Study On The Distribution And Variation Law Of Glut2 In Pancreas Of Three-way Hybrid Pigs At Different Developmental Stages

Glucose in animals is mainly transported through the family of glucose transporters(Gluts),of which glucose transporter 2(Glut2)is mainly present in liver and pancreas tissues.When the glucose content in the animal’s body is low,Glut2 shows a very low affinity,and when the glucose concentration increases,Glut2’s affinity for glucose increases.Therefore,Glut2 and glucokinase in the animal pancreas work together to maintain the stability of the glucose concentration in the animal by sensing the change in glucose concentration.In 201 0,Atwater developed a method for separating islets based on selective osmotic shock(SOS)based on the characteristics of Glut2.This method can avoid the negative effects of traditional collagenase and has great potential for clinical application.The principle is to use the particularity of Glut2 transporting glucose in the islets to maintain the stable internal and external osmotic pressure of the islets in a high-glucose environment,while the exocrine glands that do not contain Glut2 cause the cells to burst due to excessive intracellular osmotic pressure,thereby achieving islet separation effect.Studies have shown that newborn pigs are not suitable for separating islets by SOS,while young pigs and adult pigs are suitable for separating islets by SOS,but the reason is unknown.Therefore,this study used immunohistochemistry,WB,RT-PCR and other technologies to study the distribution characteristics and changes of Glut2 in porcine pancreas tissues at different development stages,to further improve and establish a new animal islet isolation method.The results are as follows:1.Immunohistochemistry showed that under the light microscope,in the negative control group,no complete islets were seen in the pancreas of newborn pigs.Young pigs,adult pigs,and islets developed well,and the outline of islets was clear,elliptical or round,and none were seen.Positive signal expression.In the experimental group,the newborn pigs showed scattered Glut2 positive signal expression,and the young pigs and adult pigs showed that the Glut2 immunopositive signal was localized on the pancreatic islet cell membrane and was brownish yellow.2.Western blotting results showed that compared with the control group,the relative expression of Glut2 in newborn pigs was 0.26860±0.1216,the relative expression of Glut2 in young pigs was 1.271 8±0.1211,which was 4.75 times that of newborn pigs,and the relative expression of Glut2 in adult pigs was 0.82887.±0.1029,which is 3.08 times that of newborn pigs.The expression of Glut2 in young pigs and adult pigs are significantly higher than that of newborn pigs,and the expression of young pigs is significantly higher than that of adult pigs.3.Real-time PCR results showed that the relative expression of Glut2 mRNA of newborn pigs was 1.000±0.04,the relative expression of young pigs was 0.02919±0.01792,which was 0.03 times that of newborn pigs,and the relative expression of adult pigs was 0.07274,which was that of newborn pigs.0.07 times,which is 2.5 times that of young pigs.The relative expression of Glut2 mRNA in young pigs and adult pigs is significantly lower than that of newborn pigs,and there is no significant difference between the two.4.At 500mmol/L,the output of islets of newborn pigs separated by SOS is 171±9.00 per/g,young pigs are 5840±625 per/g,which is 34 times that of newborn pigs;adult pigs are 6951±356 per/g,which are newborn pigs The amount of islets of young pigs and adult pigs is 41%higher than that of newborn pigs,and there is no significant difference between the two.It can be seen that the newborn pig Glut2 is scattered in the pancreas,and the young pig and the adult pig Glut2 are distributed on the islet cell membrane.The relative expression of newborn pig protein is significantly less than that of young pigs and adult pigs,and young pigs are significantly higher than adult pigs.The relative gene expression of newborn pigs is extremely higher than that of young pigs and adult pigs,and there is no significant difference between young pigs and adult pigs.When the SOS method is used to separate the pancreas and pancreatic islets of pigs of different ages after embryos,the adult pig is the most suitable for the separation by the SOS method,and the number of separation is the largest,which is 6951± 356/g.