Regulation The Expression Of Plasminogen Activator And Interluekin-6 In Women Plasma By Vitamin E After Abdominal Gynecological Operation
M Pawelec, Y Saleh, M Siewinski, A Karmowski, T Sebzda
interleukin-6, plasminogen activator upa, plasminogen inhibitor pai-1, vitamin e
M Pawelec, Y Saleh, M Siewinski, A Karmowski, T Sebzda. Regulation The Expression Of Plasminogen Activator And Interluekin-6 In Women Plasma By Vitamin E After Abdominal Gynecological Operation. The Internet Journal of Gynecology and Obstetrics. 2004 Volume 4 Number 2.
During the whole period of woman life there are many processes which involve degradation of extracellular matrix (tissue morphogenesis, angiogenesis, bone remodelling, wound healing, trophoblast implantation, involution of postpartum uterus or involution of postlactation mammary gland). Also, proteolysis is found in pathological conditions such as tumor growth, metastasis, arthritis and autoimmune diseases (1). Urokinase plasminogen activator (uPA) and urokinase plasminogen activator receptor (uPAR) are the central molecules for uPA/uPAR/plasmin-dependent proteolysis, which is thought to play a significant role in the development of pregnancy, as well as its many complications during pregnancy. The results support the hypothesis that the fibrinolytic system participates in preterm rupture of membranes and placental abruption (2). Aflalo
High intake of n-6 fatty acids may attenuate beneficial effects of n-3 fatty acids. These results suggest that the inhibition of inflammatory cytokines may be one possible mechanism for the observed beneficial effects of fatty acids on chronic inflammatory diseases (5). Son
Materials and Methods
Venous blood samples from the study (34 women undergoing abdominal gynecological operation: amputatio uteri, exstirpatio uteri or ovariectomia) and the control group (20 healthy women in the same age) were collected from the antecubical vein. The control group consisted of 20 healthy individuals who were invited for routine gynecological examination and Pap smear (complimentary). The study group was further divided into 2 subgroups: 17 women undergoing gynecologial operation for non oncological diseases (mainly myomata uteri - 16, and benign ovarian tumors - 1) and not treated with vitamin E after the operation and another 17 women undergoing gynecologial operation for non oncological diseases (mainly myomata uteri - 15 and benign ovarian tumors - 2) treated orally with vitamin E after the operation. Mean age in all groups was 48 years. All cases with processes involving fibrinolytic activation (described in literature) were excluded. Blood was collected in plastic tubes with 3.8 % sodium citrate about 9 a.m. The collected blood was mixed immediately by inverting the tube, cooled with ice-cold water and centrifuged at 2000 xg for 10 minutes to remove the plasma. The plasma samples were stored frozen (–80 °C) until used. 17 operated patients were treated orally with 800 mg vitamin E daily for 7 days (vitamin E was obtained from Hasco, Wroclaw, Poland). Another 17 operated patients were not treated with vitamin E. The blood from operated women was collected for the first time the day before operation, for the second time on the first day after the operation and for the third time on the 7th day after the operation.
The concentrations of uPA, PAI-1 and interleukin-6 were measured by enzyme immunoassay kits. The required dilutions, antibodies, conditions, and detection ranges for each ELISA were done according to protocol procedures (11). Serum level of IL-6 was measured with commercially available quantitative “sandwich” enzyme-linked immunosorbent assay (Quantikine) kits obtained from R&D System using specific antibodies. The concentration of each protein was measured at 450 nm on a microplate reader (Dynex Technologies, Billinghurst, UK). Values for the PAS components (ng/ml) and interleukin-6 (pg/ml) were determined for each sample from a standard curve using Revelation Software (Dynex Technologies, Billinghurst, UK). Final plasma values were expressed as ng/ml or pg/mg protein for PAS components and interleukin-6, respectively.
The amount of vitamin E was determined by liquid chromatography, using a HPLC apparatus (“Philips”) and a Pye Unicam PU 4020 UV detector. The results were decoded with use of Peak Simple Chromatography Data System Program (11).
Values are expressed as mean ± SD. The significance of the mean differences between groups was assessed by the Student's two-tailed unpaired t test. The correlation coefficient was determined by linear regression analysis. Differences were considered significant at p<0.05.
In our study, paired non oncological cases before operation compaired with haelthy individuals were analysed for expression of interleukin-6, uPA, and PAI-1. uPA values were significantly higher in non oncological gynecological diseases (median 2.5 ng/ml, range 0.6–7.3) than in healthy individuals (median 0.3 ng/ml, range 0–3.6;
Figure 1: Changes in uPA, PAI-1, and interleukin-6 concentrations in women plasma with non-oenological cases in comparison with control (healthy).
The significance of the differences in median values of patients with non-oncologecal cases and Healthy individuals women (control) were calculated by Wilcoxon matched pairs signed-rank test. Differences were considered significant at p<0.05.
Interleukin-6 values were: median 2.5 ng/ml, range 1.2-4.6 (
Figure 2: Changes in uPA, interleukin-6 and vitamin E concentrations in women plasma with non-oencological cases
The significance of the differences in median values of patients after operation and without treatment with vitamin E and patients after operation and after treated with vitamin E were calculated by Wilcoxon matched pairs signed-rank test. The correlation coefficient was determined by linear regression analysis. Differences were considered significant at p<0.05.
The female reproductive system seems to be protected from infection, destruction and oncogenesis. The objective of this study was to determine if there is stimulation of production of uPA, PAI-1 and interleukin-6 in women with non oncological gynecological diseases before and after operation. We also wanted to know whether vitamin E could inhibit this process. The different levels of uPA, PAI-1 and interleukin-6 in non oncological gynecological diseases and in healthy individuals confirm that the reproduction system is a rich source of these markers, and the levels of these markers change not only in oncological and inflammatory diseases. These levels are also changed in cases of myomata uteri and benign ovarian tumors. uPA is involved in serum degradation and PAI-1 in fibrinolysis (2). The balance between the expression of the activators and inhibitors is important in vivo in determining whether matrix degradation and activation of other proteinases is likely to occur. In our study, both uPA and the inhibitor PAI-1 were greater in women with non oncological diseases than in healthy individuals. However, previous studies showed that this increase of PAI-1 does not significantly inhibit uPA activity in primary tumours; therefore, the balance will favour proteolysis (13), which can also take place in our cases. Our results show that uPA, PAI-1, and interleukin-6 levels were significantly higher in non oncological gynecological cases in comparison with healthy individuals (
The cascade of proteolytic reactions triggered by the plasminogen activators is started by the conversion of the inactive proenzyme plasminogen into the active enzyme plasmin, a protease of wide specificity, which can directly cleave fibrin/fibrinogen, blood coagulation factors V/Va and VIII/VIIIa, some growth factors, and extracellular matrix components, and can also catalyze the degradation of plasmin-resistant matrix proteins, such as natural collagens, due to activation of inactive zymogens of collagenases (matrix metalloproteinases); thus, it can play the dominant role in extracellular proteolysis throughout the body (18,19, 20). The dominant role of plasmin was confirmed only within the last 15 years, when plasminogen synthesis and plasmin activation were reported to occur everywhere in the body (21). During the development process of neoplasmatic disease, the activity of these enzymes increases, they activate other proteolytic enzymes like elastase, kolagenase which causes catalytic degradation of healthy tissues by so doing initiate the development of a disease and their autogenic inhibitors are not able to stop the developing changes (22, 23). After abdominal gyneocological operation the patients treated daily with 800 mg vitamin E for 7 days had uPA and Interleukin-6 levels decreased in comparison with the group of patients without such treatment (
Yousif Saleh, PhD, MD 1st Clinic of Obstetrics and Gynaecology Medical University of Wroclaw Chalubinskiego 3 50-368 Wroclaw, Poland tel: +4871 7842413, e-mail: email@example.com