Serum Before And After Laparoscopic And Open Surgery Of Ecv304 Generate Endothelial Nitric Oxide Synthase

Subject: the impact of laparoscopic surgery and laparotomy perioperatively in serum on the level of endothelial nitric oxide synthase generated by ECV304Objective: By observing the changes of eNOS level in serum of patients who had received gynecological laparoscopic surgery or laparotomy, we could explore whether the specific factor in gynecological laparoscopic surgery would contribute to the damage of vascular endothelial cells, and accordingly increase the danger of phlebothrombosis. Simultaneously, culturing the human umbilical vein endothelia cell line (ECV304) in patients'serum to observe the alteration of cell appearance and the synthesis level of eNOS.Methods: To decide serum as example, through enzyme linked immunosorbent assay,we detected the content of eNOS in 8 patients'serum and blood plasma which were both not inactivated, confirmed serum that not inactivated to be the experimental specimens source.(1). Select 19 patients who have received laparoscopic hysterectomy as study group, accordingly, select 19 patients that have received abdominal hysterectomy as matched group, detect the content of eNOS of patients'venous respectively at preoperative and the end of surgery to observe the impact of laparoscopic surgery and laparotomy perioperatively on the level of eNOS.(2). Choose serum from 16 cases (laparoscopic hysterectomy, 8 cases, abdominal hysterectomy 8 cases) before operation and after operation as interfere media to culture ECV304 in vitro, and observe the alteration of cell shape as well as the content variation of eNOS through ELISA. The results measurement data using T-test, single-factor analysis of variance, the sample mean variance analysis (F test) and using SPSS17.0 to analyse statistically.Results:1. Endothelial nitric oxide synthase concentration of not inactivated serum/ inactivated serum from one patient is not statistically significant. (P>0.05)2.(1) Compared with preoperative, the concentration of eNOS decreased slightly but not statistically significant in both groups postoperatively. (P>0.05) (2)The difference of eNOS during operation was not statistically significant between groups. (P>0.05)3.(1)The control group which is ECV304 cell line without intervention serum to culture for 2h,6h,12h,24h, then it is no significant when detecting the level of eNOS in cell culture medium. (P>0.05)(2)Serum gained from patients before surgery and after in both study group and control group to control in cell culture medium 2h,6h,12h,24h, the level of eNOS in supernatant decreased slightly than the basic level of eNOS in serum which is not intervention. However, there is no significant. (P>0.05). comparing the difference amount of eNOS in each group at different times, it is no significant. (P>0.05).(3) Serum gained from patients before surgery and after in study group which is ECV304 cell line to culture for 2h,6h,12h,24h, comparing the level of eNOS in intervention serum, it is no significant. (P>0.05).(4) Serum gained from patients before surgery and after in control group which is ECV304 cell line to culture for 2h,6h,12h,24h, comparing the level of eNOS in intervention serum, it is no significant. (P>0.05).(5) After subcultured of ECV304 cell, cells were mostly spindle and irregular triangle with the characteristic of cobblestone-like and growth single, cell growth adherent state clearly. Nuclei were round or oval with rich cytoplasm. There is a small protuberance protruding from the cell formed filaments with membrane integrity and cytoplasm transparent. There is phenomenon of cell accumulation. The ECV304 cells possess the characteristic of endothelial cells with some features of the general. Affecting 24h by intervention serum , observed the cell morphology in study group and control group using inverted phase contrast microscope that the cells became round and off at different degrees .Conclusion:(1) Not inactivated serum can be used as specimens.(2) Compared with open surgery,laparoscopic surgery did not lead to decrease Endothelial nitric oxide synthase.(3) The concentrations of Endothelial nitric oxide synthase by cultured endothelial cells did not decrease after the activation of serum from laparoscopic surgery patients.(4) Laparoscopic surgery on patients with no thrombosis risk factors did not increase the possibility of lower extremity venous thrombosis.