| Objective: We build the middle cerebral artery occlusion(MCAO)animal model to analysis the correlation between resveratrol dose and effect on animal brain model after injection.Method:1.Animal groups: to randomly divide 75 Sprague-Dawley(SD)rats,male,clean level,weight in the range of 220 ~ 250 g into the following 5 groups(15 each group): the model group,control group,low dose group(10 mg/kg),middle dose(30mg/kg)group,high dose group(60 mg/kg).2.To build the cerebral ischemia / reperfusion animal model: use chloral hydrate(10%)to anesthetize animal and stay in supine position,fix the animal on the table and give middle incision on animal neck area,to divide the external carotid artery and internal carotid artery,reveal the vagus nerve with glass needle.To insert the MCAO plug line after the animal neck external artery being incised,insert into neck artery around the common carotid artery bifurcation and slightly plug into arteries until slight resistance being felt(about 20 mm away from the bifurcation),to block the middle cerebral artery blood supply.After the ischemia time of model reach 90 min,pull out the MCAO line and restore the blood supply,and suture the wound after the reperfusion treatment and keep the animal into cages.Keep the rectum temperature of the animal stay at 37 ±0.5°C with heating pads.With following methods to build animal and could obtain rat neurons damaged model with higher stability,the results suggest repeatability.3.To detect the infarction area with TTC staining method To inject resveratrol with low dose(10 mg/kg),middle dose(30 mg/kg),high dose(60 mg/kg)through intraperitoneal injection 60 minutes after reperfusion.To immediately beheaded and extract the brain tissue after animal anaesthetized 24 hours after reperfusion,to slice 5 pieces(2 mm/piece)and keep them into TTC dye solution(2%),to process the staining under 37 ° C temperature for 30 minutes,to detect the brain infarction area.4.To detect the water content of brain area beheaded immediately and extract the brain tissue of ischemic brain one day after reperfusion,measure the weight and place into the oven continuously for 24 h at constant temperature(100 ° C),to measure the dry weight after thetissue dried.The water content in tissue could be calculated with following dry-wet method:(wet weight-dry weight)/wet weight by 100%.5.To observe the morphological characteristics of hippocampal neurons 7 days after reperfusion treatment.Results:Compared with the control group,ischemia reperfusion group often lead the model animals with higher brain water content;when compared with model group,different dose groups of animals show lower levels of brain tissue water content.The results of morphological characteristics were observed with light microscope on hippocampal neurons show that the control group could demonstrate full form of pyramidal neurons;anomaly characteristics such as degeneration,inflammation and necrosis can be observed in model group,resveratrol with middle dose injection could significant release the neuronal damage;resveratrol and ischemia would not influence the morphological characteristics of dentate gyrus.Conclusion: Resveratrol could effectively protect and improve the focal ischemia/reperfusion damage,the middle dose group could generate the best treat effect. |
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