Department of Orthopaedics, The Affiliated Weihai Second Hospital of Qingdao University, Weihai 264200, China
Department of Orthopaedics, The Affiliated Weihai Second Hospital of Qingdao University, Weihai 264200, China.
Biosci Rep. 2019 Mar 26;39(3). doi: 10.1042/BSR20190043. Print 2019 Mar 29.
Nucleus pulposus (NP) cell apoptosis is a classical cellular character during intervertebral disc degeneration (IDD). Previous studies have shown that inflammatory cytokine-induced NP cell apoptosis plays an important role in disc degeneration. The present study was aimed to investigate whether resveratrol can suppress IL-1β-mediated NP cell apoptosis and the potential signal transduction pathway. Experimental rat NP cells were treated with culture medium containing IL-1β (20 ng/ml) for 7 days. Control NP cells were cultured in the baseline medium. Resveratrol was added along with culture medium to investigate its effects. The inhibitor LY294002 was used to study the role of the PI3K/Akt pathway. NP cell apoptosis was reflected by the caspase-3 activity, cell apoptosis ratio, and expression of apoptosis-related molecules (Bcl-2, Bax, caspase-3, cleaved caspase-3, and cleaved PARP). Compared with the control NP cells, IL-1β significantly increased caspase-3 activity, NP cell apoptosis ratio and mRNA/protein expression of Bax, caspase-3, cleaved caspase-3 and cleaved PARP, but decreased mRNA expression of Bcl-2. However, resveratrol partly suppressed the effects of IL-1β on those cell apoptosis-related parameters. Further analysis showed that IL-1β significantly decreased activity of the PI3K/Akt pathway whereas resveratrol partly increased activity of the PI3K/Akt pathway in NP cells treated with IL-1β. Additionally, when the inhibitor LY294002 was added along with the resveratrol, its protective effects against IL-1β-induced NP cell apoptosis were attenuated. In conclusion, resveratrol suppresses IL-1β-mediated NP cell apoptosis through activating the PI3K/Akt pathway. Resveratrol may be an effective drug to attenuate inflammatory cytokine-induced disc degenerative changes.
核髓细胞（NP）凋亡是椎间盘退变（IDD）过程中的一个经典的细胞特征。先前的研究表明，炎性细胞因子诱导的 NP 细胞凋亡在椎间盘退变中起着重要作用。本研究旨在探讨白藜芦醇是否可以抑制白细胞介素-1β（IL-1β）介导的 NP 细胞凋亡及其潜在的信号转导途径。将实验用大鼠 NP 细胞用含有 20ng/mlIL-1β的培养基培养 7 天。对照组 NP 细胞用基础培养基培养。同时用培养基加入白藜芦醇，以观察其作用。用 PI3K/Akt 通路抑制剂 LY294002 研究该通路的作用。通过 caspase-3 活性、细胞凋亡率以及凋亡相关分子（Bcl-2、Bax、caspase-3、cleaved caspase-3 和 cleaved PARP）的表达来反映 NP 细胞凋亡。与对照组 NP 细胞相比，IL-1β显著增加了 caspase-3 活性、NP 细胞凋亡率以及 Bax、caspase-3、cleaved caspase-3 和 cleaved PARP 的 mRNA/蛋白表达，但降低了 Bcl-2 的 mRNA 表达。然而，白藜芦醇部分抑制了 IL-1β对这些细胞凋亡相关参数的作用。进一步分析表明，IL-1β显著降低了 NP 细胞中 PI3K/Akt 通路的活性，而白藜芦醇部分增加了 IL-1β处理的 NP 细胞中 PI3K/Akt 通路的活性。此外，当抑制剂 LY294002 与白藜芦醇一起加入时，其对 IL-1β诱导的 NP 细胞凋亡的保护作用减弱。综上所述，白藜芦醇通过激活 PI3K/Akt 通路抑制 IL-1β介导的 NP 细胞凋亡。白藜芦醇可能是一种减轻炎性细胞因子诱导的椎间盘退行性变化的有效药物。
