Study On The Expression Of Non-syndromic Cleft Lip And Palate Susceptibility Gene TGFβ3、IRF6、MSX1in Embryonic

Objective:1.Use of retinoic acid induced10-day gestation of pregnant rats formed fetuseswith congenital cleft lip and palate.2.Observed expression of susceptibility genes TGFβ3, IRF6, MSX1of NSCL/P(non-syndromic cleft lip with or without cleft palate, NSCL/P) in mouse embryonic lip andpalate during the different periods of the formation, exploring their mechanism in lips cleftpalate and provide some basis for subsequent experiments and clinical trials.Methods:1.Mouse model of retinoic acid (RA)一induced cleft palate was set up,in first10days of pregnancy (Gestation day, GD)of pregnant mice C57BL/6N, the experimental groupaccording to100mg/kg body weight of pregnant mice were fed retinoic acid once time,thecontrol group received an equal volume of vegetable oil.All killed at GD12、 GD13、 GD14、GD15cesarean and take out embryo.2.Detect TGFβ3, IRF6, MSX1gene mRNA and protein expression and distributionin gestation day GD12, GD13days, GD14days, GD15days use real-time quantitative PCR(quantitative real-time polymerase chain reaction, QRT-PCR) technique andimmunohistochemical techniques..Result:1. Expression of mRNA:Expression levels of TGFβ3mRNA, each group comparedwith the GD12days in the experimental group, the other groups were elevated and there weresignificant differences(p<0.05); each group compared with GD12days in the control group,the other groups were elevated and there were significant differences(p<0.05).Experimentalgroup compared with the control group, the experimental group were higher, there aresignificant differences among GD14days(p<0.05).Expression levels of IRF6mRNA, eachgroup compared with the GD12days in the experimental group, the other groups wereelevated and there were significant differences(p<0.05); each group compared with GD12days in the control group,the other groups were elevated and there were significantdifferences(p<0.05).Experimental group compared with the control group, in addition to theexpression levels of GD15days mRNA in the experimental group higher than control group,the other groups were lower than those in the control group, which GD14significantstatistical difference (p<0.05). Expression levels of MSX1mRNA,each group compared withthe GD12days in the experimental group, the other groups were elevated and there weresignificant differences(p<0.05);each group compared with the GD12days in control group, inaddition to expression of GD13days reduced, other groups were increased, but there was nosignificant statistical difference.Experimental group compared with the control group, inaddition to GD15days, the expression levels were higher in the experimental group, whichGD13day, GD14day statistically significant difference (p<0.05).2.Expression of protein:Expression of TGFβ3protein, each group compared with the GD12 days in the experimental group, the other groups were elevated and there were significantdifferences (p<0.05), each group compared with GD12days in the control group,the othergroups were elevated and there were significant differences(p<0.05).Experimental groupcompared with the control group, the experimental group were higher,in addition to GD13days the other groups were statistically significant differences(p<0.05).Expression ofIRF6protein, each group compared with the GD12days in the experimental group, the othergroups were elevated, in addition to GD14days the other groups were statistically significantdifferences(p<0.05),each group compared with GD12days in the control group,the othergroups were elevated, in addition to GD15days the other groups were statistically significantdifferences(p<0.05).Experimental group compared with the control group, in addition to theexpression of GD15days in the experimental group higher than control group, the othergroups were lower than those in the control group,there were significant differences (p<0.05).Expression of MSX1protein,each group compared with the GD12days in the experimentalgroup, the other groups were elevated, in addition to GD15days the other groups werestatistically significant differences (p<0.05),each group compared with GD12days in thecontrol group,in addition to expression of GD14days increased, there was no significantstatistical difference(p>0.05),other groups were reduced, there were significant statisticaldifference(p<0.05).Experimental group compared with the control group, the experimentalgroup were higher,in addition to GD12days the other groups were statistically significantdifferences(p<0.05)Conclusions:1TGFβ3mRNA and protein expression levels increase, which may related toform lip and palate in the embryonic.2.IRF6gene expression low levels of mRNA and protein inGD12~GD14days, abnormalregulation of palatal growth and development, which led to the occurrence of cleft lip andpalate.3.MSX1gene expression growth levels of mRNA and protein, abnormal regulation of thedevelopment of palatal, leading to the occurrence of cleft palate...