基于AMPK/eNOS/NF-κB信号通路探讨虾青素对脑梗死大鼠的影响及作用机制  

Effect of astaxanthin on cerebral infarction rats and its mechanism based on AMPK/eNOS/NF-κB signal pathway

作　　者：吴杰[1] 杜彩萍 WU Jie;DU Caiping(Department of Pharmaceutical Technology,Jiangsu Provincial Xuzhou Pharmaceutical Vocational College,Xuzhou,Jiangsu 221116,China;2.Research Center for Biochemistry and Molecular Biology,School of Basic Medicine,Xuzhou Medical University,Xuzhou,Jiangsu 221004,China)

机构地区：[1]江苏省徐州医药高等职业学校药学技术系,江苏省徐州市221116 [2]徐州医科大学基础医学院生物化学与分子生物学研究中心,江苏省徐州市221004

出　　处：《中国动脉硬化杂志》2018年第12期1221-1226,共6页Chinese Journal of Arteriosclerosis

基　　金：国家自然科学基金项目(81100852)

摘　　要：目的基于AMPK/eNOS/NF-κB信号通路探讨虾青素对脑梗死大鼠的影响及作用机制。方法采用改良的Longa法建立脑梗死大鼠模型,取50只造模成功的SD雄性大鼠随机分为模型对照组、阳性对照组、虾青素低、中、高剂量组,每组10只,另外未造模的10只作为空白对照组。空白对照组与模型对照组均给予等体积生理盐水,阳性对照组给予20mg/(kg·d)尼莫地平,虾青素低、中、高剂量组分别给予10、20、30mg/(kg·d)虾青素,连续给药14天。检测大鼠神经功能变化;氯化三苯基四氮唑染色法检测脑梗死面积;酶联免疫吸附法检测白细胞介素10(IL-10)、IL-1β、IL-6和肿瘤坏死因子α(TNF-α)水平;TUNEL法检测脑皮质缺血半影区的细胞凋亡情况;反转录-聚合酶链反应检测腺苷酸活化蛋白激酶(AMPK)、内皮型一氧化氮合酶(eNOS)、核因子κB(NF-κB)mRNA表达水平;Westernblot检测p-AMPK、AMPK、p-eNOS、eNOS、NF-κB蛋白表达。结果虾青素可明显改善脑梗死大鼠的神经功能及脑梗死面积(P<0.01);抑制促炎因子IL-1β、IL-6和TNF-α表达,促进抗炎因子IL-10表达(P<0.05);降低皮质缺血半影区细胞凋亡率(P<0.01);下调NF-κB mRNA和蛋白表达,促进AMPK和eNOS磷酸化(P<0.05),但AMPK、eNOSmRNA和蛋白表达无显著差异(P>0.05)。结论虾青素能明显改善脑梗死大鼠神经功能、脑梗死面积、皮质缺血半影区细胞凋亡率及血清细胞因子,其作用机制可能与AMPK/eNOS/NF-κB信号通路密切相关。Aim To explore the effect and mechanism of astaxanthin on cerebral infarction rats based on AMPK/eNOS/NF-kappa B signal pathway. Methods The rat model of cerebral infarction was established by modified Longa method. 50 successful modeling SD male rats were randomly divided into model control group,positive control group, astaxanthin low,medium and high dose group,with 10 rats in each group,and 10 rats without model as blank control group. The blank control group and the model control group were given the same volume of normal saline,the positive control group was given 20 mg /( kg·d) nimodipine,and the astaxanthin low,medium and high dose groups were given 10,20,30 mg /( kg·d) astaxanthin,respectively,for 14 consecutive days. Neurological changes were detected in rats; cerebral infarction area was measured by triphenyl tetrazolium chloride staining; levels of interleukin-10 ( IL-10) ,interleukin- 1β,interleukin-6 and tumor necrosis factor-α ( TNF-α) were detected by enzyme-linked immunosorbent assay; cell apoptosis in ischemic penumbra of cerebral cortex was detected by TUNEL; the expressions of adenosine monophosphate activated protein kinase ( AMPK) ,endothelial nitric oxide synthase ( eNOS) and nuclear factor κB ( NF-κB) mRNA were measured by reverse transcription-polymerase chain reaction ( RT-PCR) ; the protein expressions of p-AMPK,AMPK,peNOS, eNOS and NF-κB were detected by Western blot. Results Astaxanthin could significantly improve neurological function and cerebral infarction area in rats with cerebral infarction ( P<0.01) ,inhibit the expression of pro-inflammatory factors IL-1β,IL-6 and TNF-α,promote the expression of anti-inflammatory factor IL-10 ( P<0.05) ,reduce the cell apoptosis rate of cortical ischemic penumbra ( P<0.01) ,down-regulate the expressions of NF-κB mRNA and protein,and promote phosphorylation of AMPK and eNOS ( P<0.05) ,but there was no significant difference in the expressions of AMPK, eNOS mRNA and protein ( P > 0. 05) . Conclusion Astaxanthin can significantly improv

关 键 词：AMPK/eNOS/NF-κB信号通路 虾青素 脑梗死 神经再生 

分 类 号：R96[医药卫生—药理学]