親生殖腺素釋放激素對於子宮內膜異位症患者中細胞外間質分子表現之影響

1. 親生殖腺素釋放激素對於子宮內膜異位症患者中細胞外間質分子表現之影響 子宮內膜異位症(endometriosis)之成因不明，一般相信和細胞外間質(extracellular matrix, ECM)的生成與退化有關；細胞外間質金屬蛋白酶(matrix metalloproteinases, MMPs)及細胞外間質金屬蛋白酶抑制劑(tissue inhibitors of metalloproteinases, TIMPs)於調節細胞外質的生成與退化上扮演重要角色。臨床上無快速診斷子宮內膜異位症的方法，治療上常用親生殖腺素釋放激素(gonadotropin-releasing hormone analog, GnRHa)或經腹腔鏡以電燒去除不正常附著的內膜組織。本研究希望自血液中找到診斷子宮內膜異位症之分子標記並瞭解子宮內膜異位症與MMPs及TIMPs的關聯及GnRHa對此類酵素的影響。利用西方墨點法發現71.9％未經GnRHa治療血漿檢體中偵測到游離型細胞外間質金屬蛋白酶抑制劑第一型(tissue inhibitors of metalloproteinases-1, TIMP-1)；只有27.6％經GnRHa治療血漿偵測到游離型TIMP-1。67%未經GnRHa治療腹腔液(peritoneal fluid, PF)亦偵測到游離型TIMP-1的存在。反轉錄聚合酶錬鎖反應顯示組織中細胞外間質金屬蛋白酶第九型(matrix metalloproteinases-9, MMP-9) 和TIMP-1 mRNA表現量無差異。子宮內膜異位症患者組織中細胞外間質表現經RT-PCR顯示未經GnRHa治療的異位內膜組織中膠原蛋白第一型(collagen type I)及小分子蛋白醣(proteoglycan)，decorin mRNA表現量比經GnRHa治療之組織多。免疫組織染色法(immunocytochemistry)顯示decorine及Biglycan在基質及上皮組織、內皮組織和血管腔壁的分布表現。本研究証實子宮內膜異位症病程與MMPs及TIMPs的表現有關，此類酵素及ECM表現受到GnRHa影響。血漿中游離型TIMP-1可作為早期診斷子宮內膜異位症之指標及預後之追蹤標記。

2. Differential Expression of Extracellular Matrix in Women with Endometriosis by Gonadotropin-Releasing Hormone • The pathology of endometriosis is not clear. It is believed that the extracellular matrix(ECM) remodeling is relevant to the progression of endometriosis. The combined action of matrix metalloproteinases(MMPs) and the tissue inhibitors of MMPs(TIMPs) plays important roles in ECM remodeling. Thus far, there is no clinical markers for early diagnosis of endometriosis, unless using the invasive operation laparoscopy to confirm. Gonadotropin-releasing hormone analog (GnRHa) or the laparoscopy to remove the adhered endometrium is the common treatment for endometriosis. However, the side effect such as osteoporosis and the discomfort by operation may occur. This study aims to search the potential serum markers for diagnosis of endometriosis and to understand the effect of GnRHa on MMPs, TIMPs and ECM remodeling in endometriosis. Western-blot analysis indicated that the sera in 23 out of 32 women(71.9%) with endometriosis not receiving GnRHa treatment have been detected the presence of free form tissue inhibitors of metalloproteinases-1(TIMP-1), whereas only 27.6% (8/29) sera with GnRHa treatment have been detected. The total concentration of TIMP-1 in serum measured by ELISA, however, was not significantly different between the patients with or without GnRHa treatment. Additionally, The activity of MMP9 was measured by ELISA as well. The results indicated that the activity of MMP9 in serum was significantly lower in the patients without GnRHa treatment. RT-PCR results showed that no significantly differences at the mRNA level for TIMP-1 and MMP9 in tissue between the patients. It suggested that the presence of TIMP-1 and decreased activity of MMP9 in sera in the endometriosis patients without GnRHa treatment may respond to the aberrant adherence of endometrium. To study the ECM composition in eutopic/ectopic endometrium, the results indicated that by RT-PCR analysis, the mRNA expression of collagen type I and the small proteoglycan, decorin, was more in the patients without GnRHa treatment compared to that in patients with GnRHa treatment. Immunocytochemistry study showed that decorin exist widely in stromal cells, epithelial cells and endothelial cells, whereas the distribution of biglycan is more restricted. To follow the matrix deposition in the eutopic endometrium during menstrual cycle, decorin and biglycan were expressed intensively in the stroma in the proliferation phase before ovulation and expressed in the epithelium in the secretory phase, suggesting that the hormone in deed regulated the matrix remodeling. In summary, the MMPs, TIMPs, and the expression of ECMs are affected by GnRHa. The presence of free form TIMP-1 in sera can been a useful marker for early diagnosis of endometriosis and the trace marker for prognosis.