| Objective To investigate the relationship between follistatin-like protein3(fstl-3),hypoxia inducible factor-1 alpha(hif-1α)and the onset of early-onset preeclampsia(EO-PE)in the serum of pregnant and late gestational placenta of SD rats.Methods SD rats on the 4th day of gestation were randomly divided into the experimental group and the control group.The experimental group was intraperitoneally injected with ng-nitroarginine methyl ester hydrochloride(l-name)at a dose of 250mg/kg/d for 10 days,and the control group was intraperitoneally injected with normal saline(the same volume as l-name)for 10 days.Serum of tail vein was collected before pregnancy,6th day of gestation(early gestation),13 th day of gestation(second gestation),and 19 th day of gestation(late gestation)of the two groups of rats respectively,and systolic blood pressure(SBP)of the tail arteries of the two groups of rats were measured at the same time.The urine of the two groups of rats was collected on the 18 th day of pregnancy,and the urine protein quantitation was measured on the same day.On the19 th day of pregnancy,the rats in the two groups were anesthetized and underwent caesarean.Placental tissue was weighed for backup,and the babies were weighed respectively.The placental tissues were divided into3 groups,one group was soaked with 4% formaldehyde,the other two groups were homogenized and centrifuged,and the supernatant was taken for reserve.Enzyme-Linked Immuno Sorbent Assay(ELISA)was used to determine the concentrations of fstl-3 and hif-1αin serum and placental homogenate.Immunohistochemical method was used to locate the protein expression sites of fstl-3 and hif-1αin the placenta.A image analysis software named Image pro plus was used to measure the mean Integrated Optical Density(mean IOD[1])of fstl-3 and hif-1αprotein in the placenta,and semi-quantified the expression levels of the above two proteins in the placenta.The expression of fstl-3 and hif-1αm RNA in placental homogenate was determined by Reverse Transcription-Polymerase Chain Reaction(rt-pcr).The experimental data are analyzed statistically.Results From the 6th day of gestation,SBP of tail artery of rats in the experimental group gradually increased,and SBP was measured at the13 th day of gestation,135.80± 5.89 mmhg in the experimental group and108.20± 4.76 mmhg in the control group,the difference value was >15mm Hg,P < 0.05,the difference was statistically significant.On the18 th day of pregnancy,the urine protein quantitation was(4.28±0.54)mg/d in the experimental group and(0.45±0.12)mg/d in the control group,P < 0.05,the difference was statistically significant.Serum fstl-3 concentration was 74.89±4.25 pg/ml in the experi mental group and 78.49±15.26 pg/ml in the control group.Serum concentration of hif-1 alpha before pregnancy in the two groups was 17.18±4.56 pg/ml in the experimental group and 14.87±7.51 pg/ml in the control group.There was no significant difference in serum concen trations of fstl-3 and hif-1alpha in the two groups before pregnancy(P >0.05).On the 6th day of pregnancy,serum fstl-3 concentration(59.60±12.15)pg/ml in the experimental group was lower than that in the control group(88.13±10.63)pg/ml,P < 0.05,showing statistically significant differences.The serum hif-1alpha level of the experimental group was(27.20±3.72)pg/ml,which was higher than that of the control group(13.02±2.84)pg/ml,and the difference was statistically significant(P <0.05).On the 13 th day of gestation,the serum fstl-3 concentration(68.76±8.47)pg/ml in the experimental group increased slightly com pared with that in the control group(93.22±26.55)pg/ml,but the differ ence was not statistically significant(P> 0.05).The serum hif-1alpha level of the experimental group was(26.45±3.83)pg/ml,which was higher than that of the control group(11.04±1.67)pg/ml,and the differ ence was statistically significant(P < 0.05).On the 19 th day of pregnancy,the serum fstl-3 concentration(55.38±14.45)pg/ml in the experimental group was higher than that in the control group(76.44±32.20)pg/ml,with no significant difference(P > 0.05).The concentration of serum hif-1alpha in the experimental group was(24.73±1.27)pg/ml,which was higher than that in the control group(11.26±5.04)pg/ml,and the difference was statistically significant(P < 0.05).The serum concentrations of fstl-3 and hif-1alpha in the control group during each pregnancy and the analysis of variance of design data in the random area group were all P> 0.05,with no statistically significant difference.Serum concentration values of fstl-3 and hif-1alpha in each pregnancy of the experimental group,the analysis of variance of design data in the random area group,P < 0.05,the difference was statistically significant.By bivariate linear analysis,the serum fstl-3 concentration was negatively correlated with SBP,and serum hif-1 alpha serum conce ntration was positively correlated with SBP in the experimental group(Pearson coefficients of bivariate linear correlation analysis were-0.464 and 0.051,respectively.).In the case of similar blood pressure,the higher the urinary protein ration was,the higher the serum fstl-3 and hif-1concentrations were.In the placental homogenate of the experimental group,the concen trations of fstl-3 and hif-1 alpha were(34.75±5.91)pg/ml and(23.01±3.22)pg/ml,respectively,and those of the control group were(10.25±1.99)pg/ml and(2.32±1.30)pg/ml,respectively.In immun ohistochemical sections,mean IOD of fstl-3 protein in the experimental group was(190.44±90.37),hif-1 alpha was(76.87±31.95),and mean IOD of fstl-3 protein in the control group was(8.66±4.37),and hif-1 alpha was(13.8±7.9).Mean IOD expression of fstl-3 and hif-1 alpha in the experimental group was significantly higher than that in the control group(P < 0.05).In the detection of placenta RT-PCR,the relative m RNA expression levels of fstl-3 and hif-1 alpha were(0.73±0.16)and(0.96±0.14)in the experimental group,and(0.16±0.09)and(0.45±0.10)in the control group,respectively.Conclusion(1)In this experiment,serum fstl-3 concentration was negatively correlated with SBP,and serum hif-1 concentration was positi vely correlated with SBP.However,in the case of similar degree of blood pressure elevation,serum fstl-3 and hif-1 concentrations increased with the increase of 24-hour urinary protein volume.(2)Serum levels of fstl-3 and hif-1 may be related to the severity and duration of EO-PE. |
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