Effects of increased complete length mRNA expression of long intracellular domain of leptin receptor on glucose oxidation metabolism in skeletal muscle cells / 中华内分泌代谢杂志

ABSTRACT

Objective To observe the effect of leptin on glucose oxidation with increased mRNA expression of complete length long intracellular domain of leptin receptor ( OBRb) in primary cultured skeletal muscle cells. Methods Skeletal muscle cells were isolated from SD sucking rat and cultured in four groups (recombinant transfection, empty plasmid transfection, non-transfection and control).The recombinant with complete length OBRb cDNA or the empty plasmid were introduced into skeletal muscle cells by clonfectin when the cultured cells were 70% conlluency, leptin and regular insulin were applied into wells and incubated for 1 h, then D-[ U-I4C]-glucose was added into wells and incubated for 2 h. The radioactivity of collected 14CO2 was assayed by scintillation. Expression level of OBRb mRNA in transfected cells was assayed by RT-PCR. Results The ratios of OBRb intraceUular domain mRNA and p-actin mRNA in cells transfected with recombinant, transiected with empty plasimd, and non-transfected were 1.22?0.10,0.41?0.08 and 0.49?0.09, respectively.The expression of OBRb intraceUular domain mRNA in cells transfected with recombinants was significantly increased as compared to the other groups of cells (P