Alteration And Significance Of The Differently Expressed Protein In Serum From Patients With Hypertensive Disorder Complicating Pregnancy At Mid Trimester

Objective:To explore the serum proteomic fingerprints of Patients with hypertensive disorder complicating pregnancy at mid trimester by use of surface enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS), to screen biomarkers of the differently expressed proteins in the different groups of hypertensive disorder complicating pregnancy patients, to search non-invasive markers that would be useful and specific for early diagnosis,so as to elucidate the pathogenic mechanism of hypertensive disorder complicating pregnancy.Method:Total Serum samples collected from 39 hypertensive disorder complicating pregnancy patients and 30 normal pregnant women at 15～20 gestation week。All Serum samples were detected by surface enhanced laser desorption/ionization time-of-flight mass spectrometry(SELDI-TOF-MS) using mobilized metal affinity capture (IMAC3-Gu) proteinchip arrays. Differences of mass spectrums between hypertensive disorder complicating pregnancy patients and healthy women were analyzed by Biomarker Wizard software.The we used learn-group including 36 of 69 to set up diagnostic patterm. Some different protein peaks were picked out and a classification tree was developed for identification of hypertensive disorder complicating pregnancy patients using Biomarker Patterns software.The classification model was validated by an independent sample from the test-group.The differential proteins were identified by searching the UniPort protein database according to their molecular weight and verified by enzyme-linked immunosorbent assay (ELISA), Also try to probe the valuation in the prediction of gestation hypertension disease in the pregnant medium-term. Result:1. In the rang of M/Z from 1500 to 20000,Six peaks were significantly different between Gestational hypertension patients and normal pregnant women.In the serum of Gestational hypertension group,the relative amount of protein which M/Z are 8913,11050 and 11687 is obviously higher than normal group (P< 0.01), the relative amount of proteins which M/Z are 1539,5333 and 5801 is obviously lower than normal group (P< 0.01)。2. In the rang of M/Z from 1500 to 20000, Eight peaks were significantly different between preeclampsia patients and normal pregnant women.. In the serum of preeclampsia group,the relative amount of protein which M/Z are 11050 and 11687 is obviously higher than normal group (P< 0.01), the relative amount of proteins which M/Z are 1539,3684,5333,5900,7748 and 9262 is obviously lower than normal group (P< 0.01)。3. The classification diagnostic tree built by BPS consists of 4 peaks which M/Z is 11050 Da,1539Da,2795 Da and 5063 Da. It can distinguish different groups with sensitivity and specificity of all 100% in training mode. In validation test, the sensitivity specificity and precision rate were 75%,94.12%,84.85%, respectively.4. The proteins which M/Z are 1539,5333,11050 and 11687 are the special biomarkers in serum of hypertensive disorder complicating pregnancy. Database search revealed that the peak at a m/z of1539,11050 and 11687 corresponded to Fibrinopeptide A (FPA), C-C motif chemokine 2(MCP-1), Serum amyloid A protein (SAA).one protein peak which M/Z is 5333 may be a uncharacterized protein.5. A significant positive correlation was observed between the relative amount of protein which M/Z is 11050 and serum MCP-1 level of the hypertensive disorder complicating pregnancy group (r=0.928, P<0.01).6. ELISA detection of MCP-1 showed that serum MCP-1 level was significantly higher in hypertensive disorder complicating pregnancy patients than in healthy controls (P< 0.05). Maternal serum MCP-1 levels had predictive value at 15-20 gestational weeks by ROC curve, the area under the curve was 0.774 for HDCP and 0.909 for preeclampsia, the best cut-off values for HDCP and preeclampsia of MCP-1 was 127.075 pg/ml, the sensitivity, specificity, positive predictive value, negative predictive value and likelihood radio were 64.10%,80.00%,51.02%,87.27% and 3.21 for HDCP, respectively; the sensitivity, specificity, positive predictive value, negative predictive value and likelihood radio were 94.40%,80.00%,41.46%, 98.97% and 4.72 for preeclampsia, respectively.Conclusion:1. The differently expressed protein in serum screened by SELDI-TOF MS could act as the biomarkers of the pregnant hypertension's occurence and development,it can also lay a foundation for the research in protein of the pregnant hypertension pathogenesis.2. Compared to normal pregnant group,there are some different proteins in serum of Gestational hypertension group and preeclampsia group,which indicated that Gestational hypertension has different pathophysiological changes with preeclampsia.3. Search the database to identification of the 3 kinds of the differently expressed proteins. The result of searching database shows that the development of HDCP may correlate to dysfunction of blood coagulation, Immune and inflammatory reaction, oxidative stress.4. The diagnostic patterm built in this study maybe useful for diagnosis of HDCP.5. The level of MCP-1 in HDCP group showed significantly higher than that of the normal group, Serum MCP-1 may play an important role in the development of hypertensive disorder complicating pregnancy. The level of serum MCP-1 at second trimester may be used as an early predictive marker of preeclampia.