The Cultivation Of Bama Mini-pigs For Hyperglycemia

With the development of society,the incidence of diabetes is increasing year by year affected by poor dietary habits.Diabetes has become a disease which seriously affects human health after tumor and cardiovascular disease.The insulin peptide amino acid composition of mininature pigs is the most similar to the human,and pigs’characteristics of glucose metabolism and lipid metabolism are very similar to that of human,so the pig is an ideal animal for the study of diabetes and the complications.N-ethyl-N-nitrosourea,a strong gene mutagen,can induce a large number of random mutant phenotypes in a relatively short period of time.The mutant animals with special phenotype could be obtainedby phenotypic selection and genetic tests.The application of ENU can help to establish new models for study of human genetic diseasesand identify the gene function to some extent.Bamamini-pigs were used as the experimental objects in this research,and large mammalian gene mutation models and animal models of human diseases would be screened and established through mutagen ENU.In the screening process,fasting blood-glucose of breeding boars were monitored regularly,and the hyperglycemic breeding boars would be picked out.A stable hyperglycemic family was established by the way of inbreeding and several generations of purification.The main research contents of this paper:(1)The mutagen ENU with the dose of 85mg/kg was injected directly in scrotum site of 12 G0 Bamamini-pigs breeding boars,after which 6 Bama mini-pigs boars that can provide mutagenic semen.Now,a 2907 family with inherited high blood glucose has been established,among which 2907 breeding boar keeps fasting blood glucose at the level of 6.3mmol/L for 25 months(normal fasting blood glucose of Bama mini-pig is 2.0-3.5mmol/L).The average blood sugar for six consecutive months of G3 and G4 were 5.56mmol/L and 6.59 mmol/L.The average blood sugar was 7.48 mmol/L in 3-month-old G5.Intravenous glucose test showed that 2907 and G5 generation met the criteria for diabetes mellitus.(2)The detection results of physiological and biochemical index and related index of blood glucose:there were no statistically significant difference in indexs of WBC,RBC,HGB,HCT,MCV,MCH,MVHC,CHOL,LDL-CH,ALT,TBIL,TP,ALB,GLB and body weight between the two groups in 12-month-old G3(p>0.05).The difference between two groups in PLT,TG,HDL-CH and INS were statistically significant(p<0.05)and the average values of these indexes of the hyperglycemia group were higher than those of normal group.The indexs of WBC,HGB,HCT,MCV,MCH,MVHC,CHOL,HDL-CH,TBIL,TP,GLB,and TG had no statistically significant difference between the two groups in 3-month-old G4(p>0.05),while the diffierence in the indexs of the RBC,PLT,ALT,LDL-CH and ALB were statistically significant between the two groups(p<0.05),and the average values of these indexes of the hyperglycemia group were lower than those of the normal group.The indexs of WBC,RBC,HGB,HCT,MCV,MCH,MVHC,CHOL,LDL-CH,HDL-CH,ALT and GLB had no statistically significant difference between the two groups in 6-month-old G4(p>0.05),while the indexs PLT,TG,TBILand TP had statistically significant difference(p<0.05).The average values of PLT,TG and TP of the hyperglycemia group were lower than those of the normal group,while the TBLT of the hyperglycemia group were higher than that of the normal group.There were statistically significant difference in indexs of TC,HDL-CH,LDL-CH and body weight between the two groups in 3-month-old G5(p<0.05).The average values of INS,FMN and C-Peptide had no statistically significant difference between the two groups in G5(p>0.05).The coagulation function indexs of APTT and PT had statistically significant difference between the two groups in G5(p<0.05).The coagulation function indexs of APTT had statistically significant difference between the two groups in G3(p<0.05).The indexs of APTT and PT had statistically significant difference between the two groups in G5.(3)The liver,pancreas and skin tissue sections of 9 pigs were stained with HE.Microscopic observation showed that the normal group’s islet showed an oval shape and a lot of cells with the glands closely together.The results were observed a transient increase in blood glucose after STZ treatment,insulin structural damage,extreme reduction of islet β cells.The experimental group’s blood glucose was between the normal and STZ groups.The HE observation showed that the islet were not normal.The number of islet β cells also reduced.Islet damage was lower in the STZ group,but the results were consistent with our expectations.The increase blood glucose and pathological changes of pancreas revealed that our design was feasible.(4)We sequenced the total RNA of the liver tissues from three groups of pig(i.e.Normoglycemic,spontaneous hyperglycemic and streptozotocin induced hyperglycemic groups).Comparing to normoglycemic group,EGR1,TNF14 an d AFM had differentially expressed.Pathway analysis with differentially expres sed genes revealed that the most statistically significant metabolic pathways we re related with TNF signaling pathway,Prolactin signaling pathway,PPAR sign aling pathway signaling pathway.(5)The results of the five genes were consistent with the expression of the nine results of the sequencing.ADH4,RHCG and E22200 genes had same trend in changes between the two groups.In general,a 2907 family with inherited high blood glucose has been established,among which result in metabolic disorder,especially in glucose and lipid metabolism and Islet Cells.