Fig. 8

Western blot analysis showing the expression levels of β-actin, HIF-1α, VEGF, PKM2 and RBP2. Lane 1 or 2 of picture a1 shows a sham retina subjected to the sham procedure (Sham), or a ischemic retina pre-administered with vehicle (Vehicle+I/R), respectively. Lane 3 shows a retina that received ischemia together with pre-ischemia administration of 1 g/kg/day DNL (DNL1.0 + I/R). Lanes 4~ 6 show retinas that underwent ischemia together with pre-ischemia administration of 10 μM/5 μl JIB-04 (RBP2 inhibitor), 4 μM/5 μl shikonin (PKM2 inhibitor), or 125 μg/5 μl avastin (anti-VEGF), respectively. Each bar in picture a2 represented the ratio of RBP2, PKM2, VEGF and HIF-1α to β-actin. ** indicates a significant (P < 0.01) difference between the Sham retina and the ischemic retina pre-administered with vehicle (Vehicle+I/R). † or †† indicated a significant (P < 0.05 or P < 0.01) difference between the “Vehicle+I/R” group and the ischemic retina pre-administered with DNL1.0, JIB-04, shikonin or avastin. The results are presented as means±S.E.M. (n = 4~ 10). The results of the ELISA assay are presented in picture b. The concentration of PLGF was measured in the retinas obtained from various groups, namely Sham, Vehicle+I/R, DNL1.0 + I/R or Eylea+I/R. Rats that received I/R plus pre-ischemia administration of 200 μg/5 μl Eylea was defined as the “Eylea+I/R” group. ** indicates a significant difference (P < 0.01) between the Sham retina and Vehicle+I/R. † or †† indicated a significant (P < 0.05 or P < 0.01) difference between Vehicle+I/R and DNL1.0 + I/R or between Vehicle+I/R and Eylea+I/R. Results are presented as means±S.E.M. (n = 4). Abbreviations: DNL, Dendrobium nobile Lindley. HIF-1α, hypoxia inducible factor-1α; VEGF, vascular endothelium factor; PKM2: pyruvate kinase M2, RBP2: retinoblastoma-binding protein 2; PLGF, placental growth factor