目的 探究丙酮酸乙酯（EP）调控硫氧还蛋白结合蛋白（TXNIP）/核苷酸结合域样受体蛋白3（NLRP3）/半胱氨酸天冬氨酸蛋白酶-1（Caspase-1）通路对缺氧缺血性脑损伤（HIBI）新生大鼠海马神经元焦亡的影响。方法 将SD大鼠随机分为假手术组，模型组，尼莫地平（8 mg·kg^-1）组，EP低、高剂量（1.5、3.0 mg·kg^-1）组，EP （3 mg·kg^-1）+白藜芦醇（Res，30 mg·kg^-1，TXNIP抑制剂）组。通过左颈总动脉结扎及缺氧处理构建HIBI大鼠模型，于造模24h后开始ip给药，每天1次，连续2周。采用Longa评分对各组大鼠神经功能损伤进行评估；ELISA法检测大鼠海马组织白细胞介素-1β（IL-1β）、白细胞介素-18（IL-18）含量；透射电子显微镜下观察大鼠海马超微结构；HE染色观察海马组织病理学情况；TUNEL染色观察海马神经元凋亡；Western blotting检测海马组织中TXNIP/NLRP3/Caspase-1通路相关蛋白表达。结果 与假手术组比较，模型组Longa评分、海马组织IL-1β和IL-18水平、神经元凋亡指数（AI）、以及TXNIP、NLRP3、凋亡相关斑点样蛋白（ASC）、cleaved Caspase-1蛋白表达水平显著增加（P<0.05），神经元损伤、海马组织病理学程度明显增加；与模型组比较，尼莫地平组和EP低、高剂量组Longa评分、IL-1β和IL-18水平、神经元AI以及TXNIP、NLRP3、ASC、cleaved Caspase-1蛋白表达水平显著降低（P<0.05），神经元损伤、海马组织病理学程度明显改善；与EP高剂量组比较，EP+Res组Longa评分、IL-1β和IL-18水平、神经元AI以及TXNIP、NLRP3、ASC、cleaved Caspase-1蛋白表达水平显著降低（P<0.05），神经元损伤、海马组织病理学程度改善更明显。结论 EP可能通过抑制TXNIP/NLRP3/Caspase-1通路来减轻HIBI新生大鼠海马神经元焦亡。

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

河北省科技计划项目（17277714D）