目的 探究丙酮酸乙酯（EP）調(diào)控硫氧還蛋白結(jié)合蛋白（TXNIP）/核苷酸結(jié)合域樣受體蛋白3（NLRP3）/半胱氨酸天冬氨酸蛋白酶-1（Caspase-1）通路對(duì)缺氧缺血性腦損傷（HIBI）新生大鼠海馬神經(jīng)元焦亡的影響。方法 將SD大鼠隨機(jī)分為假手術(shù)組，模型組，尼莫地平（8 mg·kg^-1）組，EP低、高劑量（1.5、3.0 mg·kg^-1）組，EP （3 mg·kg^-1）+白藜蘆醇（Res，30 mg·kg^-1，TXNIP抑制劑）組。通過左頸總動(dòng)脈結(jié)扎及缺氧處理構(gòu)建HIBI大鼠模型，于造模24h后開始ip給藥，每天1次，連續(xù)2周。采用Longa評(píng)分對(duì)各組大鼠神經(jīng)功能損傷進(jìn)行評(píng)估；ELISA法檢測(cè)大鼠海馬組織白細(xì)胞介素-1β（IL-1β）、白細(xì)胞介素-18（IL-18）含量；透射電子顯微鏡下觀察大鼠海馬超微結(jié)構(gòu)；HE染色觀察海馬組織病理學(xué)情況；TUNEL染色觀察海馬神經(jīng)元凋亡；Western blotting檢測(cè)海馬組織中TXNIP/NLRP3/Caspase-1通路相關(guān)蛋白表達(dá)。結(jié)果 與假手術(shù)組比較，模型組Longa評(píng)分、海馬組織IL-1β和IL-18水平、神經(jīng)元凋亡指數(shù)（AI）、以及TXNIP、NLRP3、凋亡相關(guān)斑點(diǎn)樣蛋白（ASC）、cleaved Caspase-1蛋白表達(dá)水平顯著增加（P<0.05），神經(jīng)元損傷、海馬組織病理學(xué)程度明顯增加；與模型組比較，尼莫地平組和EP低、高劑量組Longa評(píng)分、IL-1β和IL-18水平、神經(jīng)元AI以及TXNIP、NLRP3、ASC、cleaved Caspase-1蛋白表達(dá)水平顯著降低（P<0.05），神經(jīng)元損傷、海馬組織病理學(xué)程度明顯改善；與EP高劑量組比較，EP+Res組Longa評(píng)分、IL-1β和IL-18水平、神經(jīng)元AI以及TXNIP、NLRP3、ASC、cleaved Caspase-1蛋白表達(dá)水平顯著降低（P<0.05），神經(jīng)元損傷、海馬組織病理學(xué)程度改善更明顯。結(jié)論 EP可能通過抑制TXNIP/NLRP3/Caspase-1通路來減輕HIBI新生大鼠海馬神經(jīng)元焦亡。

Objective To explore the effect of ethyl pyruvate(EP) on hippocampal neuronal pyrolysis in neonatal hypoxic-ischemic rats by regulating the thioredoxin-interacting protein(TXNIP)/nucleotide-binding domain-like receptor 3(NLRP3)/cysteine aspartate proteinase-1(Caspase-1) pathway.Methods SD rats were randomly divided into sham-operation group, model group, nimodipine(8 mg·kg^-1), EP-L(1.5 mg·kg^-1) group, EP-H(3 mg·kg^-1) group, EP-H(3 mg·kg^-1)+Res(30 mg·kg^-1, TXNIP inhibitor Res) group, with 15 rats in each group.HIBI rat model was constructed by ligation of the left common carotid artery and hypoxia treatment.Rats were ip with corresponding doses once a day for two consecutive weeks after 24 hours of modeling.Longa score was used to evaluate the neurological damage of rats in each group.ELISA was used to detect the levels of interleukin 1β(IL-1β) and interleukin18(IL-18) in the hippocampus of rats.A transmission electron microscope was used to observe the ultrastructure of rat hippocampus.HE staining was used to observe the pathology of hippocampus.TUNEL staining was used to observe the apoptosis of hippocampal neurons.Western blotting was used to detect the expression of TXNIP/NLRP3/Caspase-1 pathway related proteins in hippocampus.Results Compared with the sham operation group, the nerve function damage scores, hippocampal IL-1β and IL-18 levels, neuron damage, hippocampal histopathology, neuronal apoptosis index(AI), TXNIP, NLRP3, apoptosis-related speck-like protein(ASC) and cleaved Caspase-1 protein expression levels increased significantly in model group(P<0.05).Compared model group, the nerve function damage scores, IL-1β and IL-18 levels, neuron damage, hippocampal histopathology, neuronal AI, TXNIP, NLRP3, ASC and cleaved Caspase-1 protein expression levels decreased significantly in the nimodipine group, EP-L group, EP-H group(P<0.05).Compared with EP-H group, the nerve function damage scores, IL-1β and IL-18 levels, neuron damage, hippocampal histopathology, neuronal AI, TXNIP, NLRP3, ASC and cleaved Caspase-1 protein expression levels decreased significantly in the EP-H+Res group(P<0.05).Conclusion EP may inhibit the TXNIP/NLRP3/Caspase-1 pathway to reduce the pyroly sis of hippocampal neurons in neonatal rats with HIBI.

R965

河北省科技計(jì)劃項(xiàng)目（17277714D）