The Regulation Of Ghrelin To VEGF, Flt-1 In Normal Pregnancy And Pregnancy-induced Hypertension Placental

The development of normal placent was related to the micrangiurn.And the twice recasting vascular physiology were showed inthe normal pregnancy.Pregnancy-induced hypertension(PIH) was existed in the angiogenesis and functional disturbance.VEGF,as one of the vascular growth factor,binded with Flt-1 can regulate the developing of blood vessel in normal and PIH placenta.And the VEGF was regulated by GH,Leptin,NO and ET-1 etc.Ghrelin which can stimulate the GH secretion is a neuroendocrine regulatory factor.It can reform the endodermis function,increase the synthesis and secretion of NOS. And it is the antagon of endothelin,can stimulate the NO synthesis,reform the endothermic function and relax blood vessel.At present,the relationship between ghrelin and VEGF,Flt-1 in the placenta expression;ghrelin expression through the VEGF and Flt-1 regulation;ghrelin regulation on the releasing of NO,NOS;the influence on the placenta blood forming and function regulation are not be reported.The ghrelin,VEGK Flt-1,NO and NOS were detected by the SP-immunochemistry,RQ-PCR and nitrate reductase in the normal pregnancy.Then the ASODN was used to study this relationship in cell level.At last,this relationship was studied in the homopregnant illness and PIH.The results are as follows:1.The expression correction between ghrelin and VEGF,receptor Flt-1 in placenta tissue(1)The protein level of Ghrelin,VEGF and Flt-1 in 6-8 weeks,9-10 weeks,11-15 weeks,16-20 weeks,21-25 weeks,26-30 weeks and 31-38 weeks during the normal pregnancy was detected by the SP immunochemistry.The result showed that the protein of Ghrelin and VEGF,Flt-1 were expressed at the pile and placenta in the normal pregnancy. The positive grana mainly located at langhans cell,pile vascular endothelial cell and the mesenchymocyte around them.The dynamic state changing was shown in different stage in pregnancy.The strength positive expression rate in 9-10w and 16-20w was significant with other stage(P＜0.05).The positive strengthen signal was coincided with the feotus recasting vascular physiology.This showed that the ghrelin was possibly correlated with the protein level of VEGF and Flt-1.(2)The variation of mRNA expression which was in the placenta of 6-8 weeks,9-10 weeks,11-15 weeks,16-20 weeks,21-25 weeks,26-30 weeks and 31-38 weeks during the pregnancy was detected by real-time quantitative PCR.This confirmed that the expression of ghrelin was simple with the VEGF,Flt-1mRNA expression.It was higher in the 9-10w and 16-20w of the pregnancy respective.And these two stages coincided to the placenta recasting vascular physiology.The correlation was verified from the age and room.The ghrelin in placenta regulated the transcription and translation aim gene VEGF and Flt-1, then join in the forming and development of placenta blood vessel.2.The variance regularity of NO releasing and NOS activity in each normal pregnancy stageThe variance regularity of NO releasing and NOS activity in each normal pregnancy stage was studied by detecting,nitrate reductase used,the level of the NO and NOS during the normal pregnancy stage.The result showed that the variance of NO releasing and NOS activity in each normal pregnancy stage was variety in regularity.The three times peaking were showed in the 9-10weeks,16-20weeks and 31-38weeks,the releasing of NO and NOS was correlated with the expression of ghrelin,VEGF and Flt-1.and appeared in the late pregnancy,it showed that the relationship with the childbirth priming and womb smooth muscle loosen.3.Antisense oligonucleotide technology(ASODN)The influence of ghrelin with human umbilical vein endothelial cells(HUVEC),the protein and mRNA level of VEGF and Flt-1,releasing level of NO and NOS was detected by antisense blocking ghrelin transcription.The inhibited ghrelin optimization,using the ASODN concentration and diversity culture time,was 20μmol/L and 36h respetive.The expression of HUVEC and VEGF,Flt-1 was detected.And after antisense blocking ghrelin, the expression level of VEGF and FLT-1 in protein and mRNA descended.And it was significant to the control group and the SCON group(P＜0.05).The NO content and NOS activity were lower(P＜0.05).The expression of VEGF and Flt-1 in mRNA and protein level was lower and the content NO and NOS activity was down after antisense blocking ghrelin in vitro.4.The expression of ghrelin,VEGF,Flt-1,NO and NOS in normal and PIH women was detected by immunochemistry,RQ-PCR and nitrate reductase respective.The mRNA and protein level of VEGF and Flt-1 in PIH was significant to the normal women,the influence of ghrelin in PIH(p＜0.05).The ghrelin can regulate the VEGF and Flt-1 in mRNA and protein level down,and influence the forming and development of the placenta blood net.The NO content and the NOS activity were down in the PIH pregnant women placenta.This showed that the ghrelin regulated the VEGF and influenced the the NO (blood vessel relaxing factor) down.And the placenta angiospasm,blood pressure highten, and the PIH were caused also by this. The ghrelin,VEGF,Flt-1,NO and NOS were detected by the SP-immunochemistry, RQ-PCR,cell culture in vitro,ASODN and nitrate reductase in the normal pregnancy,PIH placenta and the cultured HUVEC antisense blocking ghrelin in vitro.The ghrelin was different with the VEGF,Flt-1,NO and NOS in each normal pregnancy stage.And this was concide to the twice periods of recasting vascular physiology.So it was supposed that ghrelin influenced the forming and development of the pregnancy placenta blood vessel. The VEGF and Flt-1 in mRNA and protein level were down regulated by the antisense inhibiting ghrelin in the HUVEC.It was same to the NO content and the NOS activity. Those factors were down in the pregnancy placenta.The ghrelin played a part role in the forming and development placenta blood vessel in normal pregnancy and the occurrence and development of the PIH.It was provided a novel direction to study the ghrelin in the reproduction and clinical OG disease.