4 group than the CsA group: VH, 109��6%; VH+K0.2, 101��6%; VH+K0.4, 93��3%; CsA, 70��1%; CsA+K0.2, 73��5%; CsA+K0.4, 83��5; CsA vs. CsA+K0.4, P<0.05). Next, we examined the expression of these markers in islet-specific areas using double immunolabeling for insulin (red fluorescence) and the apoptosis markers selleck chemicals llc (stained with DAB) in the same section (Figure 4D). Each panel in Figure 4D is representative islet and graph of about 20 islets visualized from 10 animals (Bcl-2: VH, 0.038��0.003 mm2; VH+K0.2, 0.030��0.004 mm2; VH+K0.4, 0.032��0.003 mm2; CsA, 0.010��0.002 mm2; CsA+K0.2, ��0.003 mm2; CsA+K0.4, 0.027��0.005 mm2; CsA vs. CsA+K0.4, P<0.05; Bax: VH, 0.008��0.001 mm2; VH+K0.2, 0.009��0.002 mm2; VH+K0.4, 0.010��0.001 mm2; CsA, 0.028��0.003 mm2; CsA+K0.2, 0.021��0.003 mm2; CsA+K0.4, 0.018��0.

002 mm2; CsA vs. CsA+K0.4, P<0.05; active caspase-3: VH, 0.004��0.001 mm2; VH+K0.2, 0.004��0.001 mm2; VH+K0.4, 0.004��0.001 mm2; CsA, 0.018��0.002 mm2; CsA+K0.2, 0.011��0.001 mm2; CsA+K0.4, 0.008��0.001 mm2; CsA vs. CsA+K0.2 or CsA+K0.4, P<0.05). As in the whole pancreatic tissues, immunoreactivity of Bcl-2 was decreased in the CsA group, but this decrease was recovered by KRG cotreatment. Bax and active caspase-3 levels decreased in the CsA plus KRG-treated groups compared with the CsA group. This evidence suggests that KRG might protect pancreatic �� cells from apoptotic cell death during CsA-induced pancreatic injury. Figure 4 Effect of KRG on apoptotic cell death in CsA-induced pancreatic injury. Effects of KRG on Oxidative Stress in CsA-induced Pancreatic Injury Oxidative stress is a major cause of chronic CsA toxicity.

It results in structural and functional impairment of the kidney and pancreas because CsA produces free radical species in the tissues [8], [16], [19], [20]. To investigate the mechanisms underlying the protective effect of KRG against chronic CsA toxicity, oxidative stress was evaluated by measuring 8-OHdG, which is a reliable marker of cellular oxidative DNA damage. Figure 5A shows that the nuclear 8-OHdG immunoreactivity appeared in the pancreas �� cells expressing insulin (red fluorescence). CsA treatment increased the expression of 8-OHdG compared with the VH group, but this increase was attenuated by cotreatment with KRG (Figure 5B: VH, 0.004��0.001 mm2; VH+K0.2, 0.005��0.001 mm2; VH+K0.4, 0.003��0.001 mm2; CsA, 0.017��0.001 mm2; CsA+K0.

2, 0.014��0.001 mm2; CsA+K0.4, 0.010��0.001 mm2; CsA vs. CsA+K0.4, P 0.05). CsA treatment also significantly increased the serum level of 8-OHdG compared with the VH group, but cotreatment with KRG attenuated this increase (Figure 5C: VH, 0.42��0.01 ng/mL; VH+K0.2, 0.42��0.02 ng/mL; VH+K0.4, 0.39��0.01 ng/mL; CsA, 0.57��0.04 ng/mL; CsA+K0.2, 0.52��0.03 ng/mL; CsA+K0.4, 0.42��0.03 Anacetrapib ng/mL; CsA vs. CsA+K0.4, P<0.05).