Neurokinin B/neurokinin B Receptor-3 Act Through Activating P38MAPK Signal Pathway To Pre-eclampsia

Objective 1. To investigate the expression of neukinin B(NKB)/neuokinin B receptor(NKR) in placentas obtained from normal pregnancy and preeclampsia, and the relationship between NKB and preeclampsia; and the plasma NKB levels from normal and preeclampsia pregnant women;2. To investigate the the correlation of NKB and p-p38MAPK on the pathogenic mechanism of preeclampsia by detecting the expression of activating p38MAPK in placentas obtained from normal pregnancy and preeclampsia pregnant women;Methods 1.20 nomal pregnant women,20 mild preeclampsia, and 20 severe preeclampsia women were enrolled in the study, enzyme-linked immunosorbent assay (ELISA) was used to detect the expression of plasma NKB levels from normal and preeclampsia women;2. HE staining observed the pathology changes in placental tissues from normal and preeclampsia women; Immuno-histochemistry observed the expression of NKB/NKR, p-p38MAPK protein in placentas, and reverse transcript polymerase chain reaction (RT-PCR) were used to detect the expression of NKB/NKR3 mRNA in placental tissues from 20 normal pregnant women and 40 preeclampsia patients.Result 1. The plasma NKB level in the preeclampsia was significantly higher than that of normal pregnancy;2. Under the microscopic examination, nodules of syncytiotrophoblast, fibrinoid necrosis and damage of umbilical venous endomembrane were observed by HE staining. However, these microstructural changes are rarely found in the normal placenta; The positive staining of NKB/NKR were mainly located at cell membrane, and cytoplasma of trophoblast, p-p38MAPK protein were mainly located at nuclear membrane of trophoblast. The levels of NKB/NKR expression of placenta from preeclampsia patients were significant higher than that of normal group, the levels of p-p38MAPK protein expression of placenta from preeclampsia patients were higher than that of normal group; and there was obvious positive correlation between NKB and p-p38MAPK protein from normal and preeclampsia women;3. Contrast with normal group, the expression of NKB and NKR3 mRNA in preeclampsia group was significantly higher.Conclusion The increase levels of NKB/NKR may play an important role in the pathogenic mechanism of preeclampsia. The activation of p38MAPK signal pathway interaction with the over expression of NKB may play an important role in the pathgenic mechanism of preeclampsia.Objective 1. Investigating the effect of CoCl2 induced hypoxia on the expression of NKB/NKR3 of human first-trimester extravillous trophoblast cell line(TEV-1) in order to evaluate the role of NKB/NKR3 in preeclampsia. To investigate the effects of the activation of p38MAPK which was intervened by hypoxia, inhibition of p38MAPK(SB203580);2. To investigate the effects of CoCl2 induced hypoxia, SB203580 on the apoptosis and proliferation of trophoblast in order to further study the regulation of NKB and p38MAPK on the biological characteristics of trophoblast.Methods 1. TEV-1 was cultured respectively in normal and hypoxia circumstance which induced by CoCl2. RT-PCR was used to detect expression of NKB/NKR3 mRNA of TEV-1; the activation of p38MAPK was detected by cell ELISA after the intervention of CoCl2 and SB203580;2. TEV-1 was cultured respectively in CoCl2 and SB203580 circumstance, flowcytomery was performed to evaluate apoptosis of TEV-1 at different time, methl thiazolyl tetrazolium(MTT) was performed to evaluate proliferation of TEV-1 at different time.Result 1. The levels of NKB/NKR3 mRNA in TEV-1 of both groups increase gradually as the culturing time goes on, and they all go to the top at the 24-hour cultured time. Hypoxia could up-regulate the expression of NKB/NKR3. In TEV-1 cells, p38MAPK was activated by CoCl2 in a dose-dependent manner, and SB203580 could weaken the p38MAPK activation also in a dose-dependent manner;2. Under the hypoxia condition the trophoblast apoptotic rate was increase with time goes on. They all go to the top at the 24-hour cultured time, and after that decrease very tardily, and the inhibition of p38MAPK(SB203580) could weaken this effect; Under the hypoxia condition the trophoblast proliferation was decrease with time. They all go to the lowest point at the 24-hour cultured time, and after that decrease very tardily, and the inhibition of p38MAPK(SB203580) could also weaken this effect.Conclusion 1. CoCl2 simulated hypoxia up-regulate the expression of NKB/NKR3 and activated p38MAPK in a dose-dependent manner;2. The trophoblast apoptotic rate was increase and the proliferation was decrease with time goes on, and SB could weaken these effects. p38MAPK signal pathway may have an important role in the hypoxia induced the increase of trophoblast apoptotic rate and the decrease of trophoblast proliferation. Objective To investigate the effects of human recombinant NKB protein, the inhibition of p38MAPK(SB203580) on the invasiveness of TEV-1, in order to further study the regulation mechanism of NKB and p38MAPK on the biological characteristics of trophoblast.Methods 1. Cell ELISA was used to detected the activation of p38MAPK after the intervention of CoCl2 and SB203580;2. The TEV-1 cell's invasiveness was detected by transwell at different concentration and different time culture of NKB;3. The effect of SB on TEV-1 cell's invasiveness was detected by transwell.Result 1. NKB up-regulate the activation of p38MAPK in a dose-dependent manner, and SB203580 could weaken the p38MAPK activation also in a dose-dependent manner;2. The trophoblast invasiveness was decrease with the concentration of NKB increase, and the trophoblast invasiveness was decrease with the NKB cultured time goes on;3. NKB could inhibit the in vitro invasion of TEV-1; SB203580 solely could increase the in vitro invasion of TEV-1; but NKB and SB203580 combined will decrease the in vitro invasion of TEV-1 obviously.Conclusion NKB up-regulate the activation of p38MAPK in a dose-dependent manner, inhibit the in vitro invasion of TEV-1 in the dose-dependent and time-dependent manner; the inhibition of p38MAPK(SB203580) could weaken the inhibition invation of TEV-1. NKB coule regulate the trophoblast invasion in vitro by p38MAPK signal pathway.