Binding buffer (400 ��L) was added to dilute the samples before analysis on a flow cytometer. The cells positively labeled for Annexin V were considered apoptotic. These experiments were conducted three times at least. Statistical Analysis Data are expressed as mean �� standard error of at least three independent ref 3 experiments. Groups were compared using a one-way analysis of variance with Bonferroni��s correction (SPSS, Chicago, IL). Statistical significance was assumed at P<0.05. Results Effect of KRG on Pancreatic Function in CsA-induced Pancreatic Injury Table 1 shows the effects of 4 weeks of CsA and KRG (K) treatment on the basic parameters of the experimental groups. Both CsA+K groups (one treated daily with 0.2 g/kg and one with 0.4 g/kg of KRG) showed lower serum creatinine (Scr) and blood urea nitrogen (BUN) than the CsA only group.

KRG cotreatment did not affect the CsA level in either whole blood or pancreatic tissues, indicating that drug interactions did not occur at these doses. Baseline blood glucose levels did not differ among the four groups (Figure 1A and B). However, the blood glucose level after intraperitoneal glucose loading was significantly higher in the CsA group than in the VH group, but cotreatment with KRG significantly decreased the blood glucose level at 30 and 60 min after glucose loading compared with the CsA group. KRG cotreatment with CsA decreased the calculated area under the curve for glucose (AUCg) compared with the CsA group (Figure 1C: VH, 1229��55 mg/dL/min; VH+K0.2, 1174��49 mg/dL/min; VH+K0.4, 1360��35 mg/dL/min; CsA, 1563��39 mg/dL/min; CsA+K0.

2, 1447��32 mg/dL/min; CsA+K0.4, 1368��44 mg/dL/min; VH vs. CsA, CsA vs. CsA+K0.2 or CsA+K0.4, P<0.05). The fasting insulin level was significantly lower in the CsA group than in the VH group. It was significantly increased in both CsA+K groups compared with the CsA group (Figure 1D: VH, 0.30��0.05 ng/mL; VH+K0.2, 0.33��0.05 ng/mL; VH+K0.4, 0.30��0.04 ng/mL; CsA, 0.15��0.01 ng/mL; CsA+K0.2, 0.19��0.01 ng/mL; CsA+K0.4, 0.30��0.06 ng/mL; VH vs. CsA, CsA vs. CsA+K0.2 or CsA+K0.4, P<0.05). Thus, oral KRG administration during CsA-induced pancreatic dysfunction in mice improved glucose tolerance and restored defective insulin secretion, suggesting that KRG has an antihyperglycemic effect. Figure 1 Effect of KRG on pancreatic function in CsA-induced pancreatic injury. Table 1 Effect of KRG on basic parameters. Effect of KRG on Pancreatic �� Cell Area in CsA-induced Pancreatic Injury Double immunofluorescence for insulin (red fluorescence) and glucagon (green fluorescence) in the VH group and Carfilzomib VH+K groups showed a strong and uniform pattern of staining in a large proportion of the islet �� cells (Figure 2A).