ABSTRACT
AIM:To investigate the effects of seipin gene deficiency on renal injury and the possible mecha-nisms in seipin-/-mice.METHODS:Six-month-old male seipin knockout ( seipin-/-) and wild-type ( WT) mice ( n=8) were used to study 24 h urinary albumin excretion ( UAE) , renal functions, pathological changes, and plasma leptin and adiponectin levels.Seipin mRNA expression in different tissues and each part of the kidney was also measured in WT mice.RESULTS:Real-time PCR analysis showed seipin mRNA expression in WT mice was higher in adipose tissue and testicles, and was also found in the kidney, which was mainly in glomeruli.Compared with control group, seipin-/-group showed increased kidney weight/tibia length (P<0.01), 24 h UAE (P<0.01), creatinine clearance (P<0.01), and glomerular and mesangial surface area (P<0.05).Both plasma leptin (P<0.01) and adiponectin (P<0.05) levels were significantly decreased in seipin-/-mice.CONCLUSION:Seipin gene deficiency in mice leads to renal injury prob-ably by decreasing plasma leptin and adiponectin levels due to lack of adipose tissue.
ABSTRACT
Objective To construct the eukaryotic expression plasmid for the expression of human Connexin26 in COS-7 cells.Methods Total RNA was isolated from human peripheral blood lymphocytes and used as template for the PCR cloning of the human Connexin26 gene.The human Cx26 cDNA containing the 678 bp whole coding region of the human Connexin26 gene was amplified by PCR using specific primers and cloned into the pCI-neo vector to construct the recombinant eukaryotic expression plasmid,pCI-Cx26.The recombinant plasmid was identified by restriction endonuclease digestion,and transfected into COS-7 cells by liposome.The expression of Cx26 mRNA and the protein were analyzed by RT-PCR and SDS-PAGE,respectively.Results Restriction endonuclease digestion analysis verified successful construction of the recombinant plasmid,pCI-Cx26.The expression of Cx26 mRNA and protein in the transfected COS-7 cells were detected by RT-PCR and SDS-PAGE,respectively.Conclusion The eukaryotic expression plasmid for human Cx26 has been constructed successfully with the capability of expression in COS-7 cells.