目的 探究利多卡因?qū)δ摱景Y相關(guān)性腦病模型大鼠的腦保護(hù)作用及機(jī)制。方法 將40只大鼠按隨機(jī)數(shù)字表法分為對照組、模型組、利多卡因組、利多卡因＋血管緊張素受體AT1相關(guān)的受體蛋白拮抗劑Apelin13（F13A）組，每組10只。除對照組外，其余3組大鼠均構(gòu)建膿毒癥相關(guān)性腦病模型，利多卡因組和利多卡因＋F13A組造模后即刻給予利多卡因10 mg/kg負(fù)荷劑量，隨后尾iv利多卡因10 mg/kg并持續(xù)3 h，利多卡因＋F13A組同時(shí)ip APJ拮抗劑F13A 100 μg/kg。24 h后，Morris水迷宮實(shí)驗(yàn)檢測各組大鼠認(rèn)知功能，酶聯(lián)免疫吸附（ELISA）法檢測各組大鼠血清白細(xì)胞介素（IL）-6、IL-10、腫瘤壞死因子-α（TNF-α）水平，蘇木精-伊紅（HE）染色觀察各組大鼠腦組織病理變化，TdT介導(dǎo)的dUTP缺口末端標(biāo)記法（TUNEL）和神經(jīng)元特異核蛋白（NeuN）雙免疫熒光染色檢測各組大鼠腦組織皮質(zhì)內(nèi)神經(jīng)元凋亡，免疫熒光雙染法觀察各組大鼠腦組織皮質(zhì)內(nèi)Apelin與APJ表達(dá)，實(shí)時(shí)熒光定量逆轉(zhuǎn)錄聚合酶鏈反應(yīng)（RT-qRCR）檢測各組大鼠腦組織Apelin和APJ的mRNA表達(dá)，蛋白質(zhì)免疫印跡（Western blotting）法檢測各組大鼠腦組織Apelin和APJ的蛋白表達(dá)。結(jié)果 與模型組比較，利多卡因組大鼠逃避潛伏期縮短，通過目標(biāo)象限次數(shù)增加（P＜0.05），血清IL-6、TNF-α水平降低而IL-10水平顯著升高（P＜0.05），海馬區(qū)神經(jīng)元損傷明顯減輕，形態(tài)和分布均趨于正常，腦組織皮質(zhì)內(nèi)凋亡神經(jīng)元數(shù)目減少（P＜0.05）；Apelin與APJ熒光染色表達(dá)均明顯增強(qiáng)，腦組織內(nèi)Apelin、APJ的mRNA與蛋白相對表達(dá)量均顯著上調(diào)（P＜0.05）；與利多卡因組比較，利多卡因＋F13A組大鼠逃避潛伏期延長，通過目標(biāo)象限次數(shù)減少（P＜0.05），血清IL-6、TNF-α水平升高，血清IL-10水平顯著降低（P＜0.05），海馬區(qū)神經(jīng)元損傷加重，有大量細(xì)胞腫脹與細(xì)胞核固縮，腦組織皮質(zhì)內(nèi)凋亡神經(jīng)元數(shù)目增加（P＜0.05）。同時(shí)，Apelin與APJ熒光染色表達(dá)均明顯減弱，腦組織內(nèi)Apelin、APJ的mRNA與蛋白相對表達(dá)量均顯著下調(diào)（P＜0.05）。結(jié)論 利多卡因能夠改善膿毒癥相關(guān)性腦病模型大鼠腦損傷，抑制炎癥反應(yīng)，并減少神經(jīng)元凋亡，其機(jī)制可能與激活A(yù)pelin/APJ系統(tǒng)有關(guān)。

Objective To investigate the protective effects and mechanism of lidocaine on sepsis-associated encephalopathy model rats. Methods Forty rats were randomly divided into control group, model group, lidocaine group, and lidocaine + F13A group, with 10 rats in each group. Except the control group, the other three groups of rats were constructed sepsis-associated encephalopathy model, lidocaine group and lidocaine + F13A group were given a loading dose of 10 mg/kg lidocaine immediately after modeling, after that, lidocaine 10 mg/kg was injected intravenously for 3 h, APJ antagonist F13A 100 μg/kg was injected intraperitoneally in lidocaine + F13A group. 24 h later, the Morris water maze test detected the cognitive function of each group, enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of serum interleukin(IL)-6, IL-10, and tumor necrosis factor-α (TNF-α), hematoxylin-eosin (HE) staining was used to observe pathological changes in brain tissue of rats in each groups, TdT mediated dUTP nick end labeling (TUNEL) and neuron-specific nuclear protein (NeuN) double immunofluorescence staining was used to detect neuronal apoptosis in the cerebral cortex of each group, the expressions of Apelin and APJ in the cerebral cortex of rats in each group were observed by immunofluorescence double staining, real-time polymerase chain reaction (RT-qRCR) was used to detect the mRNA expressions of Apelin and APJ in the brain tissues of rats in each group, Western blotting was used to detect the protein expressions of Apelin and APJ in the brain tissues of rats in each group. Results Compared with model group, the escape latency of rats in lidocaine group was shortened, the number of target quadrant was increased (P < 0.05), the levels of IL-6 and TNF-α in serum were decreased while the level of IL-10 was increased (P < 0.05), the damage of neurons in hippocampus was significantly reduced, and the morphology and distribution of neurons tended to be normal, the number of apoptotic neurons in the cerebral cortex was decreased (P < 0.05), the expression of Apelin and APJ fluorescence staining were significantly increased, the mRNA and protein relative expression levels of Apelin and APJ in the cerebral tissue were also significantly up-regulated (P < 0.05). Compared with lidocaine group, the escape latency of rats in lidocaine + F13A group was prolonged, the number of passage through the target quadrant was reduced (P < 0.05), the levels of IL-6 and TNF-α in serum were increased, and the level of IL-10 in serum were decreased (P < 0.05), the neuronal damage in hippocampus was aggravated, there was a large number of cell swelling and nuclear pyretosis, the number of apoptotic neurons in the cerebral cortex was increased (P < 0.05). Meanwhile, the expression of Apelin and APJ fluorescence staining were significantly decreased, and the mRNA and protein relative expression levels of Apelin and APJ in the cerebral tissue were significantly decreased (P < 0.05). Conclusions lidocaine can improve brain injury, inhibit inflammation and reduce neuronal apoptosis in sepsis-associated encephalopathy model rats, and the mechanism may be related to the activation of Apelin/APJ system.

R965

新疆維吾爾自治區(qū)衛(wèi)生健康青年醫(yī)學(xué)科技人才專項(xiàng)科研項(xiàng)目（WJWY-202203）