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-κB,18 gene),and tumor necrosis factor signaling pathway(TNF,21 gene).Compared with the model group,gastrodin inhibited expressions of NF-κB,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-κB-TNF inflammatory cascade-related signaling pathways.
作者
文欢
张大燕
王伟
高继海
饶朝龙
彭成
WEN Huan;ZHANG Da-yan;WANG Wei;GAO Ji-hai;RAO Chao-long;PENG Cheng(State Key Laboratory and Breeding Base of Systematic Research,Development and Utilization ofChinese Medicine Resources,School of Pharmaceutical,Chengdu University of TraditionalChinese Medicine,Chengdu 611137,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
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)
关键词
天麻素
皮层神经细胞
糖氧剥夺再复供
转录组
炎症
gastrodin
cortical nerve cell
oxygen-glucose deprivation and reperfusion
transcriptome
inflammation
