The Research Of The Expression And The Relationship Between Insulin Like Growth Factor â…¡ And Matrix Metalloproteinase 9 In Pre-eclampsia.

PartⅠThe Research of Changes of Insulin-Like Growth Factor-ⅡandMatrix Metalloproteinase-9 in Maternal Serum and Placenta andPathogenesis of Patients with Pre-eclampsia.Objects The purpose of the present study was to investigate the expression ofinsulin-like growth factorⅡ(IGF-Ⅱ)and matrix metalloproteinase 9(MMP-9)in theplacenta and the serum of women with pre-eclampsia(PE)and discuss their role in thepathogenesis of pre-eclampsia.Methods Select 30 examples of pre-eclampsia in pregnant women as the case group(20 examples as mild pre-eclampsia group and 10 examples as severe pre-eclampsia group),15 examples of normal pregnant women giving birth in the same period as the controlgroup.Reverse transcript polymerase chain reaction(RT-PCR)was used to detect theexpression of IGF-ⅡmRNA and MMP-9mRNA in placental tissue on each group;Westernblot analysis was applied to detect the expression of IGF-Ⅱand MMP-9;Immunohis-tochemistry was used to detect the location and expression of IGF-Ⅱand MMP- 9 inplacental tissue and HMIAS-2000 was used to determine the quantity of them;Enzyme linked immunosorbent assay was used to detect the concentration of IGF-Ⅱand MMP-9 inserum of each group.Results①The expressive quantity of IGF-ⅡmRNA and MMP-9mRNA were bothsignificantly lower in women with pre-eclampsia than in the normal group(P＜0.05,P＜0.05).②Western blot analysis showed that both groups express 6.7kDa IGF-Ⅱand92kDa MMP-9 but the levels were significantly lower in women with pre-eclampsia than inthe normal group(P＜0.05,P＜0.05).③Immunohistochemistry analysis tested that the levelsof IGF-Ⅱand MMP-9 were both significantly lower in women with pre-eclampsia than inthe normal group(P＜0.05,P＜0.05).④Enzyme linked immunosorbent assay showed thatthe levels of IGF-Ⅱand MMP-9 were both lower in women with pre-eclampsia than in thenormal group(P＜0.05,P＜0.05).Conclusion The levels of IGF-Ⅱand MMP-9 expression in the placenta tissues andserum of women with pre-eclampsia were significantly decreased and there was a positivecorrelation between them,which indicating that IGF-Ⅱand MMP-9 may play importantrole in the pathogenesis of pre-eclampsia.PartⅡThe Study on Expressions of Insulin-Like Growth Factor-ⅡandMatrix Metalloproteinase-9 in Placental Trophoblast Cells and TheirMutual Relationship in Patients with Pre-eclampsia.Objective By measuring the changes of expression of insulin-like growth factorⅡand matrix metalloproteinase 9 in the placental trophoblast cells of women withpre-eclampsia and normal pregnant women and the changes of levels of matrixmetalloproteinase 9 under the influence of IGF-Ⅱwith different times and differentconcentrations to discuss the role of them in pre-eclampsia and the influence of insulin-like growth factorⅡon the activity of matrix metalloproteinase 9 in the human placentaltrophoblast of pre-eclampsia.Methods Primary cultured 30 examples of placental trophoblast cells of pregnantwomen with pre-eclampsia as pre-eclampsia group(20 examples as mild pre-eclampsiagroup and 10 examples as severe pre-eclampsia group),15 examples of normal pregnantwomen giving birth in the same period as the control group.RT-PCR was used to detect theexpressions of IGF-ⅡmRNA and MMP-9mRNA in placental trophoblast cells of eachgroup and to investigate the levels of MMP-9 in each group with the affection of IGF-Ⅱatdifferent times and different concentrations.Results①The expressive quantity of IGF-ⅡmRNA and MMP-9mRNA were bothsignificantly lower in the placental trophoblast cells of women with pre-eclampsia than inthe normal group and there was a positive correlation between them(r=0.467,P＜0.05,P＜0.05).②Western blot analysis showed that both groups express 6.7kDa IGF-Ⅱand92kDa MMP-9 but the levels were significantly lower in the placental trophoblast cells ofwomen with pre-eclampsia than in the normal group and there was a positive correlationbetween them(r=0.577,P＜0.05,P＜0.05).③After the affection of IGF-Ⅱwith differenttimes and different concentrations,the levels of MMP-9 of each group were increased,ofwitch the pre-eclampsia group rose more obviously and it was showed with time-and dosedependent manner.Conclusion The expression of IGF-Ⅱand MMP-9 in placental trophoblast cells ofwomen with pre-eclampsia were significantly lower than normal pregnant women,suggesting that both of them are involved in the pathological process of pre-eclampsia.IGF-Ⅱcan increase the expression of MMP-9 in trophoblast cells,suggesting that thedecrease of IGF-Ⅱprobably inhibit the expression of MMP-9 in trophoblast cells witchmay be an important factor in the pathogenesis of pre-eclampsia. PartⅢThe Expression of Insulin-Like Growth Factor Binding Protein1 in Pre-eclampsia and Its Affection on IGF-Ⅱand MMP-9 Levels.Objective By measuring the levels of insulin-like growth factor binding protein-1 inthe placental trophoblast cells of women with pre-eclampsia and normal pregnant women todiscuss the role of it in pre-eclampsia and the influence of insulin-like binding protein-1 onthe expressions of IGF-Ⅱand MMP-9 in human placental trophoblast cells.Methods Primary cultured 30 examples of placental trophoblast cells of pregnantwomen with pre-eclampsia as pre-eclampsia group(20 examples as mild pre-eclampsiagroup and 10 examples as severe pre-eclampsia group),15 examples of normal pregnantwomen giving birth in the same period as the control group.Using RT-PCR assay in eachgroup to detect the expressions of IGFBP-1 in placental trophoblast cells and to investigatethe different levels of IGF-Ⅱand MMP-9 with the affection of IGFBP-1 at different timeand different concentrations.Results(①The expressive quantity of IGFBP-1 mRNA was significantly higher in theplacental trophoblast cells of women with pre-eclampsia than in the normal group(P＜0.05).②After the affection of IGFBP-1 with different times and different concentrations,thelevels of IGF-Ⅱand MMP-9 of each group were all decreased,of witch the pre-eclampsiagroup declined more obviously and it was showed with time and dose-dependent manner.Conclusion IGFBP-1 inhibited the expression of IGF-Ⅱand MMP-9 in humantrophoblast cells,suggesting that IGFBP-1 reducing IGF-Ⅱmay be involved in thepathological process of pre-eclampsia.With IGF-Ⅱenhanced MMP-9 gene expression willbe a new train of thought witch may be a prevention and treatment of pre-eclampsia.