| Objective:We observe the expression of ICAM-1 in endothelial cells of preeclampsia patients’(damp heat and blood stasis syndrome) umbilical cord samples;TO study the influence of preeclampsia patients’ sera on proliferation and the expression of ICAM-1 of human umbilical vein endothelial cells(HUVECs) in vitro;To study the effects of Baicalin on the proliferation of serum-induced cells, then select the best concentration;To study the effects of BAI on the expression of ICAM-1 in serum-induced cells. To investigate the targets and possible mechanisms of BAI for the prevention and treatment of preeclampsia,standardize the most effective concentration of Baicalin and provide theoretical and experimental basis in further widely clinical application.Methods:(1)Clinical research,selecting 50 cases of preeclampsia(damp heat and blood stasis syndrome)and 50 cases of normal pregnant women.Blood and serum were separated under sterile conditions, 56℃ 30 min to inactivate complement, packed then stored at-20℃.After delivery, collected umbilical cord and stored at-80℃.Twenty sections of umbilical cord were randomly selectedin each PE and NP group, SABC assay the expression of ICAM-1.(2) Empirical research,Human umbilical vein endothelial cells, referred to as HUVECs,unlimited subculture, easy experimental operation.Establish PE endothelial damage model induced by Preeclamptic serum. HUVECs pretreated with normal pregnant women’s serum as a negative control.Group a(preeclampsia model), group b(normal model as negative control group), group c(PE model + baicalin).HUVECs were harvested from group a and b,we adjust the cell density then transfer them into 96-well plates after digestion with trypsin, five parallel wells were used for each treatment. After attachment, the cells were subjected to the different treatments of drugs.After culturing for 24 hours, 10 ul of cck-8 was added to each well. Adjusting the cell concentration to 2×104cells/well, 1×104 cells /well, 5×103 cells/well, conducting three times of experiments.HUVECs were harvested from group a and group b, according to the results of exploration,then transferred into 96-well plates, adjusted optimal cell density and the level of concentration of baicalin, as above.Collecte the three groups of cells separately, RT-PCR and Western blotting were used to measure the m RNA and protein expression levels of ICAM-1 in HUVECs.Results:(1)Immunohistochemical measured the expression of ICAM-1 in human umbilical vein endothelial cells: preeclampsia umbilical cord(51.42±4.46)was significantly higher than normal pregnancy group(14.68±2.73), p<0.01.(2)Compared with the normal group(1.18±0.07),the activity of endothelial cells of PE model(1.49± 0.08)shows obvious proliferation.Baicalin inhibits the proliferation of HUVECs induced by PE,p<0.05.(3)The expression of ICAM-1 m RNA and protein in HUVECs of PE was significantly higherthan NP, Baicalin could inhibit the expression of ICAM-1 of HUVECs induced by PE serum,p<0.01,but there are no obvious statistical significance between NP and BAI,p>0.05.Conclusions:(1)Clinical research,ICAM-1 shows positive expression in preeclampsia(damp heat and blood stasis syndrome) patients’ umbilical endothelial.(2)Empirical research,preeclamptic serum could stimulate HUVECs’ imbalance of proliferation;Baicalin could inhibit the over-expression of ICAM-1 and the imbalanced proliferation of HUVECs of PE;baicalin in 60μg/ml, effectively inhibited the proliferation of endothelial cells and the activity of cells is closest to the normal group; Baicalin perhaps by inhibiting imbalanced proliferation of vascular endothelial cells under pathological environment and the over-expression of cell adhesion molecules in order to maintain the balance,reduce vascular endothelial adhesion and protect endothelial cells.(3)From the basic experimental studies, 1×104cells/well, cck-8 add 4h,make the most suitable OD value,provide experience for the following experimental study. |
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