Zhang Tong, Zhang Chunmei, Luo Yuyou, Liu Shuping, Li Siyuan, Li Lixia, Ma Yuxin, Liu Jing
Department of Basic Medicine, School of life sciences and biopharmaceutics, Guangdong Pharmaceutical University, Guangzhou 510006, China.
Department of Basic Medicine, School of life sciences and biopharmaceutics, Guangdong Pharmaceutical University, Guangzhou 510006, China.
Neurotoxicology. 2023 Mar;95:127-135. doi: 10.1016/j.neuro.2023.01.009. Epub 2023 Jan 16.
The present study aimed to investigate the protective effect of rutin on the injury of spinal motor neuron in rats exposed to acrylamide (ACR) the underlying mechanism. Fifty male Sprague-Dawley rats, aged 7-8 weeks, were randomly divided into control group, ACR group (20 mg/kg), low dose(100 mg/kg), medium dose (200 mg/kg) and high dose(400 mg/kg) rutin groups, ten rats in each group. The rats were given intragastric administration for 21 days. Every week, a neurobehavioral test was conducted. Nissl staining was used to observe the morphological changes in motor neurons in the L4-L6 segment of the spinal cord. Immunohistochemistry was used to identify AChE and ChAT in the rat spinal cord. Western blot was used to identify the expression of AChE, ChAT, P-ERK, ERK, and Nrf2 proteins in the rat spinal cord. The commercial kits were used to detect the presence of SOD, GSH, and LDH in the rat spinal cord. At the start of the second week, the medium and high dosage rutin group's rats' gait scores significantly decreased as compared to those of the ACR group. When rutin dosage was increased, the Nissl staining revealed that Nissl bodies was staining intensified compared to the ACR group. Immunohistochemistry and Western blot analysis revealed that AChE and ChAT expression changed when rutin dose was raised, but P-ERK and Nrf2 expression steadily increased in the spinal cord of rats in the medium and high dose groups compared to the ACR group. In the spinal cord of rats in each dosage group compared to the ACR group, the findings of the oxidative stress indices demonstrated that the expression levels of SOD and GSH rose with the increase of rutin dose, while the expression of LDH reduced with the rise of rutin dose. Rutin has an anti-oxidative impact through up-regulating the expression of P-ERK and Nrf2 proteins in the ERK/Nrf2 pathway, which may be connected to its protective action on motor neurons in the spinal cord of rats exposed to ACR.
本研究旨在探讨芦丁对丙烯酰胺（ACR）暴露大鼠脊髓运动神经元损伤的保护作用及其潜在机制。将50只7-8周龄的雄性Sprague-Dawley大鼠随机分为对照组、ACR组（20mg/kg）、低剂量（100mg/kg）、中剂量（200mg/kg）和高剂量（400mg/kg）芦丁组，每组10只。大鼠进行灌胃给药21天。每周进行一次神经行为学测试。采用尼氏染色观察脊髓L4-L6节段运动神经元的形态变化。采用免疫组织化学法鉴定大鼠脊髓中的乙酰胆碱酯酶（AChE）和胆碱乙酰转移酶（ChAT）。采用蛋白质免疫印迹法鉴定大鼠脊髓中AChE、ChAT、磷酸化细胞外信号调节激酶（P-ERK）、细胞外信号调节激酶（ERK）和核因子E2相关因子2（Nrf2）蛋白的表达。使用商业试剂盒检测大鼠脊髓中超氧化物歧化酶（SOD）、谷胱甘肽（GSH）和乳酸脱氢酶（LDH）的含量。在第二周开始时，中、高剂量芦丁组大鼠的步态评分与ACR组相比显著降低。随着芦丁剂量的增加，尼氏染色显示与ACR组相比尼氏体染色增强。免疫组织化学和蛋白质免疫印迹分析显示，随着芦丁剂量的增加，AChE和ChAT的表达发生变化，但与ACR组相比，中、高剂量组大鼠脊髓中P-ERK和Nrf2的表达稳步增加。与ACR组相比，各剂量组大鼠脊髓氧化应激指标结果显示，SOD和GSH的表达水平随芦丁剂量的增加而升高，而LDH的表达随芦丁剂量的增加而降低。芦丁通过上调ERK/Nrf2途径中P-ERK和Nrf2蛋白的表达发挥抗氧化作用，这可能与其对ACR暴露大鼠脊髓运动神经元的保护作用有关。
