Activation of PAR-1/NADPH oxidase/ROS signaling pathways is crucial for the thrombin-induced sFlt-1 production in extravillous trophoblasts: possible involvement in the pathogenesis of preeclampsia.

BACKGROUNDS/AIMS: Preeclampsia was characterized by excessive thrombin generation in placentas and previous researches showed that thrombin could enhance soluble Fms-like tyrosine kinase 1 (sFlt-1) expression in first trimester trophoblasts. However, the detailed mechanism for the sFlt-1 over-production induced by thrombin was largely unknown. The purpose of this study was to explore the possible signaling pathway of thrombin-induced sFlt-1 production in extravillous trophoblasts (EVT). METHODS: An EVT cell line (HRT-8/SVneo) was treated with various concentrations of thrombin. The mRNA expression and protein secretion of sFlt-1 in EVT were detected with real-time polymerase chain reaction and ELISA, respectively. The levels of intracellular reactive oxygen species (ROS) production were determined by DCFH-DA. RESULTS: Exposure of EVT to thrombin induced increased intracellular ROS generation and overexpression of sFlt-1 at both mRNA and protein levels in a dose dependent manner. Short interfering RNA (siRNA) directed against PAR-1 or apocynin (an inhibitor of NADPH oxidase) could decrease the intracellular ROS generation and subsequently suppressed the production of sFlt-1 at mRNA and protein levels. CONCLUSIONS: Our results suggested that thrombin increased sFlt-1 production in EVT via the PAR-1 /NADPH oxidase /ROS signaling pathway. This also highlights the PAR-1 / NADPH oxidase / ROS pathway might be a potential therapeutic target for the prevention of preeclampsia in the future.

[Relationship between pigment epithelium-derived factor expressed in placentas and the pathogenesis of preeclampsia disease].

OBJECTIVE: To investigate the effect of pigment epithelium-derived factor (PEDF) on the pathogenesis of preeclampsia disease, by detecting the expression of PEDF in the placentas, as well as the relationship between PEDF and the production of placental vessels. METHODS: A study was performed in 60 cases of pregnant women with preeclampsia in the obstetrical department of Nanfang Hospital affiliated to southern medical university from October 2011 to January 2013, in which 30 cases were patients with mild preeclampsia(mPE) and other 30 cases were those with severe preeclampsia (sPE).40 normal pregnant women who also been hospitalized and delivered were selected as control group. The expression of PEDF and micro-vessel density (MVD) in placentas were assayed by using western blot and SP immunohistochemical method, then the relationship between PEDF and MVD was analyzed. RESULTS: (1) The pathological changes of placentas:the placental weight were lightened obviously in the mild preeclampsia and severe preeclampsia groups, the reduced blood vessels and luminal stegnosis were found in chorionic villus, basement membrane of trophocytes were thickening. The hyperplasia syneytiotrophoblast were like nodosity, with focus infarction, fibrinoid necrosis, or thrombogenesis.While there was no the above mentioned pathological alteration in normal control group. (2)The levels of PEDF expression in mild and severe preeclampsia group were 0.63 ± 0.09, 0.93 ± 0.07, while 0.47 ± 0.04 in control group, which in mild and sever preeclampsia groups were significantly higher than that in normal group (P < 0.05). Compared to mild preeclampsia group, the expression of PEDF was significantly increased in severe preeclampsia group, there was statistical significance between the difference (P < 0.05).(3) The amount of microvessel density (MVD) in mild and severe preeclampsia group were 106 ± 9, 93 ± 8, while 136 ± 9 in control group, which were significantly reduced in mild and severe preeclampsia group, compared to that in normal control group (P < 0.05). And it was significantly lower in severe preeclampsia group than that in mild preeclampsia group (P < 0.05). (4) The expression of PEDF was negatively correlated with the amount of MVD in mild and severe preeclampsia group (r = -0.426, P < 0.05; and r = -0.646, P < 0.05 respectively), which was also negative in control group (r = -0.589, P < 0.05). CONCLUSION: Increased PEDF expression in placentas of women with preeclampsia induce the dysfunction of the placental vascular reconstruction and the pathological alteration like ischemic and hypoxia in placentas, which may be involved in pathogenesis and pathogenic progress of preeclampsia.

[Diagnostic value of CA125, HE4 and Copenhagen Index in differentiating benign from malignant epithelial ovarian tumors].

OBJECTIVE: To analyze diagnostic value of Copenhagen Index based on pretreatment serum CA125, HE4 and age in differentiating benign and malignant epithelial ovarian tumors. METHODS: The clinical data were analyzed for 208 consecutive patients with epithelial ovarian tumors (including 100 with malignant and 108 with benign tumors) treated in our department between September, 2014 and September, 2016. The receiver-operating characteristic curve was drawn based on the golden standard of pathological diagnosis for calculation of the diagnostic sensitivity and specificity of CA125, HE4 and the Copenhagen Index. RESULTS: In the overall cases, early stage cases and advanced stage cases, the prediction probabilities of CA125, HE4 and Copenhagen Index were all significantly higher for malignant than in benign tumors (P<0.001). The sensitivities of CA125, HE4, Copenhagen Index for differentiating benign and malignant tumors were 81.0%, 86.0% and 91.0% in the overall cases, 64.0%, 68.0% and 72.0% in early stage cases, and 86.7%, 92.0% and 97.3% in advanced stage cases, and their diagnostic specificities were 88.0%, 93.5% and 96.3%, respectively. Copenhagen Index had the highest diagnostic sensitivity (but not in early stage cases) and specificity followed by HE4 and then by CA125 (P<0.001) (P>0.05). CONCLUSION: Copenhagen Index combined with CA125, HE4 and age hase better diagnostic value than HE4 or CA125 alone for differentiation between benign and malignant epithelial ovarian tumors, and can be used clinically to improve the early diagnostic rate of epithelial ovarian cancer.

[Effects of docosahexaenoic acid on cell apoptosis, invasion and migration of cervical cancer cells in vitro].

OBJECTIVE: To investigate the effect of docosahexaenoic acid (DHA) on apoptosis, migration and invasion of cervical cancer cell lines. METHODS: cervical cancer cell lines Hela and Siha in logarithmic phase were treated different concentrations of DHA. The morphological changes of the cells were observed microscopically and cell apoptosis was observed using Hoechst 33258 fluorescent staining. MTT assay was used to evaluate the effect of DHA in suppressing cell growth, and flow cytometry was employed to analyze the changes of cell apoptotic rate following DHA stimulations. Wound healing assay and Transwell migration assay were used to evaluate the migration of the cell lines. The expression levels of Bax, Bcl-2 cleaved caspase3, MMP-9 and VEGF proteins were detected by Western blotting. RESULTS: DHA exposure of the cells caused obvious morphological changes and dose-dependently increased the number of apoptotic bodies in the cells. MTT assay showed that DHA inhibited the growth of the cancer cells in a time- and concentration-dependent manner. DHA also effectively suppressed migration and invasion of the cancer cells. The cells exposed to DHA showed significantly down-regulation of Bcl-2, MMP-9 and VEGF proteins and up-regulation of cleaved-caspase 3 and Bax. CONCLUSION: DHA can promote cervical carcinoma cell apoptosis by down-regulating the anti-apoptotic proteins Bax, Bcl-2 and cleaved-caspase3 and suppress cell invasion by decreasing MMP-9 and VEGF expressions.