- 學位論文
淫羊藿苷和松果菊苷經皮貼片對恢復大鼠生殖系統功能的體內評估
In Vivo evaluation of Icariin and Echinacoside transdermal patches for rejuvenating reproductive system function in aged rats
摘要
生育能力下降是全球面臨的重大問題，而卵巢早衰和衰老是導致生育能力下降的兩大主要原因，也間接造成人口老齡化和醫療資源不足的壓力。本實驗室研究發現淫羊藿苷（Icariin，ICA）和松果菊苷（Echinacoside，ECH）中草藥提取物具有改善卵巢的功能，且提高卵巢顆粒細胞的生長速度和雌二醇的分泌。本論文之目的為利用HHC經皮貼片緩慢遞送ICA和ECH來治療卵巢功能衰竭之SD母鼠。實驗設計係從 25 週齡健康母鼠利用含ICA 和 ECH 之經皮貼片進行治療。每週進行兩次治療，持續 9 週，共進行 14 次治療。每兩週採集血液進行雌二醇測定，並以H&E染色評估ICA和ECH對老年母鼠生殖系統功能的影響。研究結果顯示：含ICA和ECH的經皮貼片可使老年母鼠血液中的雌二醇增加10～20%。ICA經皮貼片可直接刺激雌二醇的分泌，且促進卵巢顆粒細胞增殖，提高雌配子的產生。ECH經皮貼片則增加卵巢中的 LH 分泌，此與子宮頸上皮上鱗狀上皮的增殖有關，從而促進雌二醇的分泌和增加性慾。體內動物實驗結果與體外卵巢顆粒細胞培養結果有一致性，亦即ICA和ECH兩種草藥提取物可刺激雌二醇分泌，其中，低濃度 ICA 即可顯著增加卵巢顆粒細胞的生長。實驗中發現ICA和ECH可並無其他副作用或有組織增生的反應，顯示其安全性。綜合動物實驗結果： ICA 和 ECH 經皮貼片可緩慢釋放ICA 和 ECH達到改善老年SD母鼠生殖系統的功能。
並列摘要
The decline of fertility is a major global issue, premature ovarian failure and senescence are two major causes of fertility decline and is now causing some serious social issue, including population aging and medical pressure. Our previous studies showed that Icariin (ICA) and Echinacoside (ECH), two Chinese herb extracts could improve ovarian function, that was effective in improving the growth rate of and estrodiol secretion of ovary granulosa cells. In this present thesis, we evaluate the effects of ICA and ECH to treat ovarian failure in aged SD rats. We prepared two types of transdermal patches (ICA-HCC and ECH HCC transdermal patch) to treat female aged SD rats starting from 25 weeks old. The patch was applied on the dorsal site of aged rats twice a week for 9 weeks and with total of 14 treatments. Also, we repeated the experiments on different ages of rats acting as blind tests to assure the efficacy of these two herbal extracts on SD rats. Blood samples, morphology of reproductive system and H&E staining of the reproductive systems were harvested and prepared to determine the effectiveness of ICA and ECH on improving the functions of the reproductive system in aged SD rats. The preliminary results suggested that ICA and ECH improved the function of the female reproductive system with different mechanisms. In the rats’ ovary tissue slices, the 8-week-old rat have a total follicles number of 8, when treated with low concentration of ICA-HHC patches, the number increased to 6, and in another group treated with higher concentration of ICA-HHC patches the number further rises to 9. But in the group treated with ECH-HHC patches, both high and low concentration groups have a follicles number of 3, which are slightly lower than the control group. This shown ICA generates the development of follicles and ECH did not. Both ICA and ECH transdermal patches increased the estradiol secretion about 10~20%. Aged rats treated with ICA transdermal patch increased the secretion of estrodiol directly and resulted in increasing the proliferation of ovary granulosa cells to improve the production of female gametes. ICA transdermal patch could increase LH reaction in ovary and the proliferation of the squamous epithelium on the virginal epithelium on aged rats which lead to the secretion of estrodiol and increase of sexual desire. These results agree with our pervious in-vitro results, both ICA and ECH herbal extracts increase the secretion of estrodiol but only ICA significantly increased the growth of ovarian granulosa cells. In addition, ICA and ECH herbal extracts increased the secretion of estradiol about 4~5 folds and exhibited no obvious side effect and tissue hyperplasia on reproductive system, indicating the safety of these extracts. In summary, ICA and ECG transdermal patches revealed potential application to improve the function of reproductive system on aged SD rats.