补肾调经方对IVF-ET患者卵母细胞质量及卵泡液生殖激素的影响

Effect of Bushen Tiaojing Recipe on the Quality of the Oocytes and Reproductive Hormones in the Follicular Fluid in IVF-ET Patients

目的研究补肾调经方对体外受精-胚胎移植(in vitro fertilization-embryo transfer,IVFET)患者卵母细胞质量及与卵泡液生殖激素及颗粒细胞骨形态发生蛋白-15(bone morphogenetic protein-15,BMP-15)表达的影响。方法将60例符合纳入标准的接受IVF-ET治疗的不孕症患者,按照治疗顺序分为中药联合控制性超排卵组(治疗组,20例)和单纯控制性超排卵组(对照组,40例)。比较两组年龄、不孕年限、基础促卵泡刺激素(basal follicle-stimulating hormone,bFSH)水平、促性腺激素(gonadotropins,Gn)使用天数及使用量、皮下注射重组人绒毛膜促性腺激素(HCG)日血清中雌二醇(estradiol,E_2)、孕酮(progesterone,P)、促黄体生成素(luteotropic hormone,LH)水平、获卵数、受精率、胚胎数、优质胚胎率和.临床妊娠率。应用化学发光法检测卵泡液生殖激素FSH、LH、E_2、P、睾酮(testosterone,T)浓度。应用实时荧光定量PCR(real-time PCR)法和蛋白质印迹法(Western blot)测定成熟卵泡颗粒细胞BMP-15 mRNA和蛋白定量的表达。结果 60例患者妊娠32例,总妊娠率为53.3%。其中对照组40例,妊娠19例,妊娠率47.5%;治疗组20例,妊娠13例,妊娠率65.0%,两组比较,差异无统计学意义(P>0.05)。治疗组Gn用量低于对照组,优质胚胎率高于对照组,差异均有统计学意义(P<0.05)。而HCG注射日血清E_2、LH、P浓度,获卵数和受精率两组比较,差异无统计学意义(P>0.05)。与对照组比较,治疗组卵泡液FSH浓度均降低,LH浓度增高,妊娠患者卵泡液LH浓度亦明显高于未妊娠患者,差异均有统计学意义(P<0.05)。而卵泡液E_2、T、P浓度比较,差异无统计学意义(P>0.05)。治疗组颗粒细胞上BMP-15 mRNA和蛋白定量的表达较对照组均增高,妊娠患者亦高于未妊娠患者(P<0.05)。结论在IVF-ET过程中,补肾调经方可以提高卵母细胞质量,增加卵泡对外源性促性腺激素的敏感性。补肾调经方的这一作用与颗粒细胞上BMP-15 mRNA和蛋白定量的表达变化及卵泡液中FSH和LH的浓度变化有关。

Objective To explore the effect of Bushen Tiaojing Recipe （BTR） on the quality of o- ocytes, reproductive hormones, and the expression of bone morphogenetic protein-15 （BMP15） of in vitro fertilization-embryo transfer （IVF-ET） patients. Methods Sixty infertility patients who prepared for IVF-ET were assigned to two groups according to the treatment order, the treatment group [20 cases,treated with BTR ＋controlled ovarian hyperstimulation （COH）1 and the control group （treated with COH alone, 40 cases）. Age, the time limit for infertility, basal follicle-stimulating hormone （bFSH） concentra- tion, usage days and the dosage of gonadotropins （Gn）, serum levels of estradiol （E2）, luteotropic hor- mone （LH）, and progesterone （P） on the HCG injection day, the number of retrieved occytes, the fertili- zation rate, the number of embryos, the high quality embryo rate, and the clinical pregnancy rate were compared. Concentrations of follicle-stimulating hormone （FSH）, LH, E2, testosterone （T）, and P in the follicular fluid were detected via chemiluminescence microparticle immunoassay. The mRNA and protein expression of BMP-15 in mature granulosa cells was detected by real-time fluorescent PCR and Western blot. Results Thirty-two patients were pregnant and the total pregnancy rate was 53.3%. Of them, 19 were pregnant and the total pregnancy rate was 47.5% in the control group, while 20 were pregnant and the total pregnancy rate was 65.0% in the treatment group. But there was no statistical difference be- tween the two groups （P 〉0.05）. Compared with the control group, the Gn dosage was lower and the high quality embryo rate was higher in the treatment group, showing statistical difference （P 〈0.05）. There was no statistical difference in serum concentrations of E2, LH, or P on the HCG injection day, the num- ber of retrieved oocytes, or the fertilization rate （P 〉0.05）. Compared with the control group, FSH con- centrations in the follicular fluid were significantly lower and LH concentrations were significantly higher in the treatment group （P 〈 0.05）. The LH concentrations in the follicular fluid were significantly higher in pregnant patients than non-pregnant patients, showing statistical difference （P 〈0.05）. There was no sta- tistical difference in E2, T, or P concentrations （P 〉0.05）. The mRNA and protein expression of BMP-15 in granulosa cells was higher in the treatment group than in the control group （P 〈0.05）. It was also high- er in pregnant patients than non-pregnant patients, showing statistical difference （P 〈0.05）. Conclusion During the IVF-ET process, BTR could elevate the quality of oocytes, and increase the sensitivity of ovar- ian follicles to exogenous Gn, which was correlated with the mRNA and protein expression of BMP-15 in granulosa cells, and changing concentrations of FSH and LH.