RESUMO
Objective To investigate the effects of dihydropteridine reductase (QDPR) on regulating apoptosis induced by plamitic acid(PA). Methods The transfection of HEK293T cells experiment was divided into 3 groups. A group was the control vector group. B group was the control vector group induced by PA. C group was the recombinant plasmid QDPR group induced by PA. First, control vector and recombinant plasmid QDPR was respectively transfected into HEK293T cells. After 24 h, PA with concentration of 0. 5 mmol/L was added into the medium of above cells. The cells of control vector group, the cells of control vector group induced by PA and the cells of recombinant plasmid QDPR group induced by PA were cultured for another 24 hours. At last, cells were harvested to detect tetrahydrobiopterin (BH4) and reactive oxygen species(ROS) generation, Beclin 1, Caspase 3 and Beclin 2 expression. Results ① After transfection, the recombinant plasmid QDPR was successfully constructed and expressed in cells.② There was no significant difference between A group and B group in BH4 generation. Compared with B group, BH4 generation increased in C group (P < 0. 05).③ ROS generation was increased in B group compared with A group, and decreased ROS generation in C group compared with B group (P < 0. 05).④ Western blot analysis revealed that Beclin 1 and Caspase 3 increased (P < 0. 05 ) while Beclin 2 decreased in B group compared with A group (P < 0. 05). Compared to B group, Beclin 1 and Caspase 3 decreased while Beclin 2 increased in C group (P < 0. 05 ). Conclusion QDPR may regulate apoptosis induced by fatty acids by decreasing the generation of ROS and increasing the level of BH4 and the expression of Beclin 2 associated with anti-apoptosis.
RESUMO
Objective To detect the expression change of ETFβin diabetic nephropathy rats and study the role of ETFβin fatty acid-induced apoptosis in renal tubules.Methods Diabetic nephropathy model was established by intraperitoneal injection of streptozotocin and unilateral nephrectomy.In vivo ETFβexpression was detected in renal cortex, as well as tubular injury evaluated.In vitro fatty acid-induced apoptosis in renal tubular cells NRK 52E model was established and ETFβrecombinant plasmid was constructed to be transfected into NRK 52E cells and furtherly to observe the effect of ETFβover-expression on the fatty acid-induced apoptosis.Results In the rats model of diabetic nephropathy induced by streptozotocin injection and unilateral nephrectomy, ETFβmRNA and protein expression were decreased as obvious tubular damage occurred.Fatty acids could induce apoptosis in NRK 52E, and ETFβover-expression reduced the apoptosis.Conclusion The expression of ETFβis decreased in diabetic nephropathy model , and ETFβover-expression can reduce apoptosis induced by fatty acid in renal tubular cells.