Studies Of Quality Standards And Pharmacodynamics On Yiru Tiaojing Granule

BackgroundHyperprolactinemia (HPRL) is a very common in gynecological diseases. HPRL is caused by dysfunction of the hypothalamus - pituitary - axis which can make female menopause infertility, menstrual disorders and reproductive dysfunction.The serum elevated prolactin in the blood is as main clinicalmanifestations. In recent years, with the application and popularization of blood radioimmunoassay (RIA),the clinical diagnosis of the disease was significantly increased,which caused a number of medical workers attached great importance to.Excessive amount of PRL made a feedback on hypothalamic, prevent the release of gonadotropin-releasing hormone, and inhibit the generation of pituitary follicle-stimulating hormone (FSH) and the secretion of luteinizing hormone (LH). In addition, HPRL can inhibit granulosa cells produce progesterone (P) role, and the temptation to suppress LH receptor, which inhibit the ovarian response capability on FSH and LH and result in low estrogen state.Currently, the first choice drug in domestic and foreign for treatment of HPRL is synthetic peptide ergot alkaloids bromocriptine. However,12% of patients can not tolerate its side effects under effective treatment dosage. The drug's side effects are mainly gastrointestinal reaction. Larger doses may have dizziness, orthostatic hypotension, headache, drowsiness and constipation and other reactions, so that patients are often difficult to insist on taking. Chinese medicines have the advantages of safety, fewer side effects, low cost and so on. The application in the treatment of HPRL has gone through years of basic and clinical research, and gradually be taken seriously. More and more literatures reported that Chinese medicines played a positive role in the treatment of HPRL.Yiru tiaojing granule(YTG) is a kind of puretraditional chinese medicine. The pharmaceutical preparation section and the gynecology and obstetrics experts in nanfang hospital have done a lot of practice and research on this drug。It is based on the Chinese medicine that HPRL is associated with kidney deficiency and Hyperactive Liver-fire. So the experts suggest one method Warming kidney and activating yang and Regulating Yin and Yang to cure HPRL. YTG is used to prevent and treathyperprolacfinemia in clinic. Clinical study demonstrates that its therapeutic efficiency ratio for HPRL amounts to 94.7%, especially for the kind of HPRL wih Kidney Deficiency and Liver Stasis.ObjectiveTo identify YTG; in order to the property of YTG, we studied its acute toxicity on female mice, studied the effects of YTG on experimental HPRL rats and on estrogen activity of ovariectomized rats.Method1. Identity the curculigo orchiodes and Paeonia lactiflora in YTG by Thin layer chromatographic method (TLC method)2. Detection the main components in YTG2.1 Studies on determination of the contents Curculigoside in YTG by HPLCThe mobile phase of Curculigoside was phosphate solution:acetonitrile (79: 21).Flow rate was 1 ml/min.The UV detective wavelength was 283nm. Chromatographic column Wasdikma diamond (250mm×4.6mm,5μm).Column temperature was 40℃. Under these conditions, we studied the linear range and did the negative control test, precision test, recovery test, stability test and the Sample determination of YTG.2.2 Studies on Determination of Paeoniflorin in YTG by HPLCThe mobile phase of Paeoniflorin was phosphate solution:acetonitrile (86: 14).Flow rate was 0.8 ml/min.The UV detective wavelength was 223nm. Chromatographic column was dikma diamond (250mm×4.6mm,5μm). Column temperature was room temperature.Under these conditions, the linear range was studied and the negative control test,precision test, recovery test, stability test and Sample determination of YTG were done.3.□The acute toxic reaction of YTG in female miceThe death rate and general states of the mice was used to evaluate the pre-experiment acute toxicity induced by YTG for 14 days. If mice were not dead, the median lethal dose (LD50) can not calculated. The maximum tolerated dose of YTGwas used to evaluate by administration of maximum concentration and maximum volume in a day.4. Pharmacodynamic action of YTG4.1 Effects of YTG on the experimental HPRL RatsThe HPRL rat models were made by continuously injecting hydrochloride metoclopramide, dopamine receptor blocker through abdominal cavity. Then the serum levels of PRL, LH, P, T, E2 and FSH were detected among the high, middle, low dose groups of YTG, the positive group of bromocriptine mesylate Tablets, the HPRL rat model group and the normal control group. At the same time, the organ indexes were also observed.4.2 Effect of YTG on estrogen activity of ovariectomized ratsThe rat model of premature ovarian failure (POF) was made by cutting the bilateral ovaries of female rats. Then the effects including serum E2 level,estrogen activity and organs index of Yiru Tiaojing Granule on ovariectomized Rats were observed.Result1 Thin-layer chromatography can identify Curculigo, Paeoniflorin, licorice and has no negative interference.2 The analytic research of main components in YTGThe method of HPLC used would not be interfered by other components and it has good precisio,recovery,stability. The range of linearity could satisfy the determination.3 The maximal dosage of YTG was 64.4g/kg/d by intragastric administration, which was 199.2 times of daily oral dosage for adults.4 Pharmacodynamic action of YTG4.1 Effects of YTG on the experimental HPRL RatsThe high, middle and low dosages of YTG could significantly lower the PRL level of HPRL rat model (P<0.001). The indexes showed that YTG could maintain the normal weight of organs and the normal physiological functions.4.2 Effect of YTG on estrogen activity of ovariectomized ratsYTG could increase the weight of womb and adrenal glands of ovariectomized rats and prevent their atrophy. It could raise the number of vaginal keratinized epithelial cells and the estradiol level in blood serum; and also enhanced estrogen activity.Conclusions1. The TLC methods were simple with strong specificity and good reproducibility and could be used to the identification of YTG2. The HPCL method used was not interferred by other components, which has good linear range, precision, recovery and stability. It can meet the needs determination of YTG.3. The acute toxicity of YTG was extremely lower. The results suggested that the safety were relatively higher. The security of YTG is satisfactory. The maximal dosage got from the experiment can provide more evidences for the pharmacological and pharmacodynamics research.4. YTG could improve the disorder condition of serum hormone level of HPRL rat models, decrease their serum levels of PRL, P and T and increase the FSH level. So YTG had a good treatment effect on experimental HPRL rat models. YTG could promote the secretion of estrogen and have a role of anti-POF.