Bergenin is a C-glucoside of 4-O-methyl gallic acid with a variety of biological activities, such as antioxidant and anti-inflammatory. Herein, we investigated the involvement of bergenin in the protective effect against H₂O₂-induced oxidative stress and apoptosis in human nucleus pulposus cells (HNPCs) and the underlying mechanisms. HNPCs were cotreated with various concentrations of bergenin and 200 μM H₂O₂ for 24 h. Cell viability was detected by Cell Counting Kit-8 and lactate dehydrogenase release assays. Reactive oxygen species (ROS) was evaluated utilizing 2′,7′-dichlorofluorescein-diacetate. Superoxide dismutase (SOD) and catalase (CAT) activities and glutathione (GSH) levels were measured to assess oxidative stress. Apoptosis was evaluated using terminal deoxynucleotidyl transferase dUTP nick end labeling and caspase-3/7 activity assays. Expression of protein was determined by western blotting. Results indicated that treatment with bergenin significantly alleviated H₂O₂-induced viability reduction and ROS overproduction in HNPCs in a dose-dependent manner. Bergenin alleviated H₂O₂-induced oxidative stress in HNPCs by increased activity of superoxide dismutase and level of glutathione peroxidase. H₂O₂-induced apoptosis and activity of caspase-3/7 were also suppressed by bergenin treatment in HNPCs. Western blotting showed that H₂O₂-induced decrease in expression of peroxisome proliferator-activated receptor γ (PPAR-γ) and increase in nuclear factor κB (NF-κB) were inhibited by bergenin. However, the inhibitory effect of bergenin on H₂O₂-induced viability reduction, oxidative stress and apoptosis were noticeably abrogated in PPAR-γ knockdown HNPCs. In conclusion, our results indicated that bergenin alleviates H₂O₂-induced oxidative stress and apoptosis in HNPCs by activating PPAR-γ and suppressing NF-κB pathway.

中文翻译：

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Bergenin 减轻 H2O2 诱导的髓核细胞氧化应激和细胞凋亡：PPAR-γ/NF-κB 通路的参与

Bergenin 是 4- O-甲基没食子酸的 C-葡萄糖苷，具有多种生物活性，如抗氧化和抗炎。在此，我们研究了 bergenin 对 H ₂ O ₂诱导的人髓核细胞 (HNPCs) 氧化应激和细胞凋亡的保护作用及其潜在机制。HNPCs 用不同浓度的 bergenin 和 200 μM H ₂ O _{2 共同处理}24 小时。通过 Cell Counting Kit-8 和乳酸脱氢酶释放试验检测细胞活力。使用 2',7'-二氯荧光素-二乙酸盐评估活性氧 (ROS)。测量超氧化物歧化酶 (SOD) 和过氧化氢酶 (CAT) 活性以及谷胱甘肽 (GSH) 水平以评估氧化应激。使用末端脱氧核苷酸转移酶 dUTP 缺口末端标记和 caspase-3/7 活性测定评估细胞凋亡。通过蛋白质印迹确定蛋白质的表达。结果表明，用bergenin处理以剂量依赖性方式显着减轻H ₂ O ₂诱导的HNPC活力降低和ROS过量产生。Bergenin 减轻 H ₂ O ₂-通过增加超氧化物歧化酶的活性和谷胱甘肽过氧化物酶的水平来诱导 HNPC 的氧化应激。H ₂ O ₂诱导的细胞凋亡和 caspase-3/7 的活性也被 HNPC 中的 bergenin 处理所抑制。蛋白质印迹表明，H ₂ O ₂诱导的过氧化物酶体增殖物激活受体γ (PPAR-γ) 表达的降低和核因子κB (NF-κB) 的增加被bergenin 抑制。然而，在 PPAR-γ 敲低的 HNPCs 中，bergenin对 H ₂ O ₂诱导的活力降低、氧化应激和细胞凋亡的抑制作用明显消失。总之，我们的结果表明，bergenin 减轻了 H ₂ O ₂-通过激活 PPAR-γ 和抑制 NF-κB 通路诱导 HNPCs 的氧化应激和细胞凋亡。