目的: 观察小檗碱对大鼠脑缺血再灌注中NIMA相关蛋白激酶（NIMA-related Kinase,NEK7）/NLR家族Pxrin域蛋白3（nucleotide-blinding and leucinr-rich repeat proteins 3,NLRP3）信号通路的影响,探讨小檗碱在脑缺血再灌注损伤中的神经保护作用及机制。方法: 线栓法建立大鼠脑缺血再灌注损伤模型,随机分为假手术组、MCAO模型组、低剂量小檗碱组（25 mg/kg）、中剂量小檗碱组（50 mg/kg）、高剂量小檗碱组（100 mg/kg）,通过氯化三苯基四氮唑（triphenyltetrazolium chloride,TTC）染色测定脑梗死面积,HE染色观察脑组织形态学改变,ELISA法测定血清IL-1β和IL-18,RT-qPCR和Western-blot检测脑组织NEK7、NLRP3、Caspase-1的mRNA和蛋白表达水平。结果: 小檗碱组脑梗死体积较模型组减少,且中、高剂量小檗碱组大鼠脑梗死体积明显小于MCAO组。HE染色显示小檗碱组脑细胞损伤较模型组减轻。与MCAO组比较,小檗碱组大鼠血清中IL-1β和IL-18表达水平显著下降,小檗碱组脑组织NEK7、NLRP3、Caspase-1的mRNA和蛋白表达与模型组相比差异明显。结论: 小檗碱能显著减少脑梗死体积,减轻炎症反应,改善CIRI后神经功能缺损症状,其机制可能与NEK7/NLRP3通路有关。
Abstract
Objective To observe the effect of berberine on NEK7/NLRP3 signaling pathway in cerebral ischemia reperfusion, and to explore the neuroprotective effect and mechanism of berberine in cerebral ischemia reperfusion injury. Methods A rat cerebral ischemia reperfusion injury model was established by the wire bolus method, and randomly divided into a sham operation group, an MCAO model group, a low-dose berberine group (25 mg/kg), a medium-dose berberine group (50 mg/kg), and a high-dose berberine group (100 mg/kg), and the area of cerebral infarction was determined by TTC staining, the morphological changes of brain tissue were observed by HE staining, and the neuroprotective effects and mechanisms of berberine in cerebral ischemia reperfusion injury were measured by ELISA. The morphology of brain tissues was observed by HE staining, alteration, serum IL-1β and IL-18 were measured by ELISA, and the mRNA and protein expression levels of NEK7, NLRP3, and Caspase-1 in brain tissues were detected by RT-qPCR and Western-blot. Results The volume of cerebral infarction was reduced in berberine group compared with the model group, and the volume of cerebral infarction was significantly smaller in the middle and high dose berberine group than that in the MCAO group. HE staining showed that the damage of cerebral cells in the berberine group was reduced compared with that in the model group. Compared with the MCAO group, the expression levels of IL-1β and IL-18 in the serum of rats in the berberine group were significantly decreased, and the mRNA and protein expression of NEK7, NLRP3, and Caspase-1 in the brain tissues of the berberine group differed significantly from that of the model group. Conclusion Berberine can significantly reduce the volume of cerebral infarction, attenuate the inflammatory response, and improve the symptoms of neurological deficits after CIRI, and its mechanism may be related to the NEK7/NLRP3 pathway.
关键词
小檗碱 /
脑缺血再灌注 /
NIMA相关蛋白激酶7 /
NLRP3炎性小体
Key words
berberine /
cerebral ischemia reperfusion /
NIMA-related kinase 7 /
NLRP3 inflammasome
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基金
湖南省卫生健康委科技计划项目“小檗碱调控miR-143稳定动脉粥样硬化易损斑块的机制研究”（B202203073354）;湖南省自然科学基金“基于NEK7/NLRP3通路探讨小檗碱调节肠道菌群改善脑小血管病认知障碍的临床及基础研究”（2024JJ9304）
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