Impact of homeopathic remedies on the expression of lineage differentiation genes: an in vitro approach using embryonic stem cells

Saras Jyoti; Simran Tandon

doi:10.1016/j.homp.2015.11.002

Homeopathy, Table of Contents

Homeopathy 2016; 105(02): 148-159
DOI: 10.1016/j.homp.2015.11.002

Original Paper

Impact of homeopathic remedies on the expression of lineage differentiation genes: an in vitro approach using embryonic stem cells

Saras Jyoti

¹Department of Biotechnology and Bioinformatics, Jaypee University of Information Technology, Waknaghat, 173234, India

,

Simran Tandon

²Amity University Uttar Pradesh, Sector 125, Noida, 201313, UP, India

› Author Affiliations

Abstract

Background: Well-documented studies of the potential effects and safety of homeopathic medicines in pregnancy are required. In this study, specific genes were studied which could serve as biomarkers for specification of three lineages to predict the safety of homeopathic remedies using mouse embryonic stem (ES) cells. Thus, the present work was to study the effects of homeopathic remedies taken during pregnancy using ES cells as the model.

Methods: Mouse ES cells were exposed to 30C potency of Nux Vomica and Sepia, which are homeopathic medicines prescribed for the management of pregnancy related symptoms. Cytotoxicity studies were done using a modified Embryonic Stem cell test (EST). The expression levels of key genes and proteins were analyzed using real time polymerase chain reaction and immunocytochemistry, respectively.

Results: Homeopathic treatment led to modulations in the expression of certain lineage specific genes but this difference was not significant with respect to solvent control and showed normal differentiation as demonstrated by the expression of α/β MHC and α-actinin proteins in the differentiated ES cells.

Conclusions: Our study for the first time has shown the feasibility of using ES cells in the developmental toxicity testing of remedies. The results suggest that they are not associated with developmental toxicity.

Keywords

Homeopathy - Embryonic stem cells - Differentiation - Pregnancy - Developmental toxicity - Abbreviations: - ES = embryonic stem - EB = embryoid body - Nes = nestin - ND200 = neurofilament 200 kDa - Afp = alpha fetoprotein - GAPDH = glyceraldehyde-3-phosphate dehydrogenase

Full Text

References

References
1 Deroukakis M. Selection of potencies by medical and non-medical homeopaths: a survey. Homeopathy 91 2002; 150-155.
2 Badell M.L., Ramin S.M., Smith J.A. Treatment options for nausea and vomiting during pregnancy. Pharmacotherapy 2006; 26: 1273-1287.
3 Jonas W.B., Kaptchuk T.J., Linde K. A critical overview of homeopathy. Ann Intern Med 2003; 138: 393-399.
4 Shang A., Huwiler-Müntener K., Nartey L. et al Are the clinical effects of homoeopathy placebo effects? Comparative study of placebo-controlled trials of homoeopathy and allopathy. Lancet 2005; 366 9487 726-732.
5 Weingartner O. What is the therapeutically active ingredient of homeopathic potencies?. Homeopathy 2003; 92: 145-151.
6 Saha S.K., Roy S., Khuda-Buksh A.R. Evidence in support of gene regulatory hypothesis: gene expression profiling manifests homeopathy effects more than placebo. Int J High Dilution Res 2013; 12: 162-167.
7 Preethi K., Ellanghiyil S., Kuttan G., Kuttan R. Induction of apoptosis tumor cells by some potentiated homeopathic drugs: implications of mechanism of action. Integ Cancer Ther 2012; 11: 172-182.
8 Bellavite P., Marzotto M., Olioso D., Moratti M., Conforti A. High dilution effects revisited. 2. Pharmacodynamic mechanisms. Homeopathy 2014; 103: 22-43.
9 Bigagli E., Luceri C., Bernardini S., Dei A., Filippini A., Dolara P. Exploring the effects of homeopathic Apis mellifica preparations on human gene expression profiles. Homeopathy 2014; 103: 127-132.
10 Moskowitz R. Homoeopathic medicines for pregnancy and childbirth. 1992. North Atlantic Books; Berkely California.:
11 Catherine R. Coulter portraits of homoeopathic medicines. Psychophysical analysis of selected constitutional types Vol. 1 2002. hb publication.;
12 Festag M., Viertel B., Steinberg P., Sehner C. An in vitro embryotoxicity assay based on the disturbance of the differentiation of murine embryonic stem cells into endothelial cells. II. Testing of compounds. Toxicol In Vitro 2007; 21: 1631-1640.
13 Zur Nieden N.I., Ruf L.J., Kempka G., Hildebrand H., Ahr H.J. Molecular markers in embryonic stem cells. Toxicol In Vitro 2001; 15: 455-461.
14 Central Council for Research In Homeopathy, (An Autonomous Body of Deptt. of AYUSH, Ministry of Health & Family Welfare, Govt. of India) India. Homeopathy for healthy motherhood.
15 Arora S., Aggarwal A., Singla P., Jyoti S., Tandon S. Anti-proliferative effects of homeopathic medicines on human kidney, colon and breast cancer cells. Homeopathy 2013; 102: 274-282.
16 Hahnemann S. Organon of the Medical Art. 6th edn. 1843. Birdcage Books; Redmond, WA.:
17 Genschow E., Spielman H., Scholz G. et al The ECVAM. International validation study on in vitro embryotoxicity tests: result of the definitive phase and evaluation of prediction models. Altern Lab Anim 2002; 30: 157-176.
18 Romero A.C., Vilanova E., Sogorb M.A. Shortening and improving the embryonic stem cell test through the use of gene biomarkers of differentiation. J Toxicol 2011; 286034: 8.
19 Heuer J., Bremer S., Pohl I., Spielmann H. Development of an in vitro embryotoxicity test using murine embryonic stem cell cultures. Toxicol In Vitro 1993; 7: 551-556.
20 Hogberg H.T., Kinsner-Ovaskainen A., Coecke S., Hartung T., Bal-Price A.K. mRNA Expression is a relevant tool to identify developmental neurotoxicants using an in vitro approach. Toxicol Sci 2010; 113: 95-115.
21 Kattman S.J., Huber T.L., Keller G.M. Multipotent Flk-1+ cardiovascular progenitor cells give rise to the cardiomyocyte, endothelial, and vascular smooth muscle lineages. Dev Cell 2006; 11: 723-732.
22 Shalaby F., Ho J., Stanford W.L. et al A requirement for Flk1 in primitive and definitive hematopoiesis and vasculogenesis. Cell 1997; 89: 981-990.
23 Hoh J.F., Mcgrath P.A., Hale P.T. Electrophorectic analysis of multiple forms of rat cardiomyosin: effects of hypophysectomy and thyroxine replacement. J Mol Cell Cardiol 1978; 10: 1053-1076.
24 Lints T.J., Parsons L.M., Hartley L., Lyons I., Harvey R.P. Nkx-2.5: a novel murine homeobox gene expressed in early heart progenitor cells and their myogenic descendants. Development 1993; 119: 419-431.
25 Kwon G.S., Fraser S.T., Eakin G.S. et al Tg (Afp-GFP) expression marks primitive endoderm lineages during mouse development. Dev Dyn 2006; 235: 2549-2558.
26 Podgornyi O.V. Differentiation and behavior of neural stem cells tissue culture and on transplantation into the rat brain. [in Russian], Author's Abstract of Doctoral Thesis in Biological Sciences, Moscow 2006.
27 Neilsen A.C. Survey backs homeopathy benefits. Business Standard. September 4, 2007 http://www.business-standard.com/india/news/a-c-nielsen-survey-backs-homeopathy-benefits/295891/ “Alternative Medicine/Alternative Medical Market,” Frost and Sullivan Ltd. Report #E874, London1986.
28 Alpert N.R., Mulieri L.A. Increased myothermal economy of isometric force generation in compensated cardiac hypertrophy induced by pulmonary artery constriction in the rabbit. Circ Res 1982; 50: 491-500.
29 Komuro I., Izumo S. Csx: a murine homeobox-containing gene specifically expressed in the developing heart. Proc Natl Acad Sci 1993; 90: 8145-8149.
30 Lyons I., Parsons L.M., Hartley L. et al Myogenic and morphogenetic defects in the heart tubes of murine embryos lacking the homeobox gene Nkx2-5. Genes Dev 1995; 9: 1654-1666.
31 Brock D.J., Bolton A.E., Monaghan J.M. Prenatal diagnosis of anencephaly through maternal serum-alphafetoprotein measurement. Lancet 1973; 2: 293-294.
32 Leek A.E., Ruoss C.F., Kitau M.J., Chard T. Raised AFP in maternal serum with anencephalic pregnancy. Lancet 1973; 2: 385-386.
33 Frenkel M., Mishra B.M., Sen S. et al Cytotoxic effects of ultra-diluted remedies on breast cancer cells. Int J Oncol 2010; 36: 395-403.