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Procyanidins on cerebral ischemia reperfusion injury of anti-inflammatory mechanisms

Abstract Objective To study the procyanidins (GSP) on focal cerebral ischemia and inflammatory mechanisms of reperfusion injury. Methods Rat model of cerebral ischemia after ischemic reperfusion 2 h 24 h, in which a large dose of proanthocyanidins (40 mg / kg), low dose group (10 mg / kg) 7 d before surgery by intraperitoneal injection 2 ml / kg GSP, model group, the intraperitoneal injection of 2 ml / kg saline, 1 / d, 15 min before reperfusion filling 1. Effect of different doses of proanthocyanidins on Brain Nitric Oxide (NO) levels, myeloperoxidase (MPO) activity and brain cell intercellular adhesion molecule -1 (ICAM-1) protein expression. Results procyanidins can significantly reduce the ischemia-reperfusion injury behavior score; significantly reduce NO content in brain tissue and MPO activity was significantly (P Key words procyanidins; cerebral ischemia-reperfusion injury; expression of intercellular adhesion molecule -1; myeloperoxidase; nitric oxide � 【Abstract】 Objective To investigate the Anti- Inflammatory mechanisms of grape seed procyanidin (GSP) on cerebral ischemia reperfusion injury (CIRI) of rats.Methods The model of MCAO was established by intraluminal suture technique.After 2 h of occlusion and 24 h of reperfusion, The high dose GSP (40 mg / kg) group and low dose GSP (10 mg / kg) group.was injected by intraperitoneal injection for 7 days, and all groups were given a more time in 15 minutes before operation.The contents of NO and activities of MPO in brain tissue of ischemic hemisphere were measured; the expression of ICAM-1 in the cortex were measured by immunohistochemical method.Results The neurological scores of rats treated with GSP was reduced apparently compared with model group rats (P <0.05); there were apparent reduction of the contents of NO and marked decrease of MPO in GSP group compared with the I / R model group (P <0.05); compared with I / R model group, the expression of ICAM-1 was down-regulated obviously in GSP group (P <0.05). Conclusion GSP could relieve the cerebral ischemia reperfusion injury, one of the mechanisms could be the Anti-Inflammatory effect of procyanidin � 【Key words】 Procyanidin; CIBI; ICAM-1; NO; MPO � is a kind of procyanidins from grape seed extract of natural plant polyphenols have strong antioxidant, scavenging free radicals, anti-inflammatory, reducing ischemia reperfusion injury [1 ] reduced platelet aggregation [2] such protection. Purpose of this study were to investigate the GSP in vivo cerebral ischemia and reperfusion damage (CIRI) and anti-inflammatory protective effect of injury mechanism of brain protection for clinical drug research and development to provide experimental evidence. � 1 � 1.1 Materials and methods Materials healthy Wistar rats were 64 and body mass 250 ~ 300 g, male, from Shandong Lukang Group Animal Center. Proanthocyanidins: Natural Product Research, Tianjin peak (batch number: G050804) to provide, ICAM-1 Immunohistochemistry Kit from Wuhan Boster Biological Engineering Co., Ltd. to provide; Coomassie brilliant blue protein assay kit, nitric oxide (NO) reagent box, myeloperoxidase (MPO) kits were purchased from Nanjing Jiancheng Biological Engineering Institute. Heraeus Instruments PRIM-R desktop high-speed cold centrifuge; Japan OLYMPUS BH-2 based biological company optical microscope and camera system; the Japanese company Sanyo MDF-330 Vertical ultra-low temperature refrigerators. � 1.2 model law prepared by modified Koizumi middle cerebral artery occlusion (MCAO) model. Preoperative fasting 12 h, free water, heating pads and surgical procedures using bulb to maintain body temperature in rats. Operation completed in 30 min, 2 h after ischemia, the filament out of reperfusion 24 h, recovery MCA blood supply. � 1.3 Reference Zea Longa neurological score of 5 points scoring system. � 1.4 Determination of sample preparation and target decapitated ischemic penumbra cortex adjacent to reserve. Electronic balance in brain tissue approximately 0.1 g, ice-cold saline flush residual blood, filter paper wipe adding Tris buffer with a glass homogenizer made of 10% brain tissue homogenate centrifuged at 4 ℃ 3500 turn / min ,[link widoczny dla zalogowanych], 15 min, supernatant NO,[link widoczny dla zalogowanych], MPO determination. Take the side penumbra of ischemic cortex fixed in fixative for immunohistochemistry samples produced. � 1.5 statistical methods were used for all data mean ± standard deviation (x ± s) that the application of SPSS 11.5 software was used to compare multiple samples analysis of variance,[link widoczny dla zalogowanych], P <0.05 as significant test. � 2 results � 2.1 GSP in rats CIRI GSP nerve function in large and small doses can improve the symptoms of neurological dysfunction, muscular strength, enhance, their behavior score and the model group The differences were statistically significant compared (P <0.05). � 2.2 GSP CIRI rat brain NO, MPO content compared with the sham group, model group, the NO content in brain tissue MPO activity was significantly increased, GSP large and small doses can significantly reduce the brain lacking blood reperfusion injury in rat brain serum NO, MPO content, and the model group,[link widoczny dla zalogowanych], the difference was significant (P <0.05), results in Table 1. � 2.3 GSP CIRI rats after brain ischemia Penumbra expression of ICAM-1 compared with model group, GSP dose group the expression of ICAM-1 expression was significantly decreased, showing small cytoplasmic There are brown granules, positive expression was significantly lower than the model group, low dose group and high dose group significantly reduced the number of positive cells,[link widoczny dla zalogowanych], the differences were statistically significant (P <0.05). The results in table 2, figure 1,2.
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