Effect of White Kwao Extract (Pueraria mirifica) on in vitro Development and Implantation Rate of Mouse Embryo

Authors: Sansani Rungrattawatchai

Abstract:

The White Kwao (Pueraria mirifica), a potent phytoestrogenic medicinal plant, has long been use in Thailand as a traditional folkmedicine. However, no scientific information of the direct effect of White Kwao on the development of mammalian embryo was available. Therefore, the purpose of this study was to investigate the effect of White Kwao extract on the in vitro development and implantation rate of mouse embryos. This study was designed into two experiments. In the first experiment, the two-cell stage mouse embryos were collected from the oviduct of superovulated mature female mice, and randomly cultured in three different media, the M16, M16 supplemented with 0.52μg esthinylestradiol-17β, and M16 supplemented with 10 mg/ml White Kwao extract. The culture was incubated in CO2 incubator at 37 oC . After the embryos were cultivated, the developments of embryos were observed every 24 hours for 5 days. The development rate of embryos from the two-cell stage to blastocyst stage in the media was with White Kwao was significantly higher (p<0.05) than those of the control group (68.50% versus 43.50%) but did not differ from the positive control group (68.50% versus 57.66%). In the second experiment, hatched blastocysts, which obtained from three different media, were differently labeled the nuclei with two polynucleotide-specific fluorochromes, the propidium iodide (PI) and the bisbenzimide. The results showed that the number of trophectoderm cells in the blastocysts that cultivated in the media with White Kwao did not significantly differ from the control (80.00 versus 70 cells) and the positive control group (80.00 versus 112.50 cells). The average number of inner cell mass in the White Kwao treated group did not significantly differ from the control group (20.50 versus 16.00 cells) and the positive control group (20.50 versus 20.50 cells). The total cell number including the trophectoderm and the inner cell mass of the individual hatched blastocyst was evaluated. The cell number in the blastocysts obtained from the media with the White Kwao did not significantly differ from the control (94.25 + 9.50 versus 92.33 + 4.05) and the positive control group (94.25 + 9.50 versus 110.33 + 9.16). The results demonstrated that the White Kwao treatment group did have a stimulating effect on the in vitro development of mouse embryos. The exact mechanism that White Kwao stimulated mouse embryo development is not known. The suspect mechanism may in a manner similar to the mechanism that of estrogen stimulated the development of the mouse embryos. Futher studies are needed to transfer the blastocyst into the endometrium of pseudopreagnancy mice to evaluate the effect of White Kwao on implantation

Keywords: White Kwao (Pueraria mirifica), blastocyst.

Digital Object Identifier (DOI): doi.org/10.5281/zenodo.1061814

References:

[1] Luang Anusansoonton, "Textbook of Kwao Khruea," 1931, Chiang Mai. Upatipong Pressing.
[2] U.Radomsuk, "Effect of Peuraria Mirifica extracts on the Reproductive of American Cockroaches," 1992, Chiang Mai. Thesis for Master of Science. Chiang Mai University.
[3] Nawapol. (no date). "Root of Pueraria Mirifica (online)" Available:http://www.nawapol.com/Pueraria.htm.(2000,November 11).
[4] S. Chansakaow et al., "Identification of the deoxymiroesterol as the actual rejuvenating principle of Kwao Keur, Peuraria Mirifica," 2000, Washington DC. ACS Publication.
[5] J.L. Ingham et. Al., "Coumestans from the root of Peuraria Mirifica," 1988, Z. Naturforsch Ser C. Zurich.
[6] S. Swadipong, "Development of screening test for oestrogenic of extracts of some northern Thai plant and investigation of their effects on mammary gland development in mice," 1979, Thesis for Master of Science. Chiang Mai University.
[7] D. Songkaew, "Effect of White Gwow (Peuraria Mirifica Shaw et Suvat.)," 1983, Thesis for Master of Science. Chiang Mai University.
[8] P.G. Grootuis, et al., "Estrogen and the endometrium: lesson learned from gene expression profiling in rodent and human," 2007, Oxford. Oxford University Press.
[9] P. Bowman and A. McLaren, "Cleavage rate of mouse embryo in vivo and in vitro," 1970, OCLC New Jersey.
[10] G.M. Harlow and P. Quinn, "Development of preimplantation mouse embryo in vivo and in vitro," 1982, Australia.
[11] T. Ducibella and E. Anderson, "Cell shape and membrane changes in the eight-cell mouse: prerequisites for morphogenesis of the blastocyst," 1975, Elsevier Inc.
[12] B.C. Paria and S.K. Dey, "Preimplantation embryo development in vitro:cooperative interactions among embryos and role of growth factor," 1990, Proc. Natl. Acad. Sci. USA.
[13] R.B. Dickson and M.B. Lippman, "Estrogenic regulation of gene and polypeptide growth factor secretion in human breast carcinoma," 1987, Stanford University High Wire Press.
[14] D.J. Hill., "Growth factor and their cellular action," 1991, USA.
[15] E.M. Walter and M.B. Wheeler., "The effect of the phytoestrogen, daidzein on in vitro development of procine embryos," Avilable:http://porknet,Outreach.edu. 2002.(2002, August 23).
[16] P. Bhatnagar et al., "CSF-1 and mouse preimplantation development in vitro," 1995, UK. British Fertility Society Published.