Clinical Research On The Effect Of Jingyue's Kidney-invigorating Pill On The Hormone Level, Follicular Development And Endometrium Of Rat Models With Ovulation Difficulties Due To Kidney Deficiency

ObjectiveOn the foundation of early researches, this randomized control study focuses on Jingyue's Kidney-invigorating Pill's effect on rat models' hormone levels and endometrial morphology who have ovulation difficulties due to kidney deficiency with the hope of giving objective evaluation of its curative effect and safety and providing necessary evaluation system and strict research material for the clinical research and new drug development in the prevention and treatment of ovulation difficulties by Chinese Medicine.MethodsThe study subjects were72female rats randomly divided into the Negative Control Group, the Model Group, Clomiphene Group and Kidney-invigorating Groups (high dose, medium dose and low dose) with12in each group. Modeling: Except the Negative Control Group's rats were given equal volume of intramuscular injection of normal saline, all the other groups' rats were given intramuscular injection of hydrocortisone injection5ml· kg-1· d-1. Conduct continuous modeling for14days. Administration:on the15th day of modeling, administer medicine at10ml/kg of the body weight of rats. The Kidney-invigorating High-, Medium-and Low-dose Groups are given Kidney-invigorating Pill solution at0.57g crude drug/ml,1.14g/ml,2.28g crude drug/ml. The Negative Control Group and the Model Group were given distilled water and the Clomiphene Group was given0.54mg/ml clomiphene citrate tablet liquid (clomiphene citrate) for15continuous days.1H after the last administration, etherize the orbital venous plexus for hemospasia. Kill rats with cervical dislocation method. Conduct anatomical sampling for the detection of corresponding indexes.Results1. Sexual Cycle:starting from the fifth day after modeling. Except the Negative Control Group, the rats in each group had prolonged and irregular vaginal exfoliate cell cycle. Since the11th day, the Negative Control Group had no obvious periodic change, while other groups showed irregularities in cycle and length, or even disappearance of the cycle.2. Weight:Compared with the Negative Control Group, Model Group rats on modeling D1, D7, D14, D29lost weight significantly (P<0.05). Compared with the Model Group, Clomiphene Group rats, after administration of the drug21and29days, lost weight significantly (P<0.05); Rats in Kidney-invigorating Pill Groups of different dose have no significant difference (P>0.05) compared with the Model Group in body weight at different time points.3. Feed consumption:Compared with the Negative Control Group, Model Group rats' feed consumption significantly reduced (P<0.05) on D1and D7after the administration. Compared with the Model Group, High-dose Group of Kidney-invigorating Pill rats'feed consumption significantly reduced (P <0.05) on D14, D21of administration. The Low-dose Group of Kidney-invigorating Pill rats'feed consumption significantly reduced (P <0.05) on D21of administration. The other groups'rats'feed consumption have no significant difference (P>0.05) at other time points.4. Water consumption:Compared with the Model Group, Rats in the Medium and High-dose Group of Kidney-invigorating Pill on D14significantly increased water consumption (P<0.05). The other groups' rats at other time points showed no significant difference (P>0.05) in drinking water consumption.5. Organ coefficient:Model Group rats' adrenal coefficients, compared with the Negative Control Group, significantly increased (P<0.05). Clomiphene Group rats' uterus coefficient and ovarian coefficient, compared with the Model Group, significantly decreased (P<0.05). Kidney-invigorating Pill High, Medium and Low-dose Groups, in terms of uterus and ovary organ coefficient, showed no significant difference (P>0.05) compared with Model Group.6. Levels of the hormone:Model Group rats'serum estuarial (E2) content, compared with the Negative Control Group, showed no significant difference (P>0.05). Each administration group rats'serum estuarial (E2) content, compared with that of the rats in the Model Group, showed no significant difference (P>0.05). Model Group rats'serum progesterone (P) levels, compared with Negative Control Group, decreased significantly (P<0.05). Kidney-invigorating Pill Low-dose Group rats'serum progesterone (P), compared with the Model Group, significantly increased (P<0.05). The remainder of each group rats'serum progesterone levels, compared with the Model Group, had no significant difference (P>0.05).7. Follicular Growth:Negative Control Group ovaries have visible follicles with normal number, large number of corpus luteum and without follicular cysts, hematoma or tumor. Model Group had visible ovarian follicles (primary and secondary follicles). Compared with the Negative Control Group, its corpus luteum obviously decreased in number and the atresic follicle increased significantly. The Clomiphene Group had visible growing follicles and follicular atresia which significantly increased compared with the Model Group. Kidney-invigorating Pill High-dose Group had visible ovarian follicle which increased significantly in number than the Model Group. Kidney-invigorating Pill Medium and Low-dose Groups, compared with Model Group, showed no significant difference in ovarian growing follicles, mature follicle, follicular atresia and number of corpus luteum.8. Endometrial morphologic changes:In the Negative Control Group, the uterine wall had clear layer structure, endometrium was thick, endometrial epithelium had single-layer columnar arrangement, submucosal lamina propria connective tissue had nonbranching uterine glands, glands was in large number and the myometrium and perimetrium had no obvious pathological changes. The Model Group, compared with the Negative Control Group, decreased obviously in the number of uterine glands and endometrial thickness. Clomiphene Group decreased obviously in the number of uterine gland and the endometrium thickness than the Model Group. Compared with the Model Group, the Kidney-invigorating Pill Groups of high dose, medium dose and low dose, in terms of endometrial thickness and uterine gland number, showed no significant difference.The study found that, compared with the Negative Control Group, the Model Group'ovarian growing follicle (primary follicle and secondary follicle) and corpus luteum number decreased obviously, atresic follicle number increased significantly and the number of uterine glands decreased obviously. Compared with the Model Group, Clomiphene Group's ovarian growing follicle and follicular atresia significantly increased in number and the number of uterine glands and endometrial thickness decreased obviously. Kidney-invigorating Pill High-dose Group had obviously increased ovarian growing follicles than the Model Group. The experimental results indicate Kidney-invigorating Pill with high dose can promote kidney-deficient SD rats' ovarian primordial follicle developing into primary follicle and secondary follicle.Conclusion1. The hydrocortisone established kidney-deficient rat models have ovulation inhibition performance. There's successful simulation of clinical types of ovulation difficulties due to deficiency of the kidney.2. King Yue Kidney-invigorating Pill can improve the follicular development in rat model of ovulation difficulties due to kidney deficiency and, enhance rats' serum P levels.3. Kidney-invigorating Pill had the function of promoting ovarian follicular development. With the adjustment of reproductive axis formed with kidney-menstruation-Chong and Ren Meridians-uterus. It helps to reinforce the kidney-essence, adjust qi and blood in Chong and Ren meridians and promote the follicular development and ovulation.