Chen Haojie, Tang Guoke, Yu Jiangming, Yu Ronghua
Department of Orthopedics, Tongren Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200336, China.
Department of Orthopedics, Shanghai General Hospital, Shanghai Jiaotong University, No. 100 Haining Road, Shanghai 200080, China.
Evid Based Complement Alternat Med. 2022 Jul 12;2022:1731701. doi: 10.1155/2022/1731701. eCollection 2022.
Curcumin (Cur) is a natural polyphenol isolated from turmeric and has potent anti-inflammatory and antioxidant activities. This study aimed to explore the effects and possible mechanisms of curcumin on oxygen-glucose deprivation/reoxygenation (OGD/R)-induced injury in cultured rat astrocyte primary cells.
After screening for effective doses, the cultured rat astrocyte primary cells were divided into three groups: control, OGD/R, and OGD/R + curcumin (10 M, 20 M, and 40 M). Cell viability was detected using CCK8 assays. The level of malondialdehyde and superoxide dismutase activity was determined using commercial kits. The endothelial nitric oxide synthase and adenosine triphosphate concentrations were determined by enzyme-linked immunosorbent assay. The mRNA levels of the inflammatory indexes interleukin (IL)-6, tumor necrosis factor (TNF)-alpha, and interleukin (IL)-1 were evaluated by quantitative reverse-transcription polymerase chain reaction. Annexin V-fluorescein isothiocyanate/propidium iodide was used to detect apoptosis. JC-1 was used to assess the mitochondrial membrane potential. The protein expression of apoptosis-related proteins (B-cell lymphoma-2 (Bcl-2), BCL-2-associated X (Bax), and cleaved caspase 3), mitochondria-related proteins (dynamin-related protein 1 (DRP1), phosphorylated DRP1 (p-DRP1), and mitofusin 2), and essential proteins of the extracellular signal-regulated kinase (ERK) signaling pathway (ERK1/2, p-ERK1/2) were analyzed by western blot.
Our data indicated that curcumin reversed OGD/R-induced cell viability loss, oxidative stress, inflammatory cytokine production, and cell apoptosis in a dose-dependent manner. Furthermore, curcumin attenuated OGD/R-induced mitochondrial dysfunction and ERK1/2 phosphorylation in a dose-dependent manner.
Curcumin protected against OGD/R-induced injury in rat astrocyte primary cells through improving mitochondrial function and regulating the ERK signaling pathway.
姜黄素(Cur)是从姜黄中分离出的一种天然多酚,具有强大的抗炎和抗氧化活性。本研究旨在探讨姜黄素对氧糖剥夺/复氧(OGD/R)诱导的原代培养大鼠星形胶质细胞损伤的影响及可能机制。
筛选出有效剂量后,将原代培养的大鼠星形胶质细胞分为三组:对照组、OGD/R组、OGD/R + 姜黄素组(10 μM、20 μM和40 μM)。采用CCK8法检测细胞活力。使用商业试剂盒测定丙二醛水平和超氧化物歧化酶活性。通过酶联免疫吸附测定法测定内皮型一氧化氮合酶和三磷酸腺苷浓度。采用定量逆转录聚合酶链反应评估炎症指标白细胞介素(IL)-6、肿瘤坏死因子(TNF)-α和白细胞介素(IL)-1的mRNA水平。使用膜联蛋白V-异硫氰酸荧光素/碘化丙啶检测细胞凋亡。使用JC-1评估线粒体膜电位。通过蛋白质印迹法分析凋亡相关蛋白(B细胞淋巴瘤-2(Bcl-2)、BCL-2相关X蛋白(Bax)和裂解的半胱天冬酶3)、线粒体相关蛋白(动力相关蛋白1(DRP1)、磷酸化DRP1(p-DRP1)和线粒体融合蛋白2)以及细胞外信号调节激酶(ERK)信号通路的关键蛋白(ERK1/2、p-ERK1/2)的蛋白表达。
我们的数据表明,姜黄素以剂量依赖性方式逆转了OGD/R诱导的细胞活力丧失、氧化应激、炎性细胞因子产生和细胞凋亡。此外,姜黄素以剂量依赖性方式减轻了OGD/R诱导的线粒体功能障碍和ERK1/2磷酸化。
姜黄素通过改善线粒体功能和调节ERK信号通路,保护原代培养大鼠星形胶质细胞免受OGD/R诱导的损伤。