A Bufadienolide-Enriched Fraction of Bryophyllum pinnatum Inhibits Human Myometrial Contractility In Vitro

Abstract

Bryophyllum pinnatum has been used since the 1970s to prevent premature labour, first in anthroposophic hospitals and, more recently, also in the main Swiss perinatal centres. However, it is not known which compounds in B. pinnatum leaves contribute to the tocolytic effect. Here we studied the effects of a flavonoid-enriched fraction, the corresponding flavonoid aglycon mixture, a bufadienolide-enriched fraction, and B. pinnatum leaf press juice on human myometrial contractility in vitro. The strength (area under the curve and amplitude) and frequency of contractions were recorded using strips of human myometrium mounted in an organ bath system. Cell viability assays were performed with the human myometrium hTERT-C3 and PHM1 – 41 cell lines. Repeated addition of the flavonoid-enriched fraction, flavonoid aglycon mixture, bufadienolide-enriched fraction, or B. pinnatum leaf press juice led to a progressive decrease of contraction strength, without jeopardising the vitality of myometrium strips. The bufadienolide-enriched fraction was the most active, since 1 µg/mL of the bufadienolide-enriched fraction lowered the area under the curve to 40.1 ± 11.8% of the initial value, whereas 150 µg/mL of the flavonoid-enriched fraction, 6.2 µg/mL of the flavonoid aglycon mixture, and 10 mg/mL of the B. pinnatum leaf press juice were required to achieve comparable inhibition. A progressive increase of contraction frequency was observed, except in the case of the flavonoid aglycon mixture, which did not affect frequency. None of the test substances decreased myometrial cell viability, even at concentrations of 500 µg/mL of the flavonoid-enriched fraction, 40 µg/mL of the flavonoid aglycon mixture, 3.8 µg/mL of the bufadienolide-enriched fraction, and 75 mg/mL of the B. pinnatum leaf press juice, i.e., higher than those used in the myometrium experiments. Given the concentrations of flavonoids in the flavonoid-enriched fraction and B. pinnatum leaf press juice, and of bufadienolides in the bufadienolide-enriched fraction and B. pinnatum leaf press juice, it appears that bufadienolides may be mainly responsible for the relaxant effect.

Key words

Bryophyllum pinnatum - Crassulaceae - myometrium - flavonoids - bufadienolides - toxicity

Publication History

Received: 20 August 2018

Accepted after revision: 27 November 2018

Publication Date:
18 December 2018 (online)

• References

• 1 Kamboj A, Rathour A, Kaur M. Bufadienolides and their medicinal utility: a review. Int J Pharm Pharm Sci 2013; 5: 20-27
• 2 Biswas SK, Chowdhury A, Das J, Hosen SZ, Uddin R, Rahaman MS. Literature review on pharmacological potentials of Kalanchoe pinnata (Crassulaceae). Afr J Pharm Pharmacol 2011; 5: 1258-1262
• 3 Hamburger M, Potterat O, Fürer K, Simões-Wüst AP, von Mandach U. Bryophyllum pinnatum-reverse engineering of an anthroposophic herbal medicine. Nat Prod Commun 2017; 12: 1359-1364
• 4 Fürer K, Simões-Wüst AP, von Mandach U, Hamburger M, Potterat O. Bryophyllum pinnatum and related species used in anthroposophic medicine: constituents, pharmacological activities, and clinical efficacy. Planta Med 2016; 82: 930-941
• 5 Hassauer W, Schreiber K, von der Decken D. Ein neuer Weg in der tokolytischen Therapie. Erfahrungsheilkunde 1985; 34: 683-687
• 6 Fürer K, Simões-Wüst AP, Winkler A, Amsler N, Schnelle M, von Mandach U. Die Anwendung von Bryophyllum pinnatum-Präparaten in der Geburtshilfe und Gynäkologie – eine multizentrische prospektive Beobachtungsstudie. Complement Med Res 2015; 22: 231-236
• 7 Schenkel L, Simões-Wüst AP, Hösli I, von Mandach U. Drugs in pregnancy and lactation – medications used in Swiss obstetrics. Z Geburtshilfe Neonatol 2018; 222: 1-14
• 8 Betschart C, von Mandach U, Seifert B, Scheiner D, Perucchini D, Fink D, Geissbühler V. Randomized, double-blind placebo-controlled trial with Bryophyllum pinnatum versus placebo for the treatment of overactive bladder in postmenopausal women. Phytomedicine 2013; 20: 351-358
• 9 Lambrigger-Steiner C, Simões-Wüst AP, Kuck A, Fürer K, Hamburger M, von Mandach U. Sleep quality in pregnancy during treatment with Bryophyllum pinnatum: an observational study. Phytomedicine 2014; 21: 753-757
• 10 Plangger N, Rist L, Zimmermann R, von Mandach U. Intravenous tocolysis with Bryophyllum pinnatum is better tolerated than beta-agonist application. Eur J Obstet Gynecol Reprod Biol 2006; 124: 168-172
• 11 Simões-Wüst AP, Lapaire O, Hösli I, Wächter R, Fürer K, Schnelle M, Mennet-von Eiff M, Seifert B, von Mandach U. Two randomised clinical trials on the use of Bryophyllum pinnatum in pre-term labour – results after discontinuation. Complement Med Res 2018; 25: 269-273
• 12 Simões-Wüst AP, Al Hassani T, Müller-Hübenthal B, Pittl S, Kuck A, Meden H, Eberhard J, Decker M, Fürer K, von Mandach U. Sleep quality improves during treatment with Bryophyllum pinnatum: an observational study on cancer patients. Integr Cancer Ther 2015; 14: 452-459
• 13 Chang HH, Larson J, Blencowe H, Spong CY, Howson CP, Cairns-Smith S, Lackritz EM, Lee SK, Mason E, Serazin AC. Preventing preterm births: trends and potential reductions with current interventionsin 39 very high human development index countries. Lancet 2013; 381: 223
• 14 World Health Organization. Preterm birth. Available at (Accessed April 11, 2018): http://www.who.int/mediacentre/factsheets/fs363/en/
• 15 Romero R, Dey SK, Fisher SJ. Preterm labor: one syndrome, many causes. Science 2014; 345: 760-765
• 16 Schleußner E. The prevention, diagnosis and treatment of premature labor. Dtsch Ärztebl Int 2013; 110: 227
• 17 Hubinont C, Debiève F. Prevention of preterm labour: 2011 update on tocolysis. J Pregnancy 2011; 2011: 941057
• 18 ACOG: American college of Obstetricians and Gynecologists. Practice Bulletin No. 171: Management of preterm Labor. Obstet Gynecol 2016; 128: e155-e164
• 19 Gwehenberger B, Rist L, Huch R, von Mandach U. Effect of Bryophyllum pinnatum versus fenoterol on uterine contractility. Eur J Obstet Gynecol Reprod Biol 2004; 113: 164-171
• 20 Wächter R, Brenneisen R, Hamburger M, Mennet M, Schnelle M, Worel A, Simões-Wüst AP, von Mandach U. Leaf press juice from Bryophyllum pinnatum (Lamarck) Oken induces myometrial relaxation. Phytomedicine 2011; 19: 74-82
• 21 Simões-Wüst AP, Grãos M, Duarte C, Brenneisen R, Hamburger M, Mennet M, Ramos M, Schnelle M, Wächter R, Worel A. Juice of Bryophyllum pinnatum (Lam.) inhibits oxytocin-induced increase of the intracellular calcium concentration in human myometrial cells. Phytomedicine 2010; 17: 980-986
• 22 Fürer K, Raith M, Brenneisen R, Mennet M, Simões-Wüst AP, von Mandach U, Hamburger M, Potterat O. Two new flavonol glycosides and a metabolite profile of Bryophyllum pinnatum, a phytotherapeutic used in obstetrics and gynaecology. Planta Med 2013; 79: 1565-1571
• 23 Oufir M, Seiler C, Gerodetti M, Gerber J, Fürer K, Mennet-von Eiff M, Elsas SM, Brenneisen R, von Mandach U, Hamburger M. Quantification of bufadienolides in Bryophyllum pinnatum leaves and manufactured products by UHPLC-ESIMS/MS. Planta Med 2015; 81: 1190-1197
• 24 Vogel JP, Nardin JM, Dowswell T, West HM, Oladapo OT. Combination of tocolytic agents for inhibiting preterm labour. Cochrane Database Syst Rev 2014; (07) CD006169
• 25 Bachmann S, Betschart C, Gerber J, Fürer K, Mennet M, Hamburger M, Potterat O, von Mandach U, Simões-Wüst AP. Potential of Bryophyllum pinnatum as a detrusor relaxant: an in vitro exploratory study. Planta Med 2017; 83: 1274-1280
• 26 Cassidy A, Minihane AM. The role of metabolism (and the microbiome) in defining the clinical efficacy of dietary flavonoids. Am J Clin Nutr 2017; 105: 10-22
• 27 Braune A, Blaut M. Bacterial species involved in the conversion of dietary flavonoids in the human gut. Gut Microbes 2016; 7: 216-234
• 28 Kolodziejczyk-Czepas J, Stochmal A. Bufadienolides of Kalanchoe species: an overview of chemical structure, biological activity and prospects for pharmacological use. Phytochem Rev 2017; 16: 1155-1171
• 29 Fürer K, Eberli D, Betschart C, Brenneisen R, De Mieri M, Hamburger M, Mennet-von Eiff M, Potterat O, Schnelle M, Simões-Wüst AP, von Mandach U. Inhibition of porcine detrusor contractility by the flavonoid fraction of Bryophyllum pinnatum – a potential phytotherapeutic drug for the treatment of the overactive bladder syndrome. Phytomedicine 2015; 22: 158-164
• 30 Condon J, Yin S, Mayhew B, Word RA, Wright W, Shay J, Rainey WE. Telomerase immortalization of human myometrial cells. Biol Reprod 2002; 67: 506-514
• 31 Devost D, Zingg HH. Novel in vitro system for functional assessment of oxytocin action. Am J Physiol Endocrinol Metab 2007; 292: E1-E6
• 32 Ray SD, Fariss MW. Role of cellular energy status in tocopheryl hemisuccinate cytoprotection against ethyl methanesulfonate-induced toxicity. Arch Biochem Biophys 1994; 311: 180-190