The Product Of Inflammatory Cytokines In Cultured Human Umbilical Endothelial Cell Induced By Sera Of Preeclampsia And The Inhibition Of Unlinastatin

Objective: To investigate the inhibitory effects of different concentrationof ulinastatin on the inflammatory cytokines to cultured human umbilicalendothelial cell(HUVEC) induced by preeclamptic sera in vitro, to investigatethe inflammatory responses ,increased permeability and the mechanism ofproteinuria in preeclampsia from the view of endothelial cell, provide theevidence of experiment theories for treatment and scientific research.Methods: The cultured humbilical vein endothelial cells (HUVECs) weredivided into 4 groups.①Control group:Normal saline was added into cellculture.②Normal pregnancy group: 15% normol pregnancy sera was addedinto cell culture.③Preelampsia group : 15% preeclamptic sera was added intocell culture.④Ulinastatin group: ulinastatin(50U／L、100U／L、500 U／L、1000 U／L )was added separately into HUVEC culture which preeclamptic serawere added as preeclampsia group. The nitrite content of the culture medium incontrol group, normal pregnancy group, preelampsia group and ulinastatin group was measured by nitrate reductase method；The mRNA expression ofiNOS of HUVEC in control group, normal pregnancy group, preelampsia groupand ulinastatin group was detected by real-time reverse transcription polymerasechain reactions by dsDNA-binding dyes of SYBR Green;the protein of TNF-ɑof HUVEC in control group, normal pregnancy group, preelampsia group andulinastatin group was detected by ELISA.Results: 1. Levels of NO was no significantly statistical between normolpregnancy group and control(P >0.05) .Levels of NO in preeclampsia groupwere significantly high vs control（P <0.001）; Levels of NO in preeclampsiagroup were significantly high vs normol pregnancy group（P<0.001）;Ulinastatingroup（except 50U/L）could significantly inhibitor the production of NOstimulated by preeclampstic sera（P<0.01）.2. The mRNA levels of iNOS was no significantly statistical betweennormol pregnancy group and control(P >0.05) . The mRNA levels of iNOS inpreeclampsia group were significantly high vs contro（lP <0.001）; The mRNAlevels of iNOS in preelampsia group were significantly high vs normolpregnancy group（P<0.001）;Ulinastatin group（except 50U/L）couldsignificantly inhibitor the production of NO stimulated by preeclampstic sera（P<0.05）.3. Levels of TNF-ɑwas no significantly statistical between normolpregnancy group and control(P >0.05) .Levels of TNF-ɑin preeclampsiagroup were significantly high vs control（P <0.001）; Levels of TNF-ɑin preelampsia group were significantly high vs normol pregnancy group（P<0.001）;Ulinastatin group（except 50U/L）could significantly inhibitorthe production of TNF-ɑstimulated by preeclampstic sera（P<0.001）.Conclusion: Preclamsia serum can activate vascular endothelial cells torelease inflammatory factors NO and TNF-alpha is one of the importantmechanism of the excessive inflammatory response in preeclampsia;a certainlevel of ulinastatin inhibit the inflammatory factors released by preeclampsiavascular endothelial cells and may be considered as anti-inflammatoryendothelial cells protective agent used in clinical .