目的 研究藏紅花素預(yù)處理對(duì)大鼠全腦缺血再灌注損傷的保護(hù)作用及其機(jī)制。方法 采用四動(dòng)脈阻斷法復(fù)制全腦缺血再灌注大鼠模型；藏紅花素低、中、高劑量組分別于術(shù)前7 d開(kāi)始1次/d ip給予10、20、40 mg/kg藏紅花素，另設(shè)假手術(shù)組、模型組和尼莫地平（陽(yáng)性藥，1 mg/kg）組。測(cè)定腦電圖恢復(fù)時(shí)間和翻正反射恢復(fù)時(shí)間；術(shù)后24 h評(píng)卒中指數(shù)和神經(jīng)功能缺失評(píng)分；干濕比重法測(cè)定腦組織含水量；伊文氏藍(lán)法檢測(cè)血腦屏障（BBB）通透性；HE染色法行大腦皮層神經(jīng)元病理學(xué)檢查并進(jìn)行病變分級(jí)；末端標(biāo)記法（TUNEL）觀察皮層神經(jīng)元凋亡狀況并計(jì)算凋亡指數(shù)；生化分析法測(cè)定氧自由基（ROS）、丙二醛（MDA）含量及超氧化物歧化酶（SOD）、過(guò)氧化氫酶（CAT）活性；酶聯(lián)免疫吸附法（ELISA）測(cè)定白介素（IL）-1β、腫瘤壞死因子（TNF-α）、IL-6、干擾素-γ（IFN-γ）水平。結(jié)果 與模型組比較，藏紅花素中、高劑量或尼莫地平預(yù)處理均明顯縮短全腦缺血性腦損傷大鼠腦電圖恢復(fù)時(shí)間和翻正反射恢復(fù)時(shí)間，降低卒中指數(shù)和神經(jīng)功能缺失評(píng)分，抑制腦組織含水量升高和伊文氏藍(lán)滲入，抑制缺血大腦皮層神經(jīng)元病理性形態(tài)結(jié)構(gòu)改變和病變等級(jí)的升高，抑制缺血大腦皮層神經(jīng)元凋亡、降低凋亡指數(shù)，抑制ROS、MDA含量升高和SOD、CAT活性降低，抑制TNF-α、IL-1β、IL-6、IFN-γ含量升高，差異均具有統(tǒng)計(jì)學(xué)意義（P<0.05、0.01）。結(jié)論 藏紅花素預(yù)處理對(duì)大鼠全腦缺血再灌注損傷具有保護(hù)作用，機(jī)制可能與抑制氧化應(yīng)激損傷和抑制炎癥級(jí)聯(lián)反應(yīng)有關(guān)。

Objective To study the protective effect and mechanism of crocin pretreatment on global cerebral ischemia reperfusion in rats. Methods Four-artery occlusion was used to duplicate the rat model of cerebral ischemic injury. 7 days before operation, the low, medium and high dose crocin pretreatment groups were given 10, 20 and 40 mg/kg by ip injection once a day; another sham operation group, model group and nimodipine pretreatment group (positive drug, 1 mg/kg) were set up. The electroencephalogram recovery time and the righting reflex recovery time were measured; 24 h after operation, the stroke score and neurological symptom score were performed, the water content in brain tissue were determined by dry-wet specific gravity method, the permeability of blood brain barrier(BBB) was detected by evans blue method, the pathological examination of cerebral cortical neurons by HE staining and the pathological grading was carried out, the apoptosis of cortical neurons was detected by TUNEL and the apoptosis indes was calculated; the content of oxygen free radicals (ROS), MDA and the activities of superoxide dismutase (SOD) and catalase (CAT) were determined by biochemical analysis; the inflammatory cytokines content of IL-1β, TNF-α, IL-6, IFN-γ were determined by ELISA. Results Compared with model control group, the pretreatment of crocin with medium, high dose or nimodipine could shorten the recovery time of EEG and righting reflex in rats with cerebral ischemic injury, reduce stroke score and neurological symptom score, inhibit the increase of brain tissue water content and the infiltration of evans blue, inhibit pathological morphological and structural changes of cerebral cortical neurons on ischemic side, inhibit the apoptosis of ischemic cerebral neuronal and reduce the apoptosis index, inhibit the increase of ROS, MDA content and the decrease of SOD, CAT activities, Inhibit the increase of TNF-α, IL-1β, IL-6, IFN-γ content; all of the difference was statistically significant (P < 0.05 or 0.01). Conclusion crocin pretreatment has a protective effect on global cerebral ischemic reperfusion injury, and the mechanism may be related to inhibition of oxidative stress injury and inhibition of inflammatory response.

R965

河北省中醫(yī)藥管理局科研計(jì)劃項(xiàng)目（2019367）