Unexpected effect of the vehicle (grain ethanol) of homeopathic FSH on the in vitro survival and development of isolated ovine preantral follicles

The aims of this study were to investigate the effects of medium replacement system (experiment I) and of FSH presentations (homeopathic – FSH 6cH and allopathic FSH – rFSH; experiment II) on the in vitro development, hormone production and gene expression of isolated ovine preantral follicles cultu...

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Published in:	Microscopy research and technique Vol. 80; no. 4; pp. 406 - 418
Main Authors:	Lima, Lartiza F., Rocha, Rebeca M. P., Duarte, Ana Beatriz G., Brito, Ivina R., Silva, Gerlane M., Rodrigues, Giovanna Q., Nunes‐Pinheiro, Diana C. S., Sales, Antônia D., Moura, Arlindo A., Wheeler, Matthew B., Rodrigues, Ana Paula R., Campello, Cláudio C., Figueiredo, José Ricardo
Format:	Journal Article
Language:	English
Published:	United States Wiley Subscription Services, Inc 01-04-2017
Subjects:	Androgens Animals Apoptosis - genetics Caspase 3 - analysis Cell Proliferation - drug effects Connexin 43 - analysis Connexins - analysis Culture DNA Fragmentation Estradiol - biosynthesis Ethanol Ethanol - chemistry Ethanol - pharmacology Ethyl alcohol Experiments Female Follicle Stimulating Hormone - pharmacology Gap Junction alpha-4 Protein Gene expression Grains Homeopathy hormone Hormones - biosynthesis Hormones - pharmacology in vitro culture In vitro testing Organ Culture Techniques - methods Ovarian Follicle - drug effects Ovarian Follicle - growth & development ovarian follicles Progesterone - biosynthesis Recombinant Recombinant Proteins - pharmacology Sex hormones Sheep Vehicles ovarian follicles ethanol homeopathy in vitro culture hormone
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Summary:	The aims of this study were to investigate the effects of medium replacement system (experiment I) and of FSH presentations (homeopathic – FSH 6cH and allopathic FSH – rFSH; experiment II) on the in vitro development, hormone production and gene expression of isolated ovine preantral follicles cultured for 6 days. In experiment I, secondary follicles were cultured in the α‐MEM+ supplemented with FSH 6cH (0.05 fg/ml) or recombinant bovine FSH (100 ng/ml) without/with daily medium addition. The homeopathic FSH treatments with/without medium addition improved (p < .05) follicular development compared to rFSH100 treatment without addition. FSH 6cH with addition showed the highest (p < .05) estradiol production. To verify whether the effects of homeopathic FSH were not due to its vehicle, experiment II was performed. The α‐MEM+ was supplemented or not with alcohol (0.2% grain ethanol, v/v), FSH 6cH or rFSH100 with daily medium addition. Surprisingly, we found that all treatments improved follicular development compared to the α‐MEM+ (p < .05). Moreover, homeopathic FSH was similar to the other treatments including its vehicle. In conclusion, its vehicle (ethanol) causes the effect of homeopathic FSH on in vitro development of isolated ovine preantral follicles. Ethanol improved development of isolated ovine preantral follicles compared to the cultured control (α‐MEM+) after 6 days of culture. The homeopathic FSH was similar to its vehicle on the development of isolated preantral follicles after culture.
Bibliography:	REVIEW EDITOR: Dr. Paolo Bianchini This work was carried out at the Laboratory of Manipulation of Oocytes and Ovarian Preantral Follicles (LAMOFOPA), Faculty of Veterinary Medicine of Ceara' State University, Fortaleza, CE, Brazil, and supported by CNPq. Laritza Ferreira de Lima is a recipient of a grant from CAPES Brazil. In addition, Ana Paula Ribeiro Rodrigues and José Ricardo de Figueiredo are recipients of a grant from CNPq Brazil. Funding Information ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	1059-910X 1097-0029
DOI:	10.1002/jemt.22810