| BachgroundEndometriosis (EMS) is histologically defined by the presence of endometrium-like glands and stroma outside the uterus. EM is often associated with pelvic pain, dysmenorrhoea and infertility, in around 10%-15% of women of reproductive age. Despite its high prevalence in the female population, the etiology and physiopathology of the disease are not fully elucidated and few effective therapeutic approaches are available for women suffering from the disease. Integrin a v β 3 is the glycoprotein of the cell surface, regulating biological events which including adhesion of cell to extracellular matrix, platelet aggregation, immunal function, tissue repair, tumor infiltration, angiogenesis and so on. The metastasis of tumour cells is related with active integrin a v β 3. It is known that the cell of endometrium of endometriosis is similar to the tumour cell at the biology behavior ,liking attachment, aggression and angiogenesis.It will be a new way to the treatment of endomeriosis by blocking this process. We study the effect of monoclonal antibody to integrin a v β 3 on the treatment of endometriosis in this paper. ObjectivesStudy the influence of monoclonal antibody to integrin a v β 3 (Lm609, 7e3) on the attachment of cells of endometrium of endometriosis. Study the inhibition of Lm609 on angiogenesis after ectopic implantation of human endometrium of endometriosis on the chicken chorioallantoic membrane. Establishing the endometriosis animal model of SCID mouse, study the effect of Lm609 on the formation of ectopic lesion.Materials and methods1, Obtain three human samples of eutopic endometrium of endometriosis. Culture the cells of eutopic endometrium and diluted to obtain a final concentration of 1×104/ml. The cells add to fibronectin-coated 96-well microtiter plates 100ul every well. Treated groups add diluted antibodies to integrin a v β3 (Lm609, 7E3) to the wells in concentration of 80ug/ml, 40ug/ml, 20ug/ml, 10ug/ml and control group add PBS at the same volume. Every concentration repeated three wells. After incubated 1 hour at 37℃, add 15ul MTT every well. After 4 hour at 37℃, the cell suspensions were removed and replaced by 100ul DMSO every well. Measure the optical density (OB) at the wavelengths of 492 nm. Every sample repeated three times. 2, Obtain one human sample of eutopic endometrium of endometriosis and gently trimmed into small pieces(1mm3). Purchased 60 nigh-day-old chick embryos. A small hole was made on the broad side of the egg directly over embryonic blood vessels. Divided the eggs into three groups, one group of chicken chorioallantoic membranes received sample pieces and 50ug Lm609, the other group received sample pieces and 100ul PBS and the last group received 100ul PBS. Take the chicken chorioallantoic membranes two days later.3 , Obtain four human samples of eutopic endometrium of endometriosis and gently trimmed into small pieces(1mm3). Mix these pieces with PBS into tissue suspensions. The eutopic endometrium was injected into the subcutaneous of abdomen of SCID mice. The mice divided into two groups one week later, one group treated with PBS and the other group treated with Lm609 two times each week. Kill all mice and take the samples of lesions after three weeks. Results 1 , OB of the control group is higher than the treated groups(P<0.05). There is nodifferent between every concentration(P>0.05). 2, The CAM angiogenesis of the control group is stronger than empty control... |
