补肾化痰方对PCOS模型大鼠卵巢形态及激素水平的影响

发布时间：2018-05-29 04:25

本文选题：PCOS + 补肾化痰方　；参考：《河南中医学院》2014年硕士论文

【摘要】：目的：本课题以传统中医学理论为基础，通过观察补肾化痰方对PCOS模型大鼠激素水平和卵巢组织形态的影响。揭示补肾化痰方降低大鼠促黄体生成素和雄激素水平，并改善大鼠卵巢组织形态的作用机理，为补肾化痰方治疗PCOS提供有效性科学依据。方法：该课题选用95只同种系雌性SPF清洁级21日龄大鼠，随机选择25只大鼠作为正常对照组，每日颈背部皮下注射0.2ml注射油剂，连续20d；其余70只大鼠作为病理模型组，每日颈背部皮下注射脱氢表雄酮（DHEA）6mg/100g体重+0.2ml注射油剂，连续20d。造模结束后，先行大鼠动情周期测定及OGTT实验，再从正常组和模型组中分别随机选取10只大鼠进行模型评估，选择造模成功大鼠进入下一步实验。有造模成功大鼠60只和正常组大鼠15只，将模型组大鼠60只随机分为模型对照组、二甲双胍组、中药低剂量组、中药高剂量组；每组各15只大鼠，正常组15只大鼠作为正常对照组。对各组大鼠连续灌胃4周后，观察其阴道涂片结果，对各组实验大鼠动情间期禁食不禁水12小时处理后称重、水合氯醛（10%）麻醉、腹主动脉取血、分离血清。对各组大鼠行断颈处死，卵巢组织用4%多聚甲醛固定，分离血清并测定激素水平，光镜下观察卵巢组织形态学的变化。结果：①实验过程中实验动物一般状态的变化：模型组大鼠于颈背部皮下注射脱氢表雄酮后出现活动量减少、饮食量增加、饮水量增加、尿量增加，体重增加不明显。以上变化随灌胃给药天数的增加而加剧，正常组大鼠体重逐渐增长、精神体征、活动反应如常。灌胃后，部分中药组大鼠体重下降，饮食、饮水较前减少。②本实验中大鼠血清水平的影响：模型组的大鼠血清促黄体生成素（LH）和雄激素（T）水平均高于正常组，差别有统计学意义（P0.05）；与模型组比较，二甲双胍组、中药高剂量组、低剂量治疗组血清LH和T水平均明显降低，差别有统计学意义(P0.05)；与二甲双胍组比较，高剂量中药组和低剂量中药治疗组血清LH和T水平均有下降，差别有统计学意义(P0.05)；中药高剂量组与低剂量组血清LH水平未见差别，结果无统计学意义（P0.05）；中药高剂量组大鼠血清T水平明显高于中药低剂量组，差别有统计学意义(P0.05)。③对实验大鼠卵巢组织形态学的影响：正常组卵巢组织：其可见各级卵泡，如初级卵母细胞、次级卵母细胞，生长卵泡细胞等，卵母细胞有放射冠，细胞结构排列紧密。可见数个颗粒细胞，黄体数有一定数目。模型组大鼠卵巢组织：其可见多个囊泡，其囊泡即是空卵泡。正常形态的各级细胞少见，甚至没有。见多个闭锁细胞，卵母细胞放射冠消失，颗粒细胞较少，细胞排列疏松，黄体数目较少。二甲双胍组大鼠卵巢组织：其可见极少数囊泡细胞，，各级卵泡细胞均在视野中可见且数量增多，可见到初级卵母细胞、次级卵母细胞、生长卵泡、成熟卵泡，即其成熟卵泡的卵泡膜也充分发育，内层细胞增大。卵母细胞可见放射冠、透明带。且看到颗粒细胞增多，细胞排列紧密；黄体数目增多。中药高剂量组大鼠卵巢组织：其可见少数囊泡细胞，视野中可见到少数初级卵母细胞、次级卵母细胞、生长卵泡以及成熟卵泡，颗粒细胞数目较模型组增多，可见一定数目黄体。中药低剂量组大鼠卵巢组织：其可见囊泡细胞数目减少，黄体数目稍有增加。可见少数生长卵泡细胞，以及少数闭锁细胞。各组大鼠镜下各级卵泡数目以及黄体数目比较：模型组大鼠的各级卵泡数及黄体数均低于正常组，差别有统计学意义（P0.05）。与模型组大鼠相比，二甲双胍组、高剂量治疗组、低剂量治疗组大鼠的各级卵泡数及黄体数均较多，差别有统计学意义（P0.05）。与二甲双胍组比较，高剂量治疗组、低剂量治疗组大鼠的各级卵泡数及黄体数数目有变化，差别有统计学意义（P0.05）。中药高剂量组与低剂量组大鼠的各级卵泡数及黄体数目间统计学数据无差异，无统计学意义（P0.05），统计结果虽然没有统计学意义，但卵泡数目和黄体数目有升高趋势。结论：①补肾化痰方能改善大鼠的生殖功能。②补肾化痰方能能直接作用于卵巢，并通过改善PCOS模型大鼠生殖内分泌的功能紊乱趋于稳定，从而影响大鼠卵巢的生殖功能，使得分泌的激素水平趋于正常。③本实验证实补肾化痰方能从根本上改善PCOS患者卵巢功能，进而能明显改善卵巢的组织形态结构，使其趋于正常卵巢功能。
[Abstract]:Objective: on the basis of traditional Chinese medicine theory, the effect of tonifying kidney and resolving phlegm prescription on hormone level and ovarian tissue morphology of PCOS model rats was observed, and the mechanism of tonifying kidney and eliminating phlegm prescription to reduce the level of luteinizing hormone and androgen in rats and to improve the morphology of ovarian tissue in rats was provided to provide the treatment of PCOS with the prescription of Tonifying the kidney and removing phlegm. Effective scientific basis.
Methods: 95 female SPF 21 day old rats were selected as the normal control group, and 25 rats were randomly selected as the normal control group. 0.2ml injection was injected subcutaneously on the back of the neck and 20d, and the other 70 rats were used as the pathological model group. The daily neck and back were subcutaneously injected with dehydroepiandrosterone (DHEA) 6mg/100g weight +0.2ml injection. After the end of 20d. model, 10 rats were randomly selected from the normal group and the model group. The model group and the model group were randomly selected for the model evaluation. The model rats were selected to enter the next experiment. There were 60 rats and 15 rats in the normal group, and 60 rats in the model group were randomly divided into the model control group and two a double group. Guanidine group, low dose group of traditional Chinese medicine, high dose group of traditional Chinese medicine, 15 rats in each group, and 15 rats in normal group as normal control group. After 4 weeks of continuous perfusion, the results of vaginal smear were observed in each group of rats. After 12 hours' treatment, the rats in each group were weighed, chloral hydrate (10%), abdominal aorta were taken blood, and blood was separated. The rats in each group were executed with broken neck, the ovarian tissue was fixed with 4% polyformaldehyde, the serum was separated and the level of hormone was measured. The morphological changes of the ovary were observed under light microscope.
Results: (1) changes in the general state of experimental animals during the experiment: after the hypodermic injection of dehydroepiandrosterone on the back of the model group, the amount of activity decreased, the amount of diet increased, the amount of drinking water increased, the amount of urine increased, and the increase of weight was not obvious. The above changes were aggravated with the increase in the number of days of administration of the stomach. The weight of the normal rats increased gradually. The influence of the serum level of rats in this experiment: the level of serum luteinizing hormone (LH) and androgen (T) in the rat model group was higher than that in the normal group, and the difference was statistically significant (P0.05); compared with the model group, metformin group, The serum levels of LH and T in the low dose group were significantly decreased, and the difference was statistically significant (P0.05). Compared with the metformin group, the level of serum LH and T decreased in the high dose group and the low dose treatment group, the difference was statistically significant (P0.05). There was no difference in the serum LH level between the high dose group and the low dose group. The results were not statistically significant (P0.05); the level of serum T in the high dose group of Chinese medicine was significantly higher than that of the low dose group of traditional Chinese medicine (P0.05). The effect of the difference was statistically significant (3) on the ovarian histomorphology of the experimental rats: the normal group of ovarian tissue: the follicles in the normal groups, such as primary oocytes, secondary oocytes, follicle cells, and so on, were found in the normal group. A number of granular cells and the number of corpus luteum have a certain number. The model group of rat ovarian tissue: it can be seen a number of vesicles, its vesicles are empty follicles. The normal form of cells at all levels are rare, even no. See multiple atresia cells, oocyte radiated crowns disappear, granulosa cells are fewer, cell rows are less. In the group of metformin group, the ovary tissue of metformin group: it can be seen in a few vesicles, and the follicle cells at all levels are visible and increased in the field of vision. It can be seen that the primary oocyte, secondary oocyte, growth follicle, mature follicle, that is, the follicular membrane of its mature follicle is fully developed, and the inner cell enlargement. The number of granulosa cells increased, the cells were arranged closely, and the number of corpus luteum increased. The high dose group of Chinese medicine showed a few vesicular cells, and a few primary oocytes, secondary oocytes, growth follicles and mature follicles in the field of vision, and the number of granulosa cells was more than that of the model group. The number of follicle cells and the number of corpus luteum increased slightly. The number of follicles at all levels and the number of corpus luteum were compared: the number of follicles and the number of corpus luteum at all levels in the model group were lower than that of normal rats. The difference was statistically significant (P0.05). Compared with the model group, the number of follicles and corpus luteum at all levels in the metformin group, the high dose treatment group and the low dose treatment group were more, the difference was statistically significant (P0.05). Compared with the metformin group, the number of follicles and the number of corpus luteum numbers in the low dose treatment group and the low dose treatment group were in the high dose treatment group. The difference was statistically significant (P0.05). There was no statistical difference between the number of follicles and the number of corpus luteum in the high dose group and the low dose group, and there was no statistical significance (P0.05). Although the statistical results were not statistically significant, the number of follicles and the number of corpus luteum increased.
Conclusion: (1) the prescription of Tonifying the kidney and removing phlegm can improve the reproductive function of the rat. (2) the prescription of Tonifying the kidney and removing phlegm can directly act on the ovary, and improve the functional disorder of the reproductive endocrine of the PCOS model rats, thus affecting the reproductive function of the rat ovary and the secretion of the hormone level to the normal. It can fundamentally improve ovarian function in PCOS patients, which can significantly improve the ovarian morphology and structure, and make it tend to normal ovarian function.
【学位授予单位】：河南中医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R285.5

【参考文献】