Hebei Key Laboratory of Chinese Medicine Research on Cardio-Cerebrovascular Disease, Hebei University of Chinese Medicine, Shijiazhuang 050200, China.
Department of Neurosurgery, Hebei General Hospital, Shijiazhuang, China.
Int Immunopharmacol. 2024 Apr 20;131:111862. doi: 10.1016/j.intimp.2024.111862. Epub 2024 Mar 20.
Astragaloside IV(ASⅣ), the main component of Radix Astragali, has been used to treat cerebral ischemia reperfusion injury (CIRI). However, the molecular mechanism of ASIV in CIRI needs to be further elucidated. Long non-coding RNA (lncRNA) is considered to be an important kind of regulatory molecule in CIRI. In this work, the biological effect and molecular mechanism of ASIV in CIRI through lncRNA were analyzed by using rat middle cerebral artery occlusion and reperfusion (MCAO/R) model and primary rat microglia (RM) cells oxygen and glucose deprivation/reoxygenation (OGD/R) model. The neurological deficit score was evaluated, the volume of cerebral infarction was calculated, and pyroptosis related molecules were detected by qPCR and western blot. Then, high-throughput sequencing was performed in sham and MCAO/R groups. The competitive endogenous RNA (ceRNA) networks associated with pyroptosis were constructed by functional enrichment analysis. CCK-8 detection of cell survival rate, qPCR and western blot were used to determine the specific molecular mechanism of ASⅣ through ceRNA in vitro. Results showed thatASⅣ could decrease the neurological deficit score, reduce the volume of cerebral infarction, inhibit inflammatory reaction and pyroptosis in MCAO/R model rats. Next, the ceRNA network was established, including the LOC102555978/miR-3584-5p/NLRP3 regulatory network. In vitro experiments showed that LOC102555978 promotes NLRP3 mediated pyroptosis of RM cells through sponge adsorption of miR-3584-5p, which may provide a potential therapeutic target for post-CIRI inflammation regulation. ASⅣ could inhibit pyroptosis of RM cells by down-regulating LOC102555978. LOC102555978/miR-3584-5p/NLRP3 may be the molecular mechanism of ASⅣ's CIRI protective effect.
黄芪甲苷（ASIV）是黄芪的主要成分，已用于治疗脑缺血再灌注损伤（CIRI）。然而，ASIV 在 CIRI 中的分子机制仍需进一步阐明。长链非编码 RNA（lncRNA）被认为是 CIRI 中一种重要的调节分子。在这项工作中，通过大鼠大脑中动脉闭塞再灌注（MCAO/R）模型和原代大鼠小胶质细胞（RM）细胞氧葡萄糖剥夺/再氧合（OGD/R）模型，分析了 ASIV 通过 lncRNA 对 CIRI 的生物学效应和分子机制。通过 qPCR 和 Western blot 检测神经功能缺损评分、脑梗死体积和细胞焦亡相关分子。然后，对假手术组和 MCAO/R 组进行高通量测序。通过功能富集分析构建与细胞焦亡相关的竞争性内源性 RNA（ceRNA）网络。通过 CCK-8 检测细胞存活率、qPCR 和 Western blot 确定 ASIV 通过 ceRNA 在体外的具体分子机制。结果表明，ASIV 可降低 MCAO/R 模型大鼠的神经功能缺损评分，减少脑梗死体积，抑制炎症反应和细胞焦亡。接下来，建立了 ceRNA 网络，包括 LOC102555978/miR-3584-5p/NLRP3 调控网络。体外实验表明，LOC102555978 通过海绵吸附 miR-3584-5p 促进 RM 细胞中 NLRP3 介导的细胞焦亡，这可能为 CIRI 后炎症调节提供潜在的治疗靶点。ASIV 通过下调 LOC102555978 抑制 RM 细胞的细胞焦亡。LOC102555978/miR-3584-5p/NLRP3 可能是 ASIV 治疗 CIRI 的分子机制。
