天麻素对糖氧剥夺再复供皮层神经细胞NF-κB炎症级联信号通路表达的影响  

Effect of Gastrodin on Expression of NF-κB Inflammatory Cascade Signaling Pathway in Cortical Neurons After Oxygen-glucose Deprivation and Reperfusion

作　　者：文欢[1] 张大燕[1] 王伟[1] 高继海[1] 饶朝龙[1] 彭成[1] 

机构地区：[1]成都中医药大学药学院,中药资源系统研究与开发利用省部共建国家重点实验室培育基地,成都611137

出　　处：《中国实验方剂学杂志》2017年第21期104-111,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金人才培养项目(J1310034);中药资源四川省青年科技创新研究团队项目(2015TD0028);四川省中医药管理局项目(2016Z008)

摘　　要：目的:研究天麻素对糖氧剥夺再复供损伤原代皮层神经细胞的保护作用,以及对炎症相关信号通路表达的影响。方法:构建皮层神经细胞糖氧剥夺再复供(OGD/R)损伤模型,随机分为空白组、模型组、天麻素(80,60,40 mg·L-1)组。噻唑蓝(MTT)比色法测定皮层细胞存活率,比色法测定乳酸脱氢酶(LDH)漏出率,高通量RNA-Seq测序分析技术检测处理细胞的所有基因转录本的表达差异,通过功能注释与富集分析筛选出与OGD/R损伤或天麻素预处理保护作用相关的基因分子与代谢通路。结果:OGD/R损伤后皮层细胞活性下降,LDH释放增加,而天麻素预处理能提高细胞存活率,并显著下调LDH漏出率,转录组测序分析发现其分子机制主要涉及核转录因子-κB(NF-κB,18 gene),肿瘤坏死因子(TNF,21 gene)等信号通路的抑制调控。与模型组比较,天麻素下调了NF-κB,Toll样受体2(TLR2),肿瘤坏死因子受体1/2(TNFR1/2),白细胞介素-6(IL-6),细胞间黏附分子-1(ICAM-1),干扰素诱导蛋白-10(IP-10),CC趋化因子2(CCL2),CXC趋化因子1/2/3(CXCL1/2/3),CXC趋化因子9(CXCL9/Mig)等多种炎症相关信号分子的转录表达。结论:天麻素对OGD/R损伤皮层神经细胞具有保护作用,其机制主要与抑制TLR-NF-κB-TNF炎症级联相关信号通路表达相关。Objective ： To study the protective effect of gastrodin on primary cortical neurons damaged by oxygen-glucose deprivation and reperfusion （OGD/R）, and the impact on expression of inflammation-related signaling pathway. Method： The primary cortical neurons of rats were randomly divided into control group, model group, and high, middle and low-dose gastrodin （80, 60, 40 mg·L-1） . The OGD/R damage model was built by oxygen-glucose deprivation and reperfusion. The survival rate of nerve cells was determinated by 3- （4, 5-dimethyl-2-thiazolyl） -2, 5-diphengl-2-H-tetrazolium bromide （MTT） method, and the leakage rate of lactate dehydrogenase （LDH） was measured by colorimetric method, high-throughput RNA-Seq transcriptome sequencing analysis was used to detect the differential expressions of all genes in the treated cells, and the OGD/R damage or protection-related genes or/and metabolic pathways were screened by functional annotation and enrichment analysis. Result： The activity of cortical cells decreased and the release of LDH increased after OGD/R, while gastrodin pretreatment increased cell viability and significantly lowered the LDH leakage rate. The sequence analysis revealed that the molecular mechanism mainly involved the inhibition of NF-kappa B signaling pathway （NF-KB, 18 gene）, and tumor necrosis factor signaling pathway （TNF, 21 gene）. Compared with the model group, gastrodin inhibited expressions of NF-KB, Toll like receptor 2 （TLR2）, tumor necrosis factor receptor 1/2 （TNFR1/2） , interleukin-6 （ IL-6 ） , intercellular cell adhesion molecule-1 （ ICAM-1 ） , interferon-inducible protein-lO （IP-10）, monocyte chemoattractant protein-1 （CCL2/MCP-1）, CXC-chemokine 1/2/3 （CXCL1/2/ 3）, CXC-chemokine 9 （CXCL9/Mig）, and other inflammatory signaling molecules. Conclusion： Gastrodin has a protective effect on the cortical neurons of rats after oxygen-glucose deprivation and reperfusion, and its mechanism is mainly related to the inhibition of TLR-NF

关 键 词：天麻素 皮层神经细胞 糖氧剥夺再复供 转录组 炎症 

分 类 号：R285.5[医药卫生—中药学]