| Endometriosis is a common disorder on gynecology, that affects up to 15% of all women of reproductive age, although not neoplastic, it shares the features with malignant cells and the mechanisms proposed for micrometastasis in cancer. The clinic finding of endometriosis is characterised by pelvic pain and infertility. Our current knowledge about the pathogenesis, pathophysiology of the pain and related infertility is still limited. At present, laparoscopy is still considered the gold standard in diagnosis of endometriosis. But there still has been no a perfect approach for management of the disorder. Complete resolution of endometriosis is not yet possible but therapy has essentially three main objectives: 1) to reduce pain; 2) to increase the possibility of pregnancy; 3) to delay recurrence for as long as possible. Surgical treatment is considered the best treatment for women with pain and or pelvic mass who wish to become pregnant in a short time soon. Although the absence of circulating oestrogen levels leads to a regression of the disease, it also induces many unpleasant side effects and reduces life quality. In recent years numerous investigations have implicated that disturbances in theimmune response is fundamental to its etiology and pathogenesis. Immune alterations including increased number and activation of peritoneal macrophages, decreased T cell reactivity and natural killer cell cytotoxicity, increased circulating antibodies, and changes in the cytokine network indicate that immunologic defect or imbalance plays an important role in the pathogenesis of endometriosis and endometriosis-associated infertility. Accumulating researches demonstrate that tumor necrosis factor alpha (TNF-alpha), as a cytotoxic product of activated monocytes, macrophages, and lymphocytes, involves in the development and maintenance of endometriosis and associated infertility. TNF-alpha may be thought as a useful marker for both activity and stage of endometriosis. At the meantime, TNF-alpha may be a marker for both activity and stage of endometriosis. So TNF inhibitors may be an attractive alternative to the therapeutic regimens, their benefits may indicate a future potential therapeutic targets for endometriosis.In this study, we intend to realize the effect of TNF-alpha on endometrial cells from women with endometriosis and further to understand the mechanism of endometriosis from molecular, cellular, and pathophysiological aspects. At the sametime, we tried to find new target and pathway for biological therapy of endometriosis. At first step, we established cell culture models of endometrium (including normal from control, eutopic, and ectopic endometrium from patients withendometriosis) and identified them at the molecular and cellular level; then we studied the effect TNF-alpha in dosage course and soluble tumor necrosis factor-alpha receptor type l(sTNFR I ) on endometrial stroma cell.PART I Study of glandular epithelial cell and stromal cell cultureof human endometrium in vitroObjctive To establish a method of isolation, purification and culturing of human glandular epithelial and stromal cells of normal, eutopic, and ectopic endometrium in vitro. (NE ,EU,EC )Methods Samples of endometrium were obtained from patients undergoing gynecological surgery and lapalrotomy for benign condition and endometriosis. Endometrium was minced and digested with 0.125% collagenaseⅣ. Single stromal cells and epithelial glands were separated by filtration through 250μm and 38μm (or 74μm) mesh, stromal cells and epithelial glands were cultured in complete culture medium. After 30 ~ 60 minites, Culture medium of stromal cells was changed and unfixed stromal cells and red blood cells were wiped off. The culture medium was changed every 2 ~ 3 days. Stromal cells and epithelial glands were identified by immunohistochemistory with vimentin and cytokeratin. The cells were digested by 0.25% trypsin and0.02%EDTA after nearly 90% of them were merged. Result The single stromal cells attach to the wall in 30 ~ 60 minites and grow into fi... |
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