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Neurokinin B/neurokinin B Receptor-3 Act Through Activating p38MAPK Signal Pathway to Pre-eclampsia

Author: LiuYanYan
Tutor: ChenHanPing
School: Huazhong University of Science and Technology
Course: Obstetrics and Gynaecology
Keywords: Preeclampsia Placenta Plasma Neurokinin B Neurokinin B receptor p - p38MAPK Early pregnancy trophoblast cell line Hypoxia NKB/NKR3 p38MAPK SB203580 Flow cytometry MTT TEV - 1 NKB Invasion index
CLC: R714.245
Type: PhD thesis
Year: 2010
Downloads: 121
Quote: 0
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Purpose of a study in normal pregnancy, preeclampsia placental tissue neurokinin B (neurokinin B NKB), and its receptor (neurokinin B receptor, NKR) in the expression of the protein and mRNA levels, and normal pregnancy, preeclampsia pregnant women plasma neurokinin B peptide content, to take this on neurokinin B and pre-eclampsia relationship; p38MAPK activation protein in normal and preeclampsia, and to explore of NKB with the p-p38MAPK expression related sex. Methods normal pregnancy, mild preeclampsia, severe preeclampsia and 20 cases by enzyme-linked immunosorbent assay (ELISA) quantitative detection in maternal plasma NKB content; Yihong - hematoxylin (HE) staining, light microscopy normal pregnancy and preeclampsia pathological changes of the structure; NKB in / NKR, p38MAPK protein localization and quantitative expression detected by immunohistochemistry in placental tissue; reverse transcription polymerase chain reaction (reverse transcription polymerasechain reaction, RT-PCR) detection NKB in placental tissue / NKR mRNA expression. Results mild and severe preeclampsia in maternal plasma level of NKB was significantly higher than in normal pregnant women; 2. Normal placental NKB / NKR small amount of the distribution in the syncytiotrophoblast, capillary endothelial cells, mainly expressed in the cytoplasm and membrane , a small amount of p-p38MAPK located in trophoblast cells, capillary endothelial cells, mainly expressed in the nucleus; normal pregnancy placenta, mild preeclampsia, severe preeclampsia group NKB expression were: 0.078 ± 0.005,0.082 ± 0.011,0.101 ± 0.022, light, severe preeclampsia placental NKB protein expression compared with normal pregnancy was significantly higher (P lt; 0.05, P lt; 0.01); normal pregnancy, the placenta, mild preeclampsia, severe preeclampsia group NKR expression is: 0.069 ± 0.054,0.076 ± 0.110,0.090 ± 0.120, light, moderate and severe preeclampsia NKR protein expression compared with normal pregnancy was significantly higher (P lt; 0.05, P lt; 0.05); normal pregnancy, the placenta, mild sub- preeclampsia, severe preeclampsia group p-p38 MAPK protein were: 0.052 ± 0.012,0.069 ± 0.054,0.094 ± 0.047, light, p-p38 MAPK protein expression in severe preeclampsia compared with normal pregnancy was significantly higher (P lt; 0.05, P lt; 0.01),; and normal pregnancy placental NKB and p-p38MAPK expression was positive related (r 0.503, p lt; 0.05); mild and severe preeclampsia group, NKB in with p-p38MAPK level was also found positive related (r = 0.515, p lt; 0.05 and r = 0.657, p lt; 0.01); normal placental NKB with NKR3 mRNA expression levels are: 0.560 ± 0.248,0.441 ± 0.206, preeclampsia NKB and NKR3 expression levels were significantly increased, NKB in mild preeclampsia group to 0.772 ± 0.142, 0.954 ± 0.237 NKR3 mild preeclampsia severe preeclampsia group to 0.578 ± 0.253, severe preeclampsia group was 0.804 ± 0.316 . Conclusion NKB / NKR expression increased pre-eclampsia is closely related to p-p38MAPK and NKB / NKR regulated expression of mutual influence, may be one of the mechanisms involved in the pathogenesis of preeclampsia. Purpose 1. Study hypoxia in human early pregnancy trophoblast cell line (TEV-1) NKB/MKR3 expression; and hypoxia, the kinase activity of p38 MAPK inhibitor SB203580 on P38MAPK, from the cellular level to explore NKB/MKR3, of P38MAPK with the relationship between pre-eclampsia; research hypoxia of SB203580 intervention on TEV-1 cell line apoptosis and proliferation of change-depth study of hypoxia, the role of p38 MAPK on the regulation of trophoblastic biological characteristics. Method 1. TEV-1 cells chemical hypoxia induced by CoCl2 in vitro simulation the trophoblastic vitro hypoxia model, mRNA expression using RT-PCR detection of TEV-1 in NKB/NKR3; were with COCl2 and SB203580 intervention TEV-1 cell lines to study the TEV-1 cells P38MAPK kinase activity changes; cobalt dichloride, SB203580 in vitro intervention TEV-1 cell lines, flow cytometry apoptosis rate of the different time points, MTT (Methl thiazolyl tetrazolium, MTT) colorimetric determination of trophoblastic proliferation and at different time points. 1 With the increase of hypoxia, of trophoblastic NKB/NKR3 mRNA expression gradually increased to 24 hours, reached the peak hypoxia 1h, 6h, 12h significantly increased compared with normal cultured 24h compared to significantly liter high, followed by a slow decline; CoCl2 treatment trophoblastic 20 minutes after a dose-dependent activation of p38MAPK and p38MAPK selective inhibitor SB203580 in a concentration-dependent inhibition of p38MAPK kinase activity; With hypoxia extend nourish early apoptosis rate was gradually increased, especially from 24h (24h, 48h, 72h) compared with the normal culture, apoptosis rate was significantly higher, while a p38MAPK the selective inhibitor SB can be reduced hypoxia-inducible trophoblastic early apoptosis rate increase; extend the with the anoxic intervention time, the ability of Sertoli cells to proliferate are subject to varying degrees of inhibition, cultured for 24 h inhibited trophoblast cell proliferation is the most obvious, and p-p38 MAPK inhibitor SB weakened CoCl2 inhibited the proliferation of trophoblastic effect. Conclusion 1. Hypoxia can induce TVE-1 cells high expression NKB/NKR3, also showed a concentration-dependent activation of p38MAPK kinase activity; 2. Increase with hypoxia time trophoblastic apoptosis rate gradually increased proliferation gradually decreased, while the SB can be reduced both the role of hypoxia on trophoblast cells, the p38MAPK pathway play an important role in hypoxia-induced increase in trophoblast cell apoptosis rate and proliferation decreased. Objective To study the NKB in SB203580 on TEV-1 cell line invasion force, and explore NKB in possible regulatory mechanism of the role of p38 MAPK in pre-eclampsia. Method 1. Research NKB and SB203580 on p38MAPK kinase activity; the study concentration NKB different intervention time TEV-1 cell line invasiveness of; 3. Research SB203580 TEV-1 cell line invasiveness of. 1 trophoblasts NKB concentration-dependent activation of p38MAPK and a p38MAPK the selective inhibitor SB203580 in a concentration-dependent inhibition of NKB activation of p38MAPK; dose-dependent analysis showed that different concentrations of NKB for 48 hours, nourish cell invasion index decreased with the increased concentration of NKB 0.8mg / L NKB role in trophoblast cell invasion force after 48 hours, the weakest; analysis showed a time-dependent manner, by 0.8mg / L NKB respectively 12, 24, 48 hours, trophoblastic invasion index with NKB intervention of extension of time and reduce intervention for 48 hours, the weakest of trophoblast invasion force; 3.NKB inhibit TEV-1 cells in vitro invasion of; given alone SB203580 can inhibit trophoblast invasion, SB203580 also significantly inhibition weakened NKB trophoblast invasion. Conclusion trophoblasts NKB concentration-dependent activation of p38MAPK in a dose and time-dependent inhibition of TEV-1 cell invasion force, p38MAPK inhibitor SB203580 can be reduced NKB trophoblastic invasion inhibition. Description the NKB available through the p38MAPK pathway in trophoblast cells in vitro invasion.

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CLC: > Medicine, health > Obstetrics and Gynaecology > Obstetrics > Pathological pregnancy ( abnormal pregnancy ) > Gestosis > Eclampsia
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