Preliminary Study On The Mechanism Of Fat Deposition In Diabetic Rabbits With Syphilis

Objective: In order to explore the renal tissue of diabetic rabbit fatty degeneration mechanism of syphilis.Abnormal adipose metabolism gene candidates in renal tissue of were screened by gene chip technique.Gene expression and pathological change were further detected.Methods: 1.Animal model construction: 29 male New Zealand rabbits were random Ly divided into 4 groups,namely normal control group(C group)5 rabbits,diabetic group(DM group)8 rabbits,syphilis group(TP group)8 rabbits,Diabetes mellitus with syphilis group(H group)8 rabbits.C group(control group),DM group were induced by injection of alloxan 100mg/kg,TP group of rabbit testis inoculation of Treponema pallidum(Treponema Pallidum,TP),H group was injected with four alloxan to induce diabetes and then inoculated with TP infection.Successful standards of diabetes model is fasting blood glucose(> 16.0mmol/L),Criteria of syphilis modeling is TPPA positive.2.biochemical indicators: Blood sample were collected every week,and urea nitrogen(BUN)and creatinine(Cr)change were detected.3.gene chip and m RNA expression level detection: Abnormal adipose metabolism gene candidates in renal tissue were screened by gene chip technique.And m RNA expression level were detected by real-time quantitative PCR,4.WB detection: Total protein of renal tissue were extracted and the protein expression level of ACOX1?ACADM?SCP2 were detected by western-blot technique.5.Pathological changes detection: pathological changes and m RNA expression of ACOX1?ACADM?SCP2 in renal tissue were detected by Oil red O staining?HE staining and immunohistochemistry.Results: 1.In the DM group,6 rabbits were successful,and the average blood glucose level of each rabbit was higher than 16.0mmol/L;In T group P,7 rabbits were successfully established,and the positive staining of TP and TPPA were all positive;5 rabbits of H rabbit model were successfully established.2.With the progress of the disease,compared with the C group,TP group,DM group,H group showed a rising trend of urea nitrogen and creatinine values.Compared with the DM group and TP group,H group was significantly higher.it revealed that with the development of the disease,the renal damage in the experimental group was more serious,compared with T group and DM group(P<0.05),H group are statistical significance.3.the gene chip results showed that ACOX1 gene and ACADM gene expression of TP group was 6.5 times higher than H group,SCP2 gene expression of TP group was 5.4 times higher than H group;ACOX1,ACADM gene expression of C group was 7.4 times higher than H group,SCP2 gene expression in C group was 6.9 times higher than H group;ACOX1 gene expression in C group is 2.5 times more than that of DM group,SCP2 gene expression in C group was 1.8 times higher than DM group.3.m RNA expression of ACOX1,ACADM,SCP2 gene: real-time quantitative PCR results showed that,compared with the normal group,the expression of ACOX1,ACADM,SCP2 genes in TP,DM and H group were down regulated.Compared with TP group and DM group,H group showed lowest expression level(P<0.05).The results are consistent with the results of gene chips.4.WB detection: In renal tissue,the expression of ACOX1,ACADM,SCP2 protein of TP group,DM group,and H group were significantly less than C group.Protein expression level of C group was highest among four groups,followed by the TP group,DM group,H group and the lowest expression,ACOX1,ACADM,SCP2 and m RNA showed the same trend of real-time quantitative PCR results.5.Oil red O staining and immunohistochemistry: rabbit kidney of normal control group showed occasionally lipid droplet coloring.TP group rabbit renal tubular,glomerular had small amount of lipid droplets coloring.DM group,H group of rabbit renal proximal tubule epithelial cells showed obvious lipid droplet deposition.Compared with DM group,H group showed more dense lipid droplets.Immunohistochemical results showed that ACOX1 mainly expressed in the cytoplasm;ACADM is mainly expressed in the cytoplasm and mitochondria;SCP2 is mainly expressed in the nucleus and mitochondria.In normal control group,ACOX1,ACADM,SCP2 showed a large number of positive cells in renal tubules.Compared with the normal control group,the positive cells of TP group,DM group,and H group were significantly reduced.The positive cells of TP group were less than the normal control group,but more than the DM group and H group;followed by the DM group.There are least positive cells in H group.Conclusion: 1.Compared with the Simple disease group,The diabetes mellitus combined with syphilis disease have more severe injury in kidney.2.Compared with the Simple disease group,The diabetes mellitus combined with syphilis disease have more severe fat depositionin kidney.3.Down-regulation of ACOX1,ACADM,and SCP2 expression may be associated with fat deposition in diabetic patients with syphilis.