centre for Science and Research of Alternative Treatment (VIFAB), who played no other role in the study. Grant number 1002282. VIFAB does not exist anymore. Nerthus ApS is owned by authors Ina Giversen (IG) and Henrik Byrial Jakobsen (HBJ). Nerthus ApS did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the author contributions section. Competing Interests: Two of the authors (IG and HBJ) are commercializing the product; however, their company, Nerthus APS, has not contributed financially to the study. The full names of the two filed patent applications are: Dry preparation of Alpinia galanga or Alpinia conchigera with high content of 19S-19acetoxychavicol acetate:(Application no. PCT/EP2014/061880); and Composition for enhancing semen quality in a male subject: (Application no. PCT/EP2014/ 061851). The product name of the food supplement: PunalpinH. There are no further patents, products in development, or marketed products to declare. This does not alter the authors adherence to all the PLOS ONE policies on sharing data and materials, as detailed online in the guide for authors. In the Western world, a decline in sperm quality during the last 40 years has been suggested [1,2], although the decrease may have been stabilized during the last decade [3]. Only 23% of Danish men have optimal semen quality and 15% are at high risk of needing fertility treatment in order to biologically father a child [3]. It is evident that any improvement in poor quality semen brings about an increased chance for obtaining pregnancy, also with regard to assisted reproductive technology. Intracytoplasmic sperm injection is often the only alternative offered to couples, where the male suffers from poor semen quality. This makes randomised controlled trials of supplements, drugs or diet advices, aimed to improve semen quality, important in order to create other solutions helpful for males with poor semen quality. Pomegranate (Punica granatum) and greater galangal (Alpinia galanga) belong to a limited group of well-known edible plants, which show promising effects in vivo on rodent sperm quality without causing undesirable side effects [47]. The pomegranate fruit consists of a leathery pericarp containing numerous seeds, each covered by a juicy, translucent aril. Both pericarp and aril juice contains hydrolysable tannins, with the pericarp being the far most concentrated source [8]. In the pericarp, the ellagitannins punicalagin A and B predominates, while punicalins, which are partial hydrolysis products of punicalagins [9], constitute a minor fraction. The ellagitannins are only found in trace amounts in the aril juice [8]. Instead, other hydrolysable tannins prevail in the juice together with another important group of polyphenols, the anthocyanins [10]. Ellagitannins and, to a lesser degree, anthocyanins possess pronounced antioxidative activity in vitro [8]. Juice from P. granatum fruit has been shown to stimulate spermatogenesis, to increase sperm motility, and to decrease the percentage of abnormal sperm in rats after seven weeks of gavage [4]. An increase in sperm motility and concentration together with a decrease in abnormal sperm rate was observed after 6 weeks of oral administration of pomegranate juice extract to rats [5]. Further, oral administration of pomegranate pericarp extract has been shown effective in reversing induced spermatogenic disruption in rats [6]. The ginger-like rhizome of A. galanga contains a wide variety of semivolatile and volatile compounds, the dominating substances being (...truncated)