Postischemic administration of liposome-encapsulated luteolin prevents against ischemia-reperfusion injury in a rat middle cerebral artery occlusion model

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Postischemic administration of liposome-encapsulated luteolin prevents against ischemia-reperfusion injury in a rat middle cerebral artery occlusion model Gang Zhao, Shao-Yun Zang, Zhi-Hua Jiang, Yao-Yue Chen, Xun-He Ji, Bu-Feng Lu, Jia-Hu Wu, Guo-Wei Qin, Li-He Guo Journal of Nutritional Biochemistry Volume 22, Issue 10, Pages 929-936 (October 2011) DOI: 10.1016/j.jnutbio.2010.07.014 Copyright © 2011 Elsevier Inc. Terms and Conditions

Fig. 1 Schematic drawing of the thread placed into the ECA with its tip proximal to the origin of the MCA. CCA, common carotid artery; PCA, posterior cerebral artery; VA, vertebral artery; BA, basilar artery; OpA, ophthalmic artery. Journal of Nutritional Biochemistry 2011 22, 929-936DOI: (10.1016/j.jnutbio.2010.07.014) Copyright © 2011 Elsevier Inc. Terms and Conditions

Fig. 2 Neuroprotective actions of luteolin (LU) and vitamin E (VE) on H2O2-insulted neonatal neurons (by CCK-8 test). HP, hydrogen peroxide (H2O2). Activity of each cell group was represented as the OD ratio between treatment cell group and concentration-paired, no-cell group. One-way ANOVA, F7,16=275.45, P<.001. Post-hoc, ⁎⁎P<.01 vs. HP-only; @P<.05 vs. HP plus VE 15 μM. Values are expressed as mean±S.E.M. of triplicate cell samples. Journal of Nutritional Biochemistry 2011 22, 929-936DOI: (10.1016/j.jnutbio.2010.07.014) Copyright © 2011 Elsevier Inc. Terms and Conditions

Fig. 3 Effect of luteolin on symptom score of I/R rats. Symptom score of each animal was rated at 10 h (4 h after treatment onset), on Day 7 and on Day 14 after reperfusion. At each time point, F3,64=28.45, 55.3 and 147.36, respectively for the time points of 10 h, Day 7 and Day 14, all P<.01 (by one-way ANOVA, same as mentioned below). Post hoc, ⁎P<.05 and ⁎⁎P<.01 and, respectively, vs. corresponding day-paired model group (by LSD, same as mentioned below). Data were expressed as mean±S.E.M. of 16–18 animals. Journal of Nutritional Biochemistry 2011 22, 929-936DOI: (10.1016/j.jnutbio.2010.07.014) Copyright © 2011 Elsevier Inc. Terms and Conditions

Fig. 4 Effect of luteolin on the balance performance of I/R rats. Balance behavior of each animal was evaluated at 10 h, on Day 7 and on Day 14 after reperfusion. At each time point, F3,64=47.11, 103.51 and 237.06, respectively, for the 1, 7 and 14-day time point, all P<.01. Post hoc, ⁎P<.05 and ⁎⁎P<.01, respectively, vs. same-day-paired model group. Data were expressed as mean±S.E.M. of 16–18 animals. Journal of Nutritional Biochemistry 2011 22, 929-936DOI: (10.1016/j.jnutbio.2010.07.014) Copyright © 2011 Elsevier Inc. Terms and Conditions

Fig. 5 Effect of luteolin on TTC stain (A) and the infarction valume (B) of I/R rats. TTC staining was performed after the final behavioral test, and infarction volume of each slab was calculated. Data were expressed as percent of volume of contralateral hemisphere (mean±S.E.M. of 8–9 animals). F3,30=43.75 and F3,30=59.16, respectively, for hemisphere and striatum, both P<.01. Post hoc, ⁎P<.05 and ⁎⁎P<.01, respectively, vs. respective model group. Journal of Nutritional Biochemistry 2011 22, 929-936DOI: (10.1016/j.jnutbio.2010.07.014) Copyright © 2011 Elsevier Inc. Terms and Conditions

Fig. 6 Protective action of luteolin on neuronal cells in hippocampus and ischemic penumbra in I/R rats (assayed by cresyl violet staining). (A) Sections of hippocampus with scale bar of 200 μm; arrows, indicating the regions enlarged for Fig. 6B. (B) Sections of hippocampus with scale bar=20 μm. (C) Cell shape in ischemic penumbral region of parietal cortex (scale bar=20 μm); parietal cortex in sham, serving as control. Panels I, II, III and IV in column A, B or C, representing sham, model, LipLU 5 mg/kg, and LipLU 20 mg/kg group, respectively. DG, dentate gyrus; Δ, injured regions. Journal of Nutritional Biochemistry 2011 22, 929-936DOI: (10.1016/j.jnutbio.2010.07.014) Copyright © 2011 Elsevier Inc. Terms and Conditions

Fig. 7 Effect of luteolin on the level of ROS (DCF) in I/R rats' brain. DCF levels were assayed upon the mitochondria prepared from the ipsilateral hippocampus, frontal cortex and striatum of rats. ⁎P<.05 and ⁎⁎P<.01, respectively, vs. respective model group. Data were expressed as mean±S.E.M. of eight to nine animals. Journal of Nutritional Biochemistry 2011 22, 929-936DOI: (10.1016/j.jnutbio.2010.07.014) Copyright © 2011 Elsevier Inc. Terms and Conditions

Fig. 8 Effect of luteolin on the level of CAT and reduced GSH of I/R rat brain. Levels of CAT (A) and GSH (B) in ipsilateral hippocampus, frontal cortex and striatum were assayed by using the CAT and GSH kits. ⁎P<.05 and ⁎⁎P<.01, respectively, vs. respective model group. Data were expressed as mean±S.E.M. of eight to nine animals. Journal of Nutritional Biochemistry 2011 22, 929-936DOI: (10.1016/j.jnutbio.2010.07.014) Copyright © 2011 Elsevier Inc. Terms and Conditions