粒線體功能調控顆粒細胞的類固醇荷爾蒙生成對女性生育力的影響

Mitochondrial Function in Human Granulosa Cell Steroidogenesis Modulate Female fertility

吳文瀚(Wen-Han Wu)

指導教授 ： 高淑慧

臺北醫學大學/醫學院/醫學科學研究所/碩士(2008年)

https://doi.org/10.6831/TMU.2008.00042

摘要

顆粒細胞的功能被認為對卵細胞及胚胎的品質有直接的功能。本論文主旨在探討顆粒細胞粒線體功能對荷爾蒙生成，卵細胞成熟率、受孕力及胚胎品質的影響。我們接受人工生殖療程之患者的顆粒細胞，依照病因區分四大群組，包括子宮內膜異位(endometriosis)、多囊性卵巢症候群(polycystic ovary syndrome；PCOS)、女性其他不孕因素及男性不孕因素等四種分類，並以男性不孕因素的顆粒細胞為對照組。首先，分析患者血清及濾泡液中相關荷爾蒙指數，並測定顆粒細胞存活率、粒線體數量、粒線體膜電位變化。繼而分析與卵細胞數量、胚胎品質(embryo quality)、卵細胞受精率(fertilization rate)及患者懷孕率(pregnancy rate)之相關性。研究結果顯示雌激素(estradiol；E2)於四種群組中，在子宮內膜異位群組之患者血清中的E2含量為對照組之患者的61%(p=0.001)，尤其於患有巧克力囊腫的婦女其E2含量為對照組之46%(p<0.001)。子宮內膜異位群組其卵細胞數目為對照組的65% (p=0.007)；顆粒細胞的細胞凋亡率較高於對照組15%(p=0.045)、降低的粒線體數量(對照組的68%，p=0.024)、下降的粒線體膜電位(對照組之71%，p=0.022)與降低受精率(對照組的93%，p=0.093)。同樣情況也發現在多囊性卵巢症候群的群組上，包括高於對照組20%的細胞凋亡率(p=0.037)、粒線體數量的不足(對照組之72%，p=0.04)及受精率不佳(對照組之90%，p=0.019)。我們進而分析與類固醇荷爾蒙有關的生成酵素steroidogenic acute regulatory (StAR)及3β-hydroxysteroid dehydrogenase (3β-HSD)，我們發現其蛋白含量與hormone level有關(p<0.001)。人類顆粒細胞的粒線體功能障礙，阻礙類固醇荷爾蒙生成，並進而影響卵細胞成熟率、卵細胞受精率以及造成卵細胞品質不良和懷孕能力下降。

關鍵字

粒線體 ； 顆粒細胞 ； 類固醇荷爾蒙生成

並列摘要

The purpose of this study is to evaluate mitochondrial function in human granulosa cells from infertile patients and examine the relationship between mitochondrial dysfunction and cause of infertility. The specific aims are to clarify the relationship between hormone levels, mitochondrial function, granulosa cells steroidogenesis, fertility capacity and embryo quality. The granulosa cells were obtained from the individuals undergoing IVF-ET treatment. The granulosa cells were classified into four groups: endometriosis, polycystic ovary syndrome (PCOS), other female factor infertility and male factor infertility. The concentration of estradiol (E2) and progesterone were analyzed by radioimmunoassay. Cell viability, mitochondrial mass and mitochondrial membrane potential were detected by flow cytometry with propidium iodide, nonyl acridine orange, and N,N,N',N'-tetramethylethylenediamine, respectively. In addition, the oocyte numbers, oocyte quality, fertilization rate, and pregnancy rate were also recorded. The results showed, the significantly lower serum E2 levels were found in the endometriosis group (61% of the control, p=0.001). The serum E2 level from patients with chocolate cyst was declined to 46% of the control (p<0.001). In emdometriosis group, the numbers of obtained oocyte was decreased to 65% of the control (p=0.007). The cell apoptosis was 15% higher than the control. (p=0.045). Reduced mitochondrial mass (68% of the control，p=0.024), declined mitochondrial membrane potential (71% of the control，p=0.022), and poor fertilization rate (93% of the control, p=0.093) were found in endometriosis group. All of these findings were also demonstrated in the PCOS group, including high apoptosis rate (over 20% of the control, p=0.037), lower mitochondrial mass (72% of the control, p=0.04), and reduced fertilization rate (90% of the control, p=0.019). Moreover, the steroidogenic enzymes, steroidogenic acute regulatory (StAR) and 3β-hydroxysteroid dehydrogenase (3β-HSD) were also measured by Western blot. The enzyme contents of StAR and 3β-HSD were correlated to E2 levels (p<0.001). In conclusion, mitochondrial dysfunction of the human granulosa cells may contribute to the global decline of steroidogenesis, decersed fertilization rate, oocyte maturation rate and oocyte quality, and finally jeopardize fertility capacity.

並列關鍵字

mitochondria ； granulosa cell ； steroidogenesis ； StAR ； 3b_HSD

參考文獻

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