Background: In the current society, infertility related to age has become a social problem. The in vitro fertilization (IVF) success rate in women with poor ovarian response (POR) is very low. Dandelion extract T-1 (DE-T1) is an effective component of the extract from the leaves and stems of Taraxacum officinale, which is one of the medicines used in some patients with POR, but its molecular mechanism remains unclear.
Methods: Following IVF, ovarian granulosa cells (GCs) of sixty patients were extracted and divided into normal ovarian response (NOR) and POR groups. GCs were cultured in a dose-dependent and time-dependent manner with DE-T1, proliferation of GCs was determined by Cell Counting Kit-8 assay, and mRNA levels of insulin-like growth factor 1 receptor (IGF-1R), luteotropic hormone receptor (LHR), follicle-stimulating hormone receptor (FSHR), LHR, and CYP19A1 (aromatase) were determined by quantitative polymerase chain reaction. Progesterone and estradiol (E2) concentrations were determined by enzyme-linked immunosorbent assay.
Results: The cell viability gradually increased with the progressive increase in the DE-T1 concentration. Compared with the control group (without DE-T1), the mRNA expressions of FSHR, LHR, IGF-1R, and CYP19A1 were upregulated after the addition of DE-T1, especially in the 2.5% DE-T1 group (P < 0.01). The expression of IGF-1R was upregulated approximately 25 times (24.97 ± 4.02 times) in the POR group with 2.5% DE-T1. E2 and progesterone levels increased with the increasing DE-T1 concentration. There were highly significant differences in the E2 and progesterone secretion between the NOR and POR groups (P < 0.01).
Conclusion: DE-T1 may promote steroid hormone synthesis by promoting GC proliferation and upregulating GC receptor expression, thereby improving ovarian endocrine function.
蒲公英提取物对促进人卵巢颗粒细胞增殖及激素受体表达的研究摘要背景：研究蒲公英提取物（DE-T1）对颗粒细胞增殖、甾体激素合成及相关受体表达的影响。 方法: 收集行体外受精-胚胎移植（in vitro fertilization -embryo transfer，IVF-ET）卵巢低反应组（poor ovary response，POR）和卵巢正常反应组（normal ovary response，NOR）患者各30例的卵泡液，并提取颗粒细胞，原代培养24 h后分别在含DE-T1低（1.25%）、中（2.5%）、高（5%）剂量的培养液内培养孵育48~60h，采用CCK-8法检测颗粒细胞活性。荧光定量PCR方法检测颗粒细胞胰岛素样生长因子1受体（insulin-like growth factor-1 receptor，IGF1-R）、促卵泡激素受体（follicle-stimulating hormone receptor, FSHR）、黄体生成素受体（luteinizing hormone，LHR）和CYP19A1mRNA的表达。采用酶联免疫吸附法（ELISA）测定颗粒细胞培养液中孕酮（progesterone，P）、雌二醇（Estradiol，E2）浓度。 结果：1、在一定浓度范围内（0.625%~5%），颗粒细胞活性随蒲公英提取物剂量增加而增强，其中1.25%、2.5%、5%DE-T1组，细胞增殖活性均显著优于未添加组（P＜0.01）。2、DE-T1加药后与空白对照组相比，FSHR、IGF-1R、CYP-19A1表达均有剂量依赖性的增高趋势。尤其在2.5%DE-T1加药组低反应患者LHR、IGF-1R、CYP-19A1mRNA表达上调倍数，均显著高于正常反应组，其中低反应组IGF-1R表达上调近25倍（24.97±4.02）。3、DE-T1加药后，正常反应及低反应两组颗粒细胞培养基中，雌激素和孕激素分泌均随DE-T1加药浓度的增加而升高。其中NOR组当DE-T1浓度达到2.5%时其雌二醇分泌水平较不加药组相比，有显著差异（P＜0.05），5%浓度时雌激素分泌出现极显著差异（P＜0.01），同时（5%浓度DE-T1下）孕激素的分泌量水平也有显著差异（P＜0.05）；在2.5%及5%浓度下，POR组在2.5%及5%浓度下，细胞雌二醇、孕激素水平二醇较空白组的分泌量均有显著增高（分别为P＜0.01、P＜0.05）。 结论：蒲公英提取物DE-T1可以促进颗粒细胞的增殖，促进FSHR、LHR、IGF-1R、CYP-19A1 mRNA的表达，促进卵巢颗粒细胞类固醇激素的合成。.
Keywords: Dandelion Extracts; Follicle-Stimulating Hormone Receptor; Human Granulosa Cells; Insulin-Like Growth Factor 1 Receptor; Proliferation; Steroidogenesis.