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Traditional Chinese medicine Guizhi Fuling capsule used for therapy of dysmenorrhea via attenuating uterus contraction

Authors Sun, Lan; Liu, Lina; Zong, Shaobo; Wang, Zhengzhong; Zhou, Jun; Xu, Zhiliang; Ding, Gang; Xiao, Wei; Kou, Junping

Journal JOURNAL OF ETHNOPHARMACOLOGY , SEP 15 2016,:191

DOI 10.1016/j.jep.2016.06.042 PubMed 27340106

Abstract Ethnopharmacological relevance: Guizhi Fuling formula, a well-known Chinese herbal formula recorded in the Eastern Han Dynasty, is composed of Cinnamomum cassia (L)J.Presl (Cassia bark), Poria cocos (Schw.) Wolf (Poria), Paeonia suffruticosa andrews (Mouton Cortex), Paeonia lactiflora Pall (Herbaceous peony), and Amygdalus persica L.(Persicae Semen). It has clinical efficacy of activating blood circulation to dissipate blood stasis and is commonly used for the treatment of primary dysmenorrhea. However, its therapeutic mechanism has not been clearly elucidated. The aim of this study is to reveal molecular mechanisms of action using in vivo and in vitro experimental models.Material and methods: The ICR mouse uterine contraction was induced by oxytocin exposure following estradiol benzoate pretreatment. Mice were given GZFLC (0.54, 1.08 g/kg) by gavage. The levels of NO, PGF(2 alpha), and Ca2+ in uterine tissue were determined according to instructions. Cyclooxygenase-2 (COX-2) and oxytocin receptor (OTR) proteins in uterine tissue were assessed by Western Blot. Mouse isolated uterus strips were mounted in tissue organ baths containing Locke''s solution. The contractile responses were recorded with Power Lab recording system. The effect of GZFLC on spontaneous uterine contraction, and uterine contraction induced by oxytocin, PGF(2 alpha) was observed. Myometrial cells were exposed to oxytocin (5 U/L) to induce calcium release, and the effect of GZFLC and its components (PL, PGG, CA) on intracellular Ca2+ was analyzed with fluorometry imaging.Results: In vivo study demonstrated that GZFLC significantly reduced oxytocin-induced writhing responses with a maximal inhibition of 55%. It also decreased the levels of NO, PGF(2 alpha) and Ca2+ in oxytocin-induced mice uterine tissue. Moreover, Western blot analysis showed that COX-2 and OTR expressions in uterine tissue of dysmenorrhea mice were significantly reduced. GZFLC inhibited spontaneous uterus contractions in a dose-dependent manner, and the IC50 value was 0.99 mg/ml. The IC50 values of GZFLC on PGF(2 alpha), oxytocin-induced contractions were 1.45 mg/ml, 3.53 mg/ml, respectively. Further in vitro studies indicated that GZFLC and its components (PL, PGG, CA) could restrain intracellular calcium levels in favour of uteri relaxation.Conclusions: Both in vivo and in vitro results indicated that GZFLC possessed a significant spasmolytic effect on uterine tetanic contraction. The present study provides in vivo and in vitro experimental evidence to support the use of GZFLC for the clinical treatment of primary dysmenorrheal (PD). (C) 2016 Elsevier Ireland Ltd. All rights reserved.

Species covered 肉桂 Cinnamomum cassia 芍药(原变种) Paeonia lactiflora 牡丹 Paeonia suffruticosa 桃 Amygdalus persica