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Research background hyperprolactinemia (Hyperprolacfinemia, HPRL) is a gynecological frequently-occurring disease, hypothalamic - pituitary - sexual axis dysfunction caused by an endocrine disease, can women amenorrhea, menstrual disorders, infertility, reproductive function decline to elevated serum prolactin in the blood as the main clinical manifestations. In recent years, with the blood radioimmunoassay check the application and popularization of the (blood radioimmunoassay, RIA), the clinical diagnosis of the disease rate significantly improved pharmaceutical workers more and more attention to this disease. HPRL not only the breast lactation, but also has the role of inhibition of fertility and destruction of gonadal function. Excessive PRL form a feedback on the hypothalamus and inhibit the release of gonadotropin-releasing hormone. Inhibition of pituitary follicle-stimulating hormone (FSH), luteinizing hormone (LH) secretion, reduce the secretion or loss of rhythm of secretion, not secrete FSH midcycle LH peak. In addition, HPRL inhibit the function of ovarian granulosa cells to produce progesterone (P), also inhibited the temptation of the LH receptor, inhibition of ovarian response to FSH and LH response capability, resulting in a low estrogen state. When high blood PRL, light FSH secretion, resulting in ovarian follicular hypoplasia in the number of granule cells become less after ovulation, the secretion of progesterone reduced to luteal function is not perfect, severe cases, ovulation ability can completely lost. The ergot alkaloids Bromocriptine (Bromocriptine) is the drug of choice for domestic treatment HPRL, mostly imported. However, 12% of patients can not tolerate the side effects of bromocriptine a therapeutically effective amount. The drug's main side effects are gastrointestinal reactions, the dose is large may have dizziness, orthostatic hypotension, headache, drowsiness and constipation reactions, patients are often difficult to insist on taking. Chinese medicine safety, side effects, costs are not advantages HPRL's application in the treatment of many years of basic and clinical research, and attention gradually. The growing body of literature has reported the positive role played by the traditional Chinese medicine in the treatment of HPRL. Suppression the milk Tiaojing particles (yiru tiaojing granule, YTG) is our hospital pharmacy unit to Chinese medicine theory, kidney-oriented basis HPRL the liver depression as the subject of traditional Chinese medicine pathogenesis, as well as modern pathogenesis, on the basis of a large number of animal experiments, according to TCM The dialectical and Li-fa Prescription granules made of modern pharmaceutical technology, mainly used in the treatment of hyperprolactinemia. Clinical studies have shown that: This product HPRL therapeutically effective rate of 94.7%, especially satisfactory clinical outcome of liver depression and kidney deficiency HPRL. Objective To investigate the YTG quality standards and pharmacodynamic evaluation, mainly for the following experiment: a thin layer chromatography to identify the main ingredient: visits YTG Curculigo, white peony root, licorice TLC identification. 2: Determination the YTG the main ingredient Curculigoside and Radix Determination. 3 acute toxicity test: To observe the acute toxicity of the mice YTG. Pharmacodynamic evaluation: research YTG control effect of of hyperprolactinemia experimental rats and ovariectomized rats. Method 1. Thin-layer chromatography (TLC method) of the the YTG in the Curculigoside, white peony root identify. Determination of 2.YTG main ingredient 2.1 YTG Curculigoside Determination of chromatographic conditions: mobile phase: 0.1% phosphoric acid: acetonitrile = 79:21; detection wavelength at 283nm; flow rate: 1mL/min; Column: USA Dumas diamond column (250mm × 4.6mm, 5μm); column temperature: 40 ℃. Under this condition, the linear range investigated, the negative control test, the precision of the real test, the recovery test, stability test YTG sample measurement. 2.2 YTG Determination of paeoniflorin mobile phase: acetonitrile: 0.1% phosphoric acid = 14:86; detection at 220 nm; flow rate: of 0.8mL/min; Column: American Dima Diamond column (250mm × 4.6mm, 5μm) ; Column temperature: room temperature. Under this condition, the linear range investigated, the negative control test, the precision of the real test, the recovery test, stability test YTG sample measurement. Female mice a single 3.YTG the Acute Toxicity Test ig acute toxicity preliminary experiments, observe the general condition and death situation, no death, and you can not find the median lethal dose days maximum concentration and maximum volume dose orally, were observed for 14 consecutive days, and to measure its maximum tolerated dose. Efficacy studies 4.1 YTG HPRL rat model of experimental model 4.YTG HPRL model and ovary removal. Dopamine receptor blocker hydrochloride metoclopramide injection by intraperitoneal administration caused rats HPRL model, observed suppression milk Tiaojing particles, in the low-dose group, Bromocriptine Mesylate Tablets positive control group, HPRL rat model group, normal control group serum prolactin (PRL), luteinizing hormone (LH) and progesterone (P), estradiol (E2), testosterone (T), follicle-stimulating hormone (FSH ) hormone levels as well as the spleen, ovaries organ coefficient. Efficacy and mechanism of the 4.2 suppression the milk Tiaojing particles to treat premature ovarian failure (POF). The ovariectomy method to create a model of premature ovarian failure, observed of suppression the milk Tiaojing particles ovariectomized rat organ index, estradiol levels. Results thin-layer chromatography can identify Curculigo, white peony, licorice, and negative without interference. 2.YTG main ingredient Determination of the concentration of the reference solution (μg / mL) the ordinate x peak area of ??the vertical coordinate Y as linear regression, was standard regression equation: Y = 11656X 4540.8 R2 = 1.0000 shows that fairy Mao glycosides injection volume showed a good linear relationship between the sample concentration and its peak area in the range of 0.023 ~ 3.0μg; the paeoniflorin standard regression equation: Y = 13898X 14 978, R2 = 0.9997 shows that the injection volume of paeoniflorin glycosides 0.0553- 7.080μg range of sample concentration and peak area showed a good linear relationship. The chromatographic method used without interference from the other components, precision, recovery, and good stability, the linear range can meet the needs of the measurement. Suppression the milk Tiaojing particles maximum dose mice 64.4g/kg/d, equivalent adult daily oral doses of 199.2 times, no mice died during the observation period. The four pharmacodynamic study 4.1 YTG rat model of experimental HPRL YTG, in low doses can be significantly reduced HPRL model serum prolactin (PRL) (P lt; 0.001) level; organ coefficient shows YTG able to maintain a normal quality of the organ, and the maintenance of the normal physiological functions of the various organs. 4.2 YTG YTG increase the efficacy of the treatment of premature ovarian failure (POF) ovariectomized rat uterus and adrenal weight, to prevent uterine and adrenal atrophy; ovariectomized rat vagina increased keratinized epithelial cells, and enhance the activity of estrogen, improve estradiol levels in the blood. Conclusion 1. Thin-layer chromatography exclusive, easy operation, good reproducibility, and can be used for the identification of the YTG. 2 using the HPLC method is not subject to interference from other components, precision, recovery, good stability, the linear range can meet the needs of the measurement. 3.YTG on female acute toxicity in mice show that: the drug safe and no adverse reactions. Based on the maximum tolerated dose test products pharmacology, pharmacodynamics study provides a basis. 4.YTG can improve the situation of disorder HPRL the model rat serum hormone levels, abnormal elevation of PRL, P, T decreased the effect of reduced secretion of FSH increase YTG has a good treatment of experimental hyperprolactinemia. Has a role to promote the secretion of estrogen and anti-POF the YTG ovariectomized rats.
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