Ect of ischemic conditioning on ATP content, reactive oxygen species (ROS), myeloperoxidase (MPO) and NADPH oxidase activity detection from the rat kidney. A reduction in renal ATP content is noted in I15 ?4, I30 ?2 and I60 groups when compared to sham control group. The enhanced ROS (B), MPO (C) and NADPH oxidase activity (D) was significantly increased in the ischemia/reperfusion kidney with different ischemic conditioning treatment. The enhanced renal ROS amounts, MPO activity and NADPH oxidase activity are displayed in an order of I60 > I15 ?4 > I30 ?2 > sham control (C). Data are expressed as mean ?SEM (n = 6 in each group). *, P < 0.05 vs. C group. #, P < 0.05 vs. I60 group.Page 4 of(page number not for citation purposes)Journal of Biomedical Science 2009, 16:http://www.jbiomedsci.com/content/16/1/effect on O2-. levels in the kidney subjected to IR injury. As shown in Figure 2B, the sham control kidney displayed a basal O2-. level (914 ?counts/10 s). In I60 kidneys, increased O2-. activity (3454 ?407 counts/10 PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/27488460 s) was found after 4 hours of reperfusion period. In the I15 ?4 kidneys, the level of O2-. in reperfusion stages were 2898 ?303 ocunts/10 sec. In the I30 ?2 kidneys, the level of O2-. in reperfusion stages were 1897 ?203 ocunts/10 sec. I30 ?2 treatment significantly depressed 40?0 of O2-. level in the IR kidney. The enhanced renal MPO activity (Figure 2C) and NADPH oxidase activity (Figure 2D) are also consistent with the renal ROS data and displayed in an order of I60 > I15 ?4 > I30 ?2 > sham control.In situ localization of 4-HNE formation, tubular autophagy and apoptosis We considered that the high levels of ROS might promote accumulation of renal oxidized products and contribute to autophagy and apoptosis. Renal tubular 4-HNE adducts were pronounced in the renal tubules of IR kidneys in an order of I60 (23.5 ?3.1 ) > I15 ?4 (17.0 ?3.0 ) > 130 ?2 (11.0 ?2.0 ) > sham control kidneys (1.0 ?0.5 ) (Figure 3A ).sion in an order of I30 ?2 > I15 ?4 > I60 kidneys. Bax, Bcl-2, active CPP32 and PARP degradation fragments were sparsely detected in the sham control kidney. I60 markedly increased Bax, CPP32, and PARP expression, but did not affect Bcl-2 expression in the kidney. In contrast, I15 ?4 and I30 ?2 mildly enhanced Bax, Bcl-2, active CPP32, and PARP degradation expression in the kidney. I30 ?2 displayed a less Bax stain than that of I15 ?4 treatment. Beclin-1 and LC3 can be detected in the sham control kidney (Figure 4C). I60, I15 ?4 and I30 ?2 treatment increased LC3 and Beclin-1 expression in the kidneys. However, the expression was implicated in an order of I60 > I15 ?4 > I30 ?2 > sham control.IC ameliorated IR-induced renal Zebularine site dysfunction As shown in Figure 5, after 4 hours of reperfusion, increased BUN and plasma creatinine levels were noted in I60, I15 ?4 and I30 ?2 kidneys. I30 ?2 treatment significantly decreased the values of BUN and creatinine when compared to the I60 group. I30 ?2 was more efficient than I15 ?4 treatment in reduction of BUN and creatinine than after 4 hours of reperfusion. Urine flow dramatically increased from 13.5 ?2.0 L/min (sham control) to 40.2 ?4.0 L/min (I15 ?4 group), 34.1 ?3.2 L/min (I30 ?2 group), and 47.5 ?4.6 L/min (I60 group) after 24 hours of reperfusion. Similarly, urine sodium excretion increased from 7.23 ?1.1 l/min/g in the control kidney to 16.5 ?2.2 l/min/g in I60 kidneys, 14.3 ?2.0 l/min/ g in I15 ?4 kidneys and 12.3 ?1.4 l/min/g in I30 ?2 kidneys. The increased level of the sodi.
