| Hexavalent chromium(Cr(Ⅵ))as a heavy metal contaminant has attracted widespread attention in many fields,and many studies have shown that Cr(Ⅵ)can cause liver damage by affecting liver energy metabolism in chickens.Nanoselenium is a new type of low-toxic,strong antioxidant monolithic selenium,which is widely used for heavy metal detoxification.However,it is not clear whether nano-selenium can alleviate the abnormal liver glucose metabolism caused by Cr(Ⅵ)in broiler chickens.The present study aimed to elucidate the potential mechanism of nanoselenium to alleviate the abnormal liver glucose metabolism in broiler chickens caused by Cr(Ⅵ),and provide a test basis for clinical prevention and treatment of Cr(Ⅵ)poisoning in broiler chickens.In vivo experiment: 112 1day old white feather broilers were randomly and equally divided into 4 groups after one week of adaptive feeding: blank control group(C group),Cr(Ⅵ)modeling group(Cr group,37.1 mg/kg K2Cr2O7),nano-selenium group(Se group,0.5 mg/kg nano-selenium),and nano-selenium and Cr(Ⅵ)co-treatment group(Se+Cr group).Seven broilers in each group were weighed,recorded and blood samples collected at 14,21,28 and 35 days of age and then preserved for reserve,and the livers were removed after anesthesia and execution and then preserved for reserve.The effects of selenium nanoparticles on liver damage in Cr(Ⅵ)poisoned broiler chickens were analyzed by observation of histomorphology,pathological sections and biochemical indexes of serum samples and liver tissues;the expression and distribution of mRNA and protein of glucose metabolism related genes(HK2,GLUT1 and PKM2)in liver tissues were detected by RT-qPCR,Western Blot and immunohistochemistry.Cellular assay: Hepatocytes from broiler chicken embryos were isolated and cultured for 14 d for subsequent experiments.The experiment was divided into 4 groups: blank control group(C group),Cr(Ⅵ)treated group(Cr group,16 μmol/L K2Cr2O7),nano-selenium control group(Se group,8 μmol/L nano-selenium),and nano-selenium co-treated with Cr(Ⅵ)(Se+Cr group).After 24 h of drug treatment,RT-qPCR and Western Blot were performed to detect the expression levels of mRNA and protein of glucose metabolism-related genes(HK2,GLUT1 and PKM2)in hepatocytes,In vivo test results: pathological sections showed normal liver tissue structure in C and Se groups,and liver tissue damage in Cr group,and there was a trend of reducing the degree of liver tissue damage in Se+Cr group compared with Cr group.The serum biochemical results showed that the levels of ALT,ASL and GGT were significantly higher in the Cr group compared with the C group(P<0.05);the levels of ALT,ASL and GGT were significantly lower in the Se+Cr group compared with the Cr group(P<0.05);the antioxidant indexes of liver tissues showed that the levels of GSH-PX and SOD were significantly lower in the Cr group compared with the C group(P<0.05)and The antioxidant indexes of liver tissues showed that the GSH-PX and SOD contents were significantly lower(P<0.05)and MDA contents were significantly higher(P<0.05)in the Cr group compared with the C group,and the GSH-PX and SOD contents were significantly higher(P<0.05)and MDA contents were significantly lower(P<0.05)in the Se+Cr group compared with the Cr group.RT-qPCR results showed that the mRNA expression levels of HK2,GLUT1 and PKM2 genes were significantly lower(P< 0.05),and the mRNA expression levels of HK2 and GLUT1 genes were significantly increased in the Se group compared with the Cr group(P<0.05).western Blot results showed that the protein expression levels of HK2,GLUT1 and PKM2 genes were significantly decreased in the Cr group compared with the C group(P<0.05),and the protein expression levels of HK2 and GLUT1 were significantly decreased in the Se+Cr group(P<0.05).The expression levels of HK2 and GLUT1 were significantly decreased in the Se+Cr group(P<0.05);compared with the Cr group,the expression levels of HK2 protein were significantly increased in the Se group(P<0.05).Immunohistochemical results showed that the expression of GLUT1 gene was significantly lower in the group and Se+Cr group compared with the C group;the expression of PKM2 gene was significantly higher in the Se and C groups compared with the Cr group.Cellular assay results: RT-qPCR results showed that the expression levels of mRNA of HK2,GLUT1 and PKM2 genes were significantly lower in the Cr group compared with the C group(P<0.01);the expression of mRNA expression levels of PKM2,HK2 and GLUT1 genes were significantly higher in the Se group compared with the Cr group(P<0.01),and the Se+Cr group had significantly higher expression of PKM2 and Western Blot results showed that the protein expression levels of HK2,GLUT1 and PKM2 genes were significantly decreased in the Cr group compared with the C group(P<0.05),and the protein expression levels of HK2,PKM2 and GLUT1 genes were significantly decreased in the Se+Cr group(P<0.05);compared with the Cr group,the protein expression levels of HK2,GLUT1 and PKM2 genes were significantly higher in the Se group(P<0.05),and the protein expression levels of HK2 genes were significantly higher in the Se+Cr group.In this experiment,using nano-selenium as a supplement for Cr(Ⅵ)-poisoned broiler chickens,the observation of pathological sections,biochemical indexes,liver antioxidant indexes and the expression of mRNA and protein of liver glucose metabolism-related genes HK2,GLUT1 and PKM2 indicated that nano-selenium could regulate the abnormal glucose metabolism in the liver of Cr(Ⅵ)-induced broiler chickens,and thus play a role in the damage in broiler chickens. |
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