Bao Gang, Li Chuankun, Qi Lei, Wang Ning, He Baixiang
Department of Neurosurgery, The First Affiliated Hospital of Xi'an Jiaotong University Health Science Center, Xi'an 710061, Shaanxi, PR China.
Department of Neurosurgery, The First Affiliated Hospital of Xi'an Jiaotong University Health Science Center, Xi'an 710061, Shaanxi, PR China.
Biomed Pharmacother. 2016 Dec;84:925-930. doi: 10.1016/j.biopha.2016.10.007. Epub 2016 Oct 17.
Tetrandrine (TET) is a bis-benzylisoquinoline alkaloid, which is isolated from a Chinese medicinal herb with antioxidant and anti-inflammatory activities. In this study, we investigated the effects of TET on oxygen-glucose-serum deprivation/reoxygenation (OGSD/R)-induced injury in rat spinal cord astrocytes, which mimics hypoxic/ischemic conditions in vivo. MTT and LDH assays indicated that cell viability was distinctly reduced and LDH leakage was elevated after OGSD/R exposure, which were dose-dependently reversed by pretreatment with TET (0.1, 1, 10, 20μM). Western blot analysis showed that OGSD/R exposure resulted in an enhanced expression of Bax and Caspase-3 proteins, and Bcl-2 reduction; whereas these effects were dose-dependently restored by TET pretreatment. TET pretreatment also dose-dependently inhibited the elevated Caspase-3 activity in OGSD/R-treated astrocytes. The oxidative stress status was evaluated using commercial kits, and the results demonstrated that OGSD/R exposure induced obvious oxidative stress, accompanied by elevated levels of reactive oxygen species (ROS) and malondialdehyde (MDA), and reduced superoxide dismutase (SOD) activity, which were dose-dependently restored by TET pretreatment. In addition, TET pretreatment diminished the accumulation of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6) induced by OGSD/R. Moreover, TET pretreatment dose-dependently suppressed Akt phosphorylation and nuclear factor-kappaB (NF-κB) activity augmented by OGSD/R. Similarly, both PI3K inhibitor LY294002 and NF-κB inhibitor PDTC notably attenuated OGSD/R-induced Akt phosphorylation, NF-κB activation, ROS generation, and TNF-α secretion. Taken together, these data demonstrated that TET protected against OGSD/R-induced injury in rat spinal cord astrocytes, which may be attributed to its antioxidant and anti-inflammatory activities via PI3K/AKT/NF-κB signaling pathway.
粉防己碱（TET）是一种双苄基异喹啉生物碱，从一种具有抗氧化和抗炎活性的中草药中分离得到。在本研究中，我们研究了TET对大鼠脊髓星形胶质细胞氧-葡萄糖-血清剥夺/复氧（OGSD/R）诱导损伤的影响，该损伤模拟了体内的缺氧/缺血条件。MTT和LDH测定表明，OGSD/R暴露后细胞活力明显降低，LDH泄漏增加，而TET（0.1、1、10、20μM）预处理可剂量依赖性地逆转这些变化。蛋白质印迹分析表明，OGSD/R暴露导致Bax和Caspase-3蛋白表达增强，Bcl-2减少；而TET预处理可剂量依赖性地恢复这些效应。TET预处理还剂量依赖性地抑制了OGSD/R处理的星形胶质细胞中升高的Caspase-3活性。使用商业试剂盒评估氧化应激状态，结果表明OGSD/R暴露诱导明显的氧化应激，伴有活性氧（ROS）和丙二醛（MDA）水平升高，超氧化物歧化酶（SOD）活性降低，而TET预处理可剂量依赖性地恢复这些变化。此外，TET预处理减少了OGSD/R诱导的肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）和白细胞介素-6（IL-6）的积累。此外，TET预处理剂量依赖性地抑制了OGSD/R增强的Akt磷酸化和核因子-κB（NF-κB）活性。同样，PI3K抑制剂LY294002和NF-κB抑制剂PDTC均显著减弱了OGSD/R诱导的Akt磷酸化、NF-κB激活、ROS生成和TNF-α分泌。综上所述，这些数据表明TET可保护大鼠脊髓星形胶质细胞免受OGSD/R诱导的损伤，这可能归因于其通过PI3K/AKT/NF-κB信号通路的抗氧化和抗炎活性。
