| Objective:Cleft of palate (CP) is one of the most common birth defects in humans, which account most of skull and maxillofacial deformity and brings huge suffering to life and psychology of patients and their families.Presently,there are many scholar have done many basic and clinical research for happen mechanism of cleft palate,and they have gained many success.But the happen mechanism of cleft palate is not clear,because it is a disease of multifactorial inheritance.The research for happen mechanism of cleft palate is very helpful for obviate and antepartum diagnose of cleft palate.Retinoic acid (RA)to build an important effect on embryonic development,organ formation,cell differentiation and retain homeostasis of biosystem.Excess RA take place many secondary effects,such as make mistake of gene expression to induce anormogenesis of embryo,and induce abnormity of tissue and organ.The function of RA signal is implement through gentic transcriptional control,like growth factor,cytokine and their receptor,hormone,transcription factor,kinase and phosphatase,desmoenzymes and effective factor and so on,they are all adjusted by RA.Recently,many abroad scholar research for relationship of RA controlling gene expression and induce cleft palate.But the mechanism of RA induce cleft palate is not clear now.Transforming growth factorβsuperfamily is a group of cytokine with many biologic activity,and participate embryonic development in many ways,and it can encourage generation,growth differentitate and suppressor cell growth for many different kind cells,it is a kind of two-ways regulation factor for cell growth.Now some scholar extract DNA from sample , choice definite genetic mark primer for PCR,then detect product of PCR.Bolting out that TGFβmaybe a predisposing gene of cleft palate.So,this experiment build C57BL/6J model of cleft palate,detect the TGFβ1 and TGFβ3 space-time expression change on palate of RA induce cleft palate and normal C57BL/6J mouse by immunohistochemistry,prepare for the advanced research.Method:We elect C57BL/6J mouse about 10 weeks of age,female mouse were over 25g, male mouse were over 20g.Housed overnight at 5pm F:M =2:1,checked for vaginal plugs the next morning 8am,which was designated as day 0 of gestation. Totally (10) pregnant females were obtained. All the pregnant mice were randomly divided into two groups: RA-treated group (5)and control group (5).The first group filling 10mg/ml RA vegetable oil solution per 0.01ml/g at 10am on GD10,the other group filling 0.01ml/g vegetable oil.All killed at GD14,GD15,GD16 3 time point and take out cyema.Cyema's head be fixed,bladed after embed,then detect the expression of TGFβ1 and TGFβ3 on mouse palate by immunohisto- chemistry.As a result,we find out the expression of TGFβ1 and TGFβ3 on the palatine process of RA induce cleft palate model and normal mouse.Result:1 The express of TGFβ3 is cut down in mesenchymal of RA affected and control group,the express of TGFβ3 is same at GD14,GD16 in mesenchymal of RA affected and control group,but at GD15 the express of TGFβ3 is higher ofRA affected group;and the express of TGFβ1 is same at GD14,GD15,which step up by the development of embryonic,but atGD16 the express of TGFβ1 is higher in RA affected group.2 The express of TGFβ3 be high straightly at medial edge epithelial of RA affected group,but it cut down in control group by the development of embryonic;the express of TGFβ1 at medial edge epithelial of RA affected group is step up by the development of embryonic obviously,but it is step up firstly,then cut down in control group.3 The express of TGFβ3 at epithelium of nasal side is same at GD14 of RA affected and control group,which higher at GD15 of RA affected group and higher at GD16 of control group; the express of TGFβ1 at epithelium of nasal side is same at GD14 of RA affected and control group too,which lower at GD15 of RA affected group and lower at GD16 of control group obviously.Conclusion:1 At the RA-treated group,the express of TGFβ3 higher in mesenchymal at GD14 and GD15,but the express of TGFβ1 is lower;that TGFβ1and TGFβ3 may cut down the reduplication of mesenchymal,then influence the raise of palatie process,induce cleft of palate.2 The express of TGFβ3 be high straightly at medial edge epithelial of RA affected group,which make the medial edge epithelial dysdifferentiation to epithelium of oral side;then the high express of TGFβ1 at GD16 maybe enhancement the dysdifferentitation of medial edge epithelial,all above may suppose that TGFβ1and TGFβ3 may induce the dysdifferentitation of medial edge epithelial,induce cleft of palate.3 At GD15, the express of TGFβ3 higher at epithelium of nasal side,but the express of TGFβ1 is lower at RA-treaded group,which induce the dysdifferentitation of nasal side epithelium.That may suppose TGFβ1and TGFβ3 may induce the dysdifferentitation of nasal side epithelium,which don't touch with anasl septum,induce cleft of palate.
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