Shandong University of Traditional Chinese Medicine, First Clinical Medical College, Jinan, China.
Integrative Medicine Research Centre of Reproduction and Heredity, The Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Jinan, China.
Front Endocrinol (Lausanne). 2023 Jan 20;14:1045384. doi: 10.3389/fendo.2023.1045384. eCollection 2023.
To study the effect of Guilu Erxian ointment on the outcome of IVF-ET in older patients with poor ovarian response infertility of kidney-qi deficiency type, and to verify and analyze the mechanism of action of traditional Chinese medicine on improving older patients with poor ovarian response infertility of kidney-qi deficiency type from the perspective of metabolomics using targeted metabolomics technology, identify the related metabolic pathways, and provide metabolic biomarker basis and clinical treatment ideas for improving older patients with poor ovarian response infertility.
This study was a double-blind, randomized, placebo-controlled trial, and a total of 119 infertile patients who underwent IVF-ET at Shandong Center for Reproduction and Genetics of Integrated Traditional Chinese and Western Medicine were selected. Eighty older patients with infertility undergoing IVF were randomly divided into older treatment group and older placebo group, and another 39 young healthy women who underwent IVF-ET or ICSI due to male factors were selected as the normal control group. Flexible GnRH antagonist protocol was used for ovulation induction in all three groups, and Guilu Erxian ointment and placebo groups started taking Guilu Erxian ointment and placebo from the third day of menstruation until IVF surgery. And ultra-high performance liquid chromatography-triple quadrupole mass spectrometer (UHPLC-QTRAP MS) was used to detect metabolites in the three groups of samples.
Compared with the placebo group, the number of oocytes retrieved, 2PN fertilization, high-quality embryos, total number of available embryos and estrogen on HCG day were increased in the treatment group, and the differences were statistically significant ( > 0.05), but the clinical pregnancy rate of fresh embryos and frozen embryos were not statistically significant ( > 0.05). The results of targeted metabolomics analysis showed that follicular fluid in the treatment group clustered with the normal young group and deviated from the placebo group. A total of 55 significant differential metabolites were found in the follicular fluid of older patients with poor ovarian response of kidney-qi deficiency type and patients in the normal young group, after Guilu Erxian ointment intervention, Metabolites such as L-Aspartic acid, Glycine, L-Serine, Palmitoleic Acid, Palmitelaidic acid, L-Alanine, Gamma-Linolenic acid, Alpha-Linolenic Acid, and N-acetyltryptophan were down-regulated, mainly involving amino acid metabolism and fatty acid metabolism.
Guilu Erxian ointment can effectively improve the clinical symptoms and IVF outcomes of older patients with poor ovarian response of kidney-qi deficiency type. There were differences in follicular fluid metabolites between older patients with poor ovarian response of kidney-qi deficiency type and normal women. L-Aspartic acid, L-Alanine, Aminoadipic acid, L-Asparagine, L-Arginine, L-Serine, Gamma- Linolenic acid, Pentadecanoic acid and Alpha-Linolenic Acid are closely related to older patients with poor ovarian response due to deficiency of kidney-qi and may be inferred as biomarkers. The mechanism of Guilu Erxian ointment intervention may be mainly through amino acid metabolism and fatty acid metabolism regulation.
研究龟鹿二仙膏对肾虚型卵巢反应不良高龄患者体外受精-胚胎移植结局的影响，并从代谢组学角度运用靶向代谢组学技术，验证和分析中药改善肾虚型卵巢反应不良高龄患者的作用机制，寻找相关代谢途径，为改善肾虚型卵巢反应不良高龄患者提供代谢标志物基础和临床治疗思路。
本研究为双盲、随机、安慰剂对照试验，选取山东中医药大学附属医院生殖医学中心行体外受精-胚胎移植的肾虚型卵巢储备功能不良不孕患者 119 例，将行体外受精-胚胎移植的 80 例肾虚型卵巢反应不良高龄不孕患者随机分为高龄治疗组和高龄安慰剂组，另选取因男方因素行体外受精或卵胞浆内单精子注射的 39 例年轻健康女性作为正常对照组。三组均采用灵活 GnRH 拮抗剂方案进行促排卵，高龄治疗组和高龄安慰剂组均于月经第 3 天开始服用龟鹿二仙膏和安慰剂，直至体外受精手术。采用超高效液相色谱-三重四极杆串联质谱（UHPLC-QTRAP MS）检测三组样本中的代谢物。
与安慰剂组相比，治疗组获卵数、2PN 受精率、优质胚胎数、可利用胚胎总数、HCG 日雌激素水平均升高，差异有统计学意义（>0.05），但新鲜胚胎和冷冻胚胎临床妊娠率差异无统计学意义（>0.05）。靶向代谢组学分析结果显示，治疗组卵泡液与正常年轻组聚类，与安慰剂组分离。肾虚型卵巢反应不良高龄患者和正常年轻组患者的卵泡液中发现 55 个差异有统计学意义的差异代谢物，经龟鹿二仙膏干预后，L-天门冬氨酸、甘氨酸、L-丝氨酸、棕榈油酸、棕榈烯酸、丙氨酸、γ-亚麻酸、α-亚麻酸、N-乙酰色氨酸等代谢物下调，主要涉及氨基酸代谢和脂肪酸代谢。
龟鹿二仙膏可有效改善肾虚型卵巢反应不良高龄患者的临床症状和体外受精结局。肾虚型卵巢反应不良高龄患者与正常女性卵泡液代谢物存在差异，L-天门冬氨酸、丙氨酸、氨己酸、L-天门冬酰胺、L-精氨酸、L-丝氨酸、γ-亚麻酸、十五烷酸和α-亚麻酸与肾虚型卵巢反应不良高龄患者密切相关，可推测为生物标志物。龟鹿二仙膏干预的作用机制可能主要通过调节氨基酸代谢和脂肪酸代谢。
