RESUMEN
The uterine tetanic contraction and uterine artery blood flow reduction are possible reasons for primary dysmenorrhea (PD). In the present study, we aimed to evaluate the uterine relaxant effect and the influence on uterine artery blood velocity of Ge-Gen Decoction (GGD), a well-known Chinese herbal formula. In female ICR mice, uterine contraction was induced by oxytocin exposure following estradiol benzoate pretreatment, and the uterine artery blood velocity was detected by Doppler ultrasound. Histopathological examination of the uterine tissue samples were performed by H&E staining. Ex vivo studies demonstrated that oxytocin, posterior pituitary, or acetylcholine induced contractions in isolated mouse uterus. GGD inhibited both spontaneous and stimulated contractions. In vivo study demonstrated that GGD significantly reduced oxytocin-induced writhing responses with a maximal inhibition of 87%. Further study demonstrated that GGD normalized oxytocin-induced abnormalities of prostaglandins F2 alpha (PGF2α) and Ca(2+) in mice. In addition, injection of oxytocin induced a decrease in uterine artery blood flow velocity. Pretreatment with GGD reversed the oxytocin response on blood flow velocity. Histopathological examination showed pretreatment with GGD alleviated inflammation and edema in the uterus when compared with the model group. Both ex vivo and in vivo results indicated that GGD possessed a significant spasmolytic effect on uterine tetanic contraction as well as improvement on uterine artery blood velocity which may involve PGF2α and Ca(2+) signaling, suggesting that GGD may have a clinic potential in PD therapy.
Asunto(s)
Medicamentos Herbarios Chinos/administración & dosificación , Dismenorrea/tratamiento farmacológico , Oxitocina/efectos adversos , Contracción Uterina/efectos de los fármacos , Animales , Velocidad del Flujo Sanguíneo/efectos de los fármacos , Dismenorrea/fisiopatología , Femenino , Humanos , Ratones , Ratones Endogámicos ICR , Útero/irrigación sanguínea , Útero/efectos de los fármacos , Útero/fisiopatologíaRESUMEN
Ge-Gen Decoction (GGD) is a classical formula of traditional Chinese medicine. It is generally used for treating common cold, fever and influenza in China and South East Asia. In this study, a systematic method was established for the qualitative and quantitative analysis of the major constituents in GGD. For qualitative analysis, a method of liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (Q-TOF MS/MS) was developed for identification of multi-constituents. Based on the UV spectra, retention time and MS spectra, sixty compounds in GGD extract were identified or tentatively characterized by comparing with reference substances or literatures. According to the qualitative results, a new quantitative analysis method of GGD was established by HPLC-DAD. Fourteen representative compounds unequivocally identified were chosen as marker components which were derived from five herbs in GGD excluding Zingiberis Rhizoma Recens and Jujubae Fructus. The analytical method was validated through intra- and inter-day precision, repeatability and stability, and the R.S.D. was less than 3.18%, 4.48%, 3.36% and 3.54%, respectively. The LODs and the LOQs for the analytes were less than 1.06 and 3.12µgmL(-1), respectively. The overall recoveries ranged from 94.8% to 105.6%, with the R.S.D. ranging from 0.68% to 3.23%. Then the new method was applied to determine twelve batches of GGD commercial products of three dosage forms. The results indicated that the new approach was applicable in the routine analysis and quality control of GGD products. The study might provide a basis for quality control of GGD, and further study of GGD in vivo.