| Objective: This study aimed to explore the biological effect of ZAG in the association between arsenic poisoning and diabetes mellitus,and to provide new ideas for the association and diagnosis and treatment of arsenic exposure and diabetes mellitus.Methods: 40 SPF SDrats were randomly divided into control group,diabetes mellitus group,low,medium and high dose of arsenite group(low arsenic group,middle arsenic group,high arsenic group),each group of 8,male and female half.The arsenic-treated SD rats(oral)arsenic were continuously arsenic for 15 weeks,the dose was 0.45mg/kg,2.25mg/kg and 11.25mg/kg from low to high.Body weight of rats in each group was determined weekly during the experiment.After 5 weeks,alloxan was intraperitoneally injected into the diabetes mellitus group to establish the model of diabetes mellitus SD rat.The rats in each group were tested for fasting blood glucose and urine glucose every two weeks.At the 15 th week,the organs of heart,liver,spleen,lung,kidney in each group were harvested after intraperitoneal anesthesia,and as required the blood samples were centrifuged and stored at-80 ?,after the experiment.COD-PAP enzymatic determination of total cholesterol(TC),triglyceride(TG);direct determination of low-density lipoprotein cholesterol(LDL-C),high-density lipoprotein cholesterol(HDL-C);Enzymatic colorimetric determination of glycated hemoglobin(HAbC1);insulin(INS)and insulin-like growth factor-1(IGF-1)were measured by ELISA.The pathological changes of liver and kidney were observed by HE staining.Determination the arsenic of urinary,blood,liver and kidney of SD rats by graphite furnace atomic absorption spectrometry.The contents of glutathione(GSH),malondialdehyde(MDA)and superoxide dismutase(SOD)in liver tissue were measured by ELISA.Determination of reactive oxygen species(ROS)activity in liver tissue by chemiluminescence method.Quantitative real-time polymerase chain reaction(qRT-PCR)was used to detect the mRNA expression of zinc-?2 glycoprotein(ZAG),peroxisomal proliferators-activated receptor ?-coactivator-1?(PGC-1?),insulin(INS)and insulin receptor substrate-1(IRS-1)in SD rat liver tissue.Western blot was used to detect the protein expression of ZAG,PGC-1? and IRS-1 in rat liver tissue.Results: 1.At the 15 th week of experiment,the body weight of rats in diabetes mellitus group and high arsenic group was lower than that in control group(P<0.05),there was no significant difference in body weight between diabetes mellitus group and diabetes mellitus group(P>0.05).2.Fasting blood glucose levels in the high arsenic group were higher than those in the control group at the 11 th,13th and 15 th weeks of the experiment,and the urine glucose of the high arsenic group was positive at 15 weeks(P<0.05).3.The levels of TC,TG,LDL-C and HAbC1 in diabetic group and each arsenic-treated group were significantly higher than those in control group,INS,IGF-1 and HDL-C were lower than those in control group(P<0.05)There was no significant difference in LDL-C,HDL-C,INS and IGF-1 between rats and diabetic rats(P>0.05).4.The liver and kidney organ coefficients of diabetic rats and high arsenic rats were significantly higher than that of the control rats(P<0.001),but there was no significant difference in the liver organ coefficient between high arsenic rats and diabetic rats(P>0.05).5.In the control group,the structure of liver and kidney were normal,with no obvious pathological changes.The pathological changes of liver and kidney were found in the diabetic group.The arsenic-stained rats became more and more serious with the increase of the dose.6.Arsenic in rats with arsenic,blood arsenic and liver,kidney arsenic levels were significantly higher than the control group(P<0.05).7.The activities of GSH and SODin diabetic rats and arsenic-treated rats decreased in turn,while MDA and ROS increased in turn(P<0.05).There was no significant difference in SODbetween high arsenic group and diabetic group(P>0.05).There was no significant difference in GSH,MDA,SODand ROS between low and middle arsenic group and diabetic group(P>0.05).8.The mRNA and protein expressions of ZAG,PGC-1?,INS and IRS-1 in liver tissues of low,middle and high arsenic groups were decreased with the increase of arsenite concentration(P<0.05).The mRNA expressions of ZAG,PGC-1?,INS and IRS-1 in the arsenic group and diabetic group were not significantly different(P>0.05).There were no significant differences in the expression of ZAG,PGC-1? and IRS-1 in the liver between high arsenic group and diabetic group(P>0.05).Conclusion: Arsenic down-regulated the expression of ZAG by oxidative stress and affect the function of PGC-1?,reduced disorders of glucose and lipid metabolism in hepatic,aggravated the dysfunction of insulin secretion and synthesis,and increased blood glucose,indicated the arsenic may cause occurance of diabetes mellitus. |
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