Protective Effects And The Mechanisms Of D-allose Pretreatment Against Cerebral Ischemia Injury In Mice

Ischemic stroke, a complex devastating disease, is caused by a drastic disruption of cerebral blood flow that triggers amultistep pathophysiological ischemic sufficient to cause metabolic disorder or functional deficit. cerebral is chemia-reperfusion injury causes great harm. For an ischemic-reperfusion injury, emergency treatment focuses on therapeutic method to restore blood flow, but it fails to reverse brain damage even if it restores blood flow. Cerebral ischemia causes neuron necrosis, which induces an inflammatory response, moreover, cerebral ischemia-reperfusion produces a large amount of active oxygen free radicals, what is more, the inflammatory cell s in ischemia area lead to an increased production of Inflammatory cytokines, chemokines, and adhesion molecules, furthermore leading to leucocytes trapping in capillary, which would induce low flow perfusion. Our findings suggest that D-allose carries a therapeutic potential in ischemic stroke possibly.D-allose, a type of rare sugar, exists rarely in nature but has been isolated from the leaves of the African shrub Protea rubropilosa. D-Allose has attracted a great deal of attention in recent years due to its many biochemical properties, which include s hypolipidemic, hypoglycemia, Free radical scavenging activity, neuroprotective effect and anti-tumor effect. Now research has shown that D-allose has an immunoprotective effect on the liver and kidney, furthermore, has positive effect against ischemia-reperfusion injury on the rat abdominal skin island flap model, So the D-allose research has become a hot spot in focus in recent years. In order to understand protective effect of D-allose against cerebral ischemia reperfusion injury in mice, We make a further research to delve deeper into their conversations.This experimental study includes two parts. part one, experiment studys the effect of D-allose on the brain in the mice mode of MCAO. Part two, it explores its possible mechanisms about protective action of D-allose against cerebral ischemia reperfusion injury in mice.Part 1 protective effects of D-allose pretreatment against cerebral ischemia reperfusion injury in miceObjective To study the effect of D-allose against cerebral ischemia reperfusion injury in mice.Metheds The focal cerebral ischemia-reperfusion model was induced by the middle cerebral artery occlusion(MCAO). Sixty male Balb/c mice were randomly divided into sham group(fifteen mice), MCAO group(fifteen mice), MCAO+NS group(fifteen mice), MCAO+D-allose group(fifteen mice). With a model of reversible middle cerebral artery occlusion in mice, D-allose was injected in caudal vein(0.4 mg/g) before its surgery. the operating time was 20 minute,the time of MCAO was 120 min. The neurological deficit score was performed by modified neurolo gical severity scores(m NSS), the infarct volume was calculated by triphenyl tetrazolium chloride(TTC) Stain. The brain water content was measured by Dry/Wet method. Blood-brain barrier(BBB) permeability was evaluated by Evans blue(EB) extravasation. The histopathological changes was observed by HE staining and Nissl staining, the expression of matrix metalloproteinase-9(MMP-9) was detected by immunohistochemistry method.Results Compared with MCAO group, D-allose group significantly lowered neurological deficit score(P<0.05), in addition, could reduce the infarct volume(P<0.05) and alleviate brain water content markedly(P<0.05), what is more, D-allose diminished the extent and quantity of Evans blue extravasation(P<0.05). Eventually, Compared with MCAO group, D-allose decreased the level of MMP-9 expression(P<0.05) and ameliorated the histopathological changes(P<0.05).Conclusion D-allose has the protective effects on the ischemic brain area following focal cerebral ischemia reperfusion, which is closely related to inhibition of MMP-9 expression, furthermore, it protects the blood brain barrier. Part 2 the mechanisms relative to the protective effects of D-allose pretreatment against Cerebral Ischemia Injury in miceObjective To study the mechanisms relative to the protective effects of D-allose pretreatment against Cerebral Ischemia Injury in mice.Metheds Thirty-five male Balb/c mice were randomly divided into sham group(ten mice), MCAO group(ten mice), MCAO+D-allose group(ten mice), MCAO+GW9662 group(five mice). With a model of reversible middle cerebral artery occlusion in mice, TNF-?,IL-1? was detect by ELISA. the positive cells of TNF-?, NF-k B, ICAM-1 and VCAM-1was assessed by immunohistochemistry method. The protein expression of PPAR?, NF-k B, TNF-? was tested by western blot.Results immunohistochemistry method showed that D-allose could reduce the positive cells number of TNF-?, NF-k B, ICAM-1 and VCAM-1(P<0.05), in addition, ELISA tests showed that D-allose group significantly lowered the protein expression TNF-?,IL-1?(P<0.05), The western blot showed that D-allose could improve the protein expression of PPAR?,but contrary to reduce the protein expression of NF-k B and TNF-?(P<0.05). in addition, MCAO+GW9662 group colud Lower the protein expression of PPAR?, Instead, it colud aggrandize the protein expression of NF-k B and TNF-?.Conclusion D-allose exerts neuroprotective effects against focal cerebral ischemia/ reperfusion in mice because of its anti-inflammatory mechanism.