Hu Shengzhao, Chen Yingli, Huang Shipeng, Liu Min, Liu Ying, Huang Shaofang
Department of Emergency, the First Affiliated Hospital of Nanchang University, Nanchang, China.
Department of Hematology, Jiangxi Provincial Children's Hospital, Nanchang, China.
Ann Transl Med. 2022 Oct;10(20):1097. doi: 10.21037/atm-22-2143.
Cerebral ischemic stroke is a serious condition with high incidence, mortality, and associated disability. Currently, effective therapeutic options are available for ischemic stroke are limited. Accumulating evidence indicates that sodium Danshensu, mono sodium compound derived from Salvia miltiorrhiza, plays protective roles in ischemic stroke. However, the underlying protective mechanism of sodium Danshensu in cerebral ischemic stroke remains unknown.
In the current study, we explored the role and mechanism of sodium Danshensu on astrocytes exposed to oxygen-glucose deprivation/reoxygenation (OGD/R), which mimics the process of ischemia-reperfusion. The impact of sodium Danshensu on cell viability and apoptosis after OGD/R were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-dophenyl tetrazolium bromide (MTT) assay and flow cytometry. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot were used to detect the expression of target messenger RNA (mRNA) and proteins associated with apoptosis and autophagy. The release of lactate dehydrogenase (LDH) was determined, and the production of proinflammatory cytokines were detected using enzyme-linked immunosorbent assay (ELISA) kits.
It was found that sodium Danshensu could significantly increase cell viability and decrease LDH release and apoptosis. Besides, it inhibited the production of proinflammatory cytokines, including tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, and IL-6. Sodium Danshensu also dose-dependently decreased protein and mRNA levels of nucleotide binding oligomerization NOD-like receptor pyrin domain containing 3 (NLRP3) and high mobility group box 1 (HMGB1), which play a crucial role in promoting ischemic stroke-induced cell injury. Moreover, sodium Danshensu dose-dependently upregulated Beclin 1 expression, downregulated P62 protein expression, and further increased LC3B-II/LC3B-I ratio through inducing autophagy in astrocytes. Additionally, we noticed that sodium Danshensu dose-dependently increased tuberous sclerosis complex-2 (TSC2) protein expression, while significantly reduced the levels of mammalian target of rapamycin (mTOR) in the presence of OGD/R insult.
These findings suggest that sodium Danshensu protects against OGD/R-induced injury by modulating the NLRP3 inflammasome and TSC2/mTOR pathways.
脑缺血性中风是一种严重的疾病，具有高发病率、高死亡率及相关残疾率。目前，针对缺血性中风的有效治疗选择有限。越来越多的证据表明，丹参素钠，一种从丹参中提取的单钠化合物，在缺血性中风中发挥保护作用。然而，丹参素钠在脑缺血性中风中的潜在保护机制仍不清楚。
在本研究中，我们探讨了丹参素钠对暴露于氧糖剥夺/复氧（OGD/R）的星形胶质细胞的作用及机制，OGD/R模拟了缺血再灌注过程。通过3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）法和流式细胞术评估丹参素钠对OGD/R后细胞活力和凋亡的影响。采用定量逆转录聚合酶链反应（qRT-PCR）和蛋白质印迹法检测与凋亡和自噬相关的靶信使核糖核酸（mRNA）和蛋白质的表达。测定乳酸脱氢酶（LDH）的释放，并使用酶联免疫吸附测定（ELISA）试剂盒检测促炎细胞因子的产生。
发现丹参素钠可显著提高细胞活力，降低LDH释放和凋亡。此外，它抑制促炎细胞因子的产生，包括肿瘤坏死因子-α（TNF-α）、白细胞介素（IL）-1β和IL-6。丹参素钠还剂量依赖性地降低核苷酸结合寡聚化结构域样受体含pyrin结构域3（NLRP3）和高迁移率族蛋白B1（HMGB1）的蛋白质和mRNA水平，它们在促进缺血性中风诱导的细胞损伤中起关键作用。此外，丹参素钠剂量依赖性地上调Beclin 1表达，下调P62蛋白表达，并通过诱导星形胶质细胞自噬进一步提高LC3B-II/LC3B-I比值。此外，我们注意到在存在OGD/R损伤的情况下，丹参素钠剂量依赖性地增加结节性硬化复合物-2（TSC2）蛋白表达，同时显著降低雷帕霉素靶蛋白（mTOR）水平。
这些发现表明，丹参素钠通过调节NLRP3炎性小体和TSC2/mTOR途径来保护细胞免受OGD/R诱导的损伤。
