二至天癸颗粒对肾气阴两虚型高龄IVF-ET患者卵泡液RBP-4及颗粒细胞AnnexinA5的影响

发布时间：2018-04-30 15:23

本文选题：高龄 + 肾气阴两虚　；参考：《山东中医药大学》2016年硕士论文

【摘要】：目的:在体外受精-胚胎移植(IVF-ET)过程中,观察补肾中药二至天癸颗粒对肾气阴两虚高龄不孕患者卵细胞质量的影响,并检测卵泡液中视黄醇结合蛋白4(RBP-4)以及卵巢颗粒细胞膜联蛋白A5(Annexin A5)表达,从细胞凋亡角度探讨补肾中药改善肾气阴两虚型高龄不孕患者卵细胞质量的可能作用机制。方法:1.选取中医辩证属肾气阴两虚型因输卵管因素接受IVF-ET助孕的高龄不孕患者66例,按照随机数字表法分为实验组33例(二至天癸颗粒联合西药),对照组33例(安慰剂联合西药)。两组患者分别给予二至天癸颗粒和安慰剂预处理3个月经周期。比较两组患者肾气阴两虚症候积分在治疗前后的变化情况,对比促性腺激素(gonadotropin,Gn)的用量以及用药天数,记录两组患者注射绒促性素(HCG)日的血清孕酮(P)、促黄体素(LH)水平,计算比较单个卵泡雌激素水平,统计比较两组患者的获卵数、优质卵率、受精率、优质胚胎率和临床受孕率。2.收集上述两组患者取卵日的卵泡液,应用酶联免疫吸附法(Enzyme Linked Immunosorbent Assay,Elisa)测定卵泡液中RBP-4的水平。3.收集颗粒细胞,应用蛋白免疫印迹(Western blot)技术测定Annexin A5的表达。结果:(1)实验组与对照组相比,其肾气阴两虚的症候得到明显的改善,差异有统计学意义(P0.05);(2)实验组患者Gn的用量以及天数少于对照组,孕酮(P)水平明显低于对照组,差异有统计学意义(P0.05);(3)HCG日单个卵泡雌二醇(E2/卵泡数)水平相比,实验组高于对照组,差异有统计学差异(P0.05);(4)与对照组相比,实验组的获卵数、优质卵率、受精率、优质胚胎率均较高,差异均有统计学意义(P0.05);(5)临床妊娠率方面,两组相比无统计学意义(P=0.1860.05),但实验组的妊娠率有明显升高的趋势;(6)实验组与对照组相比,卵泡液中RBP-4及颗粒细胞Annexin A5的表达水平均增加,差异有统计学意义(P0.05)。结论:1.中药二至天癸颗粒可明显改善高龄不孕患者的肾气阴两虚状态,联合Gn治疗不孕症可明显减少Gn的用量,缩短用药天数,在提高优质卵率、优质胚胎率方面疗效显著。2.二至天癸颗粒可能是通过上调卵泡液中RBP-4的含量,抑制颗粒细胞Annexin A5的凋亡来改善卵母细胞的成熟,从而提高卵细胞质量,有效改善高龄肾气阴两虚不孕患者的IVF治疗结局。
[Abstract]:Objective: to observe the effect of Erzhi Tiangui granule, a traditional Chinese medicine for tonifying the kidney, on the quality of oocyte in the elderly infertility patients with deficiency of kidney qi and yin during in vitro fertilization and embryo transfer. The expression of retinol binding protein 4 (RBP-4) and ovarian granular cell membrane binding protein (A5(Annexin A5) in follicular fluid was detected. From the point of view of cell apoptosis, the possible mechanism of improving the oocyte quality of senile infertility patients with deficiency of kidney qi and yin was discussed from the point of view of cell apoptosis. Method 1: 1. Sixty-six elderly infertile patients who received IVF-ET to help pregnancy due to the deficiency of kidney qi and yin in TCM were divided into experimental group (Erzhi Tiangui granule combined with western medicine) and control group (placebo combined with western medicine) according to random number table method. Two groups of patients were given 2-to-Tiangui granule and placebo pretreatment 3 menstrual cycle. To compare the changes of kidney qi and yin deficiency symptom score before and after treatment, to compare the dosage of gonadotropin (Gnn) and the days of administration of gonadotropin, and to record the serum progesterone, progesterone and luteinizing hormone (LH) levels of the two groups on the day of HCG injection. To calculate and compare the estrogen level of single follicle and compare the number of eggs, the rate of high quality eggs, the rate of fertilization, the rate of high quality embryo and the rate of clinical conception of the two groups. The follicular fluid of the above two groups was collected on the day of egg taking. The level of RBP-4 in the follicular fluid was measured by enzyme linked immunosorbent assay (enzyme-linked immunosorbent assay). Granulosa cells were collected and the expression of Annexin A 5 was detected by Western blotting. Results (1) compared with the control group, the symptoms of deficiency of kidney qi and yin were significantly improved in the experimental group (P 0.05) the dosage of Gn and the number of days in the experimental group were lower than those in the control group, and the level of progesterone was significantly lower than that in the control group. The level of estradiol E _ 2 / E _ 2 / follicle in the experimental group was higher than that in the control group, and the difference was statistically significant (P 0.05). Compared with the control group, the number of eggs obtained, the rate of good quality eggs, the rate of fertilization in the experimental group were higher than those in the control group. The pregnancy rate of the experimental group was significantly higher than that of the control group, but the pregnancy rate of the experimental group was higher than that of the control group, and there was no significant difference in the clinical pregnancy rate between the two groups. The expression of RBP-4 and Annexin A 5 in granulosa cells were increased in follicular fluid, and the difference was statistically significant (P 0.05). Conclusion 1. Erzhi Tiangui granule can obviously improve the state of deficiency of kidney qi and yin in elderly infertility patients. Combined with Gn treatment of infertility can obviously reduce the dosage of Gn, shorten the days of medication, and improve the rate of high quality eggs and high quality embryo rate. Erzhi Tiangui granule may improve the maturation of oocytes by upregulating the content of RBP-4 in follicular fluid and inhibiting the apoptosis of granulosa cells Annexin A5, thus improving the quality of oocytes and effectively improving the outcome of IVF treatment in elderly patients with deficiency of both qi and yin of kidney qi and yin.
【学位授予单位】：山东中医药大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R271.9

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