| Endometriosis is defined as the presence of endometrial glands and stroma outside the uterus and myometrium. It is a common benign, yet highly enigmatic gynecological disease affecting approximately10-15%of women of reproductive age. Symptomatic endometriosis causes long-term physical and mental sufferings and results in significant reduction in working ability and quality of life.Surgery or a combined surgical-medical technique is the treatment of choice for the management of endometriosis since the medical treatment alone is either inadequate or of side effects. Surgical removal of lesions has a better short-term effect. Despite the proven therapeutic efficacy of surgery, high recurrence after surgery provides a formidable challenge for gynecologists:the cumulative recurrence rate of the disease has been reported to be40-50%5years after the primary surgery. High recurrence rate and short time of recurrence has seriously affected the quality of life of patients. There, reduction of recurrence has of great clinical significance.Laparoscopic surgery has the advantages of less operative incision, less pain. shorter hospitalization, faster recovery and cosmetic results. Especially, it affords the benefits of good exposure and magnification. Based on the sufficient visual field and magnified visualization of endometriotic lesions provided by laparoscopy, conservative laparoscopic surgery has an outstanding advantage of completeness in removing all forms of endometriotic lesions which is difficult obtained during laparotomy. Currently, laparoscopic excision of endometriosis is considered to be the first-line method for the treatment. Despite the proven therapeutic efficacy of conservative laparoscopic surgery, recurrence of endometriosis is still high. It is reported that there is no difference about5-year cumulative recurrence rate between laparotomy and laparoscopy.As we all know, laparoscopy need continuous infusion of carbon dioxide to keep artificial pneumoperitoneum. Although CO2gas has a negative impact on cardiopulmonary function, it is considered to be the ideal gas to create a pneumoperitoneum during laparoscopic surgery because of the advantage of being cheap, colorless, noninflammable, absorbent, and excretive. CO2is soluble in water, reducing air embolism which is the most serious complication. CO2has active physiological function and has pathophysiological effect on body especially intra-abdominal environment. This forces me to think that if laparoscopy itself or CO2pneumoperitoneum has an effect on residual endometritic lesions after laparoscopic surgery and take part in the recurrence of endometriosis. As far as we know, no previous study on the behavior of residual endometriotic lesions exposed to CO2pneumoperitoneum has been reported.In recent years, it is reported that CO2gas is closely related to tumor environment, local immune, growth, implantation and metastasis. CO2gas may promote intra-abdominal tumor growth such as ovarian cancer, colon cancer and liver cancer through acidosis, low temperature, drying and mechanical pressure. It is positively correlated to pressure and action time. CO2gas can induce expression of cognate molecules with metastasis, angiogenesis factors, matrix metalloproteinases and inhibit the number of abdominal immune cells and activity. Studies have found that CO2gas itself can stimulate the growth of tumor cells.As we all know, endometriosis is considered to be a benign disorder but it contains some features similar to malignant tumors such as invasion, neoangiogenesis, recurrence and metastases. The balance between proliferation and apoptosis was broken when endometriosis occurred.Based on our hypothesis, we design this study. First of all, we successfully establish primary cell culture method of endometriotic cells, make CO2pneumoperitoneum experimental device, and then mimic CO2pneumoperitoneum with different pressure and action time. Therefore, our study was designed to in vitro investigate the growth and apoptosis of primary cultured endometriotic cells after exposure to a100%CO2pneumoperitoneum environment simulated laparoscopy at different pressures. We choose the strongest influence pressure point and processing time. Finally, we analyze associated protein through protein array. These findings may contribute to reducing or avoiding recurrence of endometriosis after laparoscopic treatment. Part â… Influence of carbon dioxide pneumoperitoneum mimicking laparoscopic surgery on proliferation and apoptosis of human endometriotic cellsObjectives:To investigate the effects of a simulated carbon dioxide (CO?) pneumoperitoneum environment during laparoscopy on proliferation and apoptosis of primary cultured human endometriotic cells at different pressures.Methods:Endometriotic cells were isolated and cultured. Passaged endometriotic cells were exposed to a100%carbon dioxide environment at different pressures (5mmHg,10mmHg,15mmHg) for different times (1h,2h) respectively. After dealed with carbon dioxide pneumoperitoneum, cells were observed under inverted microscope and transmission electron microscopy. Cell cycle was determined by flow cytometry. Apoptosis was measured by FITC labeled Annexin-V and PI double staining and Terminal deoxynucleotidyl transferase-mediated dUTP-fluorescein end labeling (TUNEL) assays.Results:1. Primary cultured endometriotic stromal cells are spindle and gland cells are spiral, gland cells are rare. After passaged,99%cells are stromal cells.2. pH of culture medium in carbon dioxide pneumoperitoneum group decreased significantly compared with control group especially in high pressure group (p<0.05)3. There were no significant difference in proliferative index between carbon dioxide pneumoperitoneum and control group (p>0.05)4. Apoptitic index was lower after15mmHg pressure carbon dioxide pneumoperitoneum for2h than control group (p<0.05)Conclusions:1. Endometriotic cells cultured by collagenase and DNase I digestion method provide a more ideal cell model for study of endometriosis.2. Carbon dioxide (CO2) pneumoperitoneum device in vitro simulate the process of laparoscic surgery successfully.3. Carbon dioxide (CO2) pneumoperitoneum may take part in the recurrence of endometriosis through inhibiting endometriotic cells apoptosis at a higher pressure. Part â…¡Altered protein expression profiles of endometriotic cells dealed with carbon dioxide pneumoperitoneumObjectives:To investigate the changes of apoptosis related protein expression profile between control group and carbon dioxide pneumoperitoneum group in order to detect differential proteins.Methods:Endometriotic cells were isolated and cultured. Passaged endometriotic cells were exposed to a100%carbon dioxide environment at15mmHg for2h. After48h cultured in normal condition, cells were tested by RayBio(?) Human Apoptosis Antibody Array I to choose apoptosis-related protein. Differential proteins were further validated by using real-time PCR and western blot analysis.Results:Eight proteins were up-or down-regulated in carbon dioxide pneumoperitoneum group. XIAP was validated by western blot analysis.Conclusions:1. There were differential apoptosis related proteins between control group and carbon dioxide pneumoperitoneum group.2. Carbon dioxide pneumoperitoneum may take part in endometriosis recurrence through inhibitors of apoptosis protein such as XIAP.
|
PDF Full Text Request