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The Effects of Leptin on Invasiveness of Cytotrophoblasts and Injury of Endothelial Cell and the Relationship between These Effects and Pre-eclampia

Author: WuZuoZuo
Tutor: QiaoFuYuan
School: Huazhong University of Science and Technology
Course: Obstetrics and Gynaecology
Keywords: Preeclampsia Leptin TNF - alpha Adiponectin Early pregnancy trophoblast cell line Hypoxia MTT The invasion of Human umbilical vein endothelial cell line
CLC: R714.246
Type: PhD thesis
Year: 2009
Downloads: 187
Quote: 1
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Abstract


The purpose of the first part of the expression of leptin in the serum of normal pregnancy and preeclampsia and placental tissue. Study in normal pregnancy and preeclampsia serum and placental tissue leptin protein and mRNA expression differences, to explore the relationship between leptin and pre-eclampsia. 2. Detection of TNF-alpha and adiponectin serum in normal pregnancy and preeclampsia and placental tissue expression, and explore their correlation of leptin in the pathogenesis of preeclampsia. Method 1. Quantitative detection of 24 cases of normal pregnancy and 44 cases of eclampsia early serum leptin, TNF-alpha and adiponectin expression levels by enzyme-linked immunosorbent assay (ELISA). 2. Yihong - hematoxylin (HE) staining observed under light microscope normal pregnancy, the placenta and placenta of patients with pathological changes of the structure, with a semi-quantitative immunohistochemistry to detect 21 cases of normal pregnancy and 55 cases of eclampsia early placenta leptin, TNF-alpha and adiponectin protein expression. 3. Reverse transcriptase polymerase chain reaction (reverse transcription polymerase chain reaction, RT-PCR) were detected 10 cases of normal full-term pregnancy and 20 examples of preeclampsia patients (10 cases of mild, moderate and severe 10 cases) placental tissue leptin and adiponectin mRNA expression. Results 1. Mild and severe preeclampsia leptin serum levels were significantly higher than in normal pregnant women, severe group adiponectin levels were significantly lower than normal group, and TNF-alpha level was significantly higher than the normal group; severe group leptin and TNF -alpha showed a positive correlation, negative correlation between adiponectin and TNF-alpha into. 2. Pregnancy placental leptin is mainly distributed in nourishing the cell membrane, cytoplasm and nuclear membrane. Preeclampsia leptin protein expression compared with normal pregnancy significantly increased adiponectin protein expression was significantly lower than normal pregnancy, severe preeclampsia group of leptin and TNF-alpha expression was positively correlated. 3. Normal pregnancy placental leptin and adiponectin mRNA expression were 0.564 ± 0.336,1.610 ± 0.528, while the the preeclampsia placental expression of leptin was significantly increased, the mild group was 0.778 ± 0.175, 1.174 severe group ± 0.556; contrast Conversely, adiponectin expression was significantly decreased in mild group was 1.363 ± 0.475, severe group to 0.636 ± 0.218. And as the disease worsens, the rise of the amount of leptin, adiponectin showed a significant downward trend. Conclusion leptin expression increased with preeclampsia is closely related to other fats cytokines such as TNF-alpha and adiponectin and leptin interaction between each other, may be one of the mechanisms involved in the pathogenesis of preeclampsia. The purpose of the second part of the leptin expression in trophoblasts and trophoblastic proliferation and secretion. The study hypoxia on the chorionic villi trophoblast cell line (TEV-1) expression of leptin and adiponectin, leptin and other adipocytokines preeclampsia from the cellular level. 2. Hypoxia, recombinant human leptin protein and TNF-alpha protein on to nourish cell proliferation and secretory function of the impact of in-depth study of its role in regulating trophoblast cell biological characteristics. Method 1. Cobalt dichloride induced the trophoblastic chemical hypoxia, to simulate in vitro hypoxic environment, the use of reverse transcriptase polymerase chain reaction (reverse transcription polymerasechain reaction, RT-PCR) detection of leptin and adiponectin gene expression. 2. Cobalt dichloride, different concentrations of recombinant human leptin solution and TNF-alpha solution intervention trophoblastic growth in vitro, using quantitative PCR leptin gene expression, MTT (Methl thiazolyltetrazolium, MTT) colorimetric determination of lack of oxygen, leptin and TNF-alpha trophoblastic proliferation. Results 1. With the increase in the time of hypoxia, trophoblastic leptin mRNA expression gradually increased, reached the peak at 12h, compared with the normal culture leptin significantly increased, then decreased gradually; with prolonged hypoxia time, trophoblastic adiponectin mRNA expression gradually reduced, compared with the normal culture at 12h adiponectin expression was significantly reduced, then decreased gradually. 2. A final concentration of 5ng/ml leptin and different concentrations of TNF-alpha can significantly promote the nourish cell leptinmRNA expression, the intervention time this effect had no significant effect. 3. TNF-alpha trophoblastic no effect on proliferation hypoxia inhibition of Sertoli cells to proliferate and to suppress the most obvious 24h; leptin intervention nourish the cells after 24 h, and the final concentration of 10ng/ml leptin inhibitory effect is most obvious; . Concluding preeclampsia increased expression of leptin and adiponectin reduce trophoblastic hypoxia is closely related. Hypoxia can induce similar changes in preeclampsia trophoblast cells. In addition, TNF-alpha can promote trophoblast cells secrete leptin, small doses of leptin and play a positive feedback regulation of trophoblastic leptin secretion. The third part of the leptin, TNF-alpha and to study hypoxic hypoxia on trophoblast invasion ability to influence research purposes, recombinant human leptin protein and TNF-alpha protein on the trophoblast cell lines invasion, further study its biological trophoblastic regulation of the learning characteristics and its mechanism in the pathogenesis of preeclampsia. The method of cobalt dichloride, different concentrations of recombinant human leptin the solution and TNF-alpha solution intervention trophoblastic growth in vitro using Transwell invasion in vitro assay hypoxia, leptin and TNF-alpha on trophoblast invasion ability. Results hypoxia inhibit trophoblast invasion and hypoxia 24h inhibition of the most significant; leptin can promote trophoblast invasion, and with the increased concentration of leptin, trophoblast invasion of the more significant; TNF-alpha is also available promote trophoblast invasion, but its role is independent of concentration. Conclusions were cultured in vitro trophoblast invasion is suppressed under hypoxic conditions, leptin and TNF-alpha can promote trophoblast invasion, which is one of the mechanisms they cause preeclampsia. The purpose of the fourth part of the leptin expression in endothelial cells and endothelial cell injury. Study hypoxia in human umbilical vein endothelial cell line (VEC304) leptin expression from the cellular level of leptin in endothelial cell injury and Preeclampsia relationship. 2. Hypoxia, recombinant human leptin protein and TNF-alpha protein on endothelial cell proliferation and secretory function, in-depth study of the role of the placenta-derived toxic factor in endothelial cell injury. Method 1. Cobalt dichloride endothelial cells induced by chemical hypoxia, simulated in vitro hypoxic environment, the use of reverse transcription-polymerase chain reaction (reverse transcription polymerasechain reaction, RT-PCR) detection of leptin gene expression. 2. Cobalt dichloride, different concentrations of recombinant human leptin solution and TNF-alpha solution intervention endothelial cell growth in vitro, using quantitative PCR leptin gene expression, MTT (Methl thiazolyltetrazolium, MTT) colorimetric determination of lack of oxygen, leptin and TNF-alpha on endothelial cell proliferation. Results 1. With the increase of hypoxia, endothelial cells leptin mRNA expression decreased, but still higher than the control group, training 24h the Shique significantly lower than control, then gradually decreased. 2. The final concentration for 5ng/ml leptin and different concentrations of TNF-alpha can significantly promote the expression of endothelial cells leptinmRNA intervention this effect had no significant effect. 3. Hypoxia inhibits endothelial cell proliferation and suppress the most obvious 48h; leptin and TNF-alpha on endothelial cell proliferation were not affected. Conclusion leptin and TNF-alpha can as placenta-derived virulence factors involved in the injury of endothelial cells, which are contact preeclampsia placental perfusion and systemic vascular endothelial cell activation and (or) damage both part of the pathological changes a key factor.

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CLC: > Medicine, health > Obstetrics and Gynaecology > Obstetrics > Pathological pregnancy ( abnormal pregnancy ) > Gestosis > Pregnancy-induced hypertension
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