Abstract

Curcumin (Cur) is a flavonoid derived from Curcuma longa L. that has been shown to have a variety of biological activities, but some previous studies have described its non-negligible negative effects on female reproduction and embryo development. To further explore the toxic stress effect, this study investigated apoptosis and autophagy of healthy buffalo (Bubalus bubalis) derived granulosa cells (GCs) exposed to Cur and/or autophagy inhibitors. Results showed that Cur declined viability of GCs in a concentration-dependent manner. Apoptosis was observed in Cur-treated GCs from 3 h. Meanwhile, under Cur stress, autophagosomes accumulated in cells, and the expression levels of autophagy key proteins LC3 and Beclin 1 were up-regulated, suggesting that Cur could induce autophagy in GCs. Early autophagy inhibitor 3-methyladenine (3-MA) increased the apoptosis rate of Cur exposed GCs, but the autophagosome degradation inhibitor chloroquine (CQ) had no effect on the apoptosis rate. The network pharmacological and molecular docking analysis indicated that the perturbation of IKK/NF-κB might be the cause of Cur toxicity toward GCs. This study unveiled another side of Cur pharmacological effects that programmed cell death can be induced by Cur in GCs, suggesting that it should be prudent to use Cur as a clinical drug for its side effects on the female reproductive system.

摘要

姜黄素 (Cur) 是一种从姜黄中提取的类黄酮，已被证明具有多种生物活性，但之前的一些研究已经描述了其对雌性生殖和胚胎发育的不可忽视的负面影响。为了进一步探索毒性应激效应，本研究调查了健康水牛（Bubalus bubalis ）的细胞凋亡和自噬。) 衍生的颗粒细胞 (GC) 暴露于 Cur 和/或自噬抑制剂。结果表明，Cur 以浓度依赖性方式降低了 GC 的活力。从 3 小时开始，在 Cur 处理的 GC 中观察到细胞凋亡。同时，在Cur胁迫下，自噬体在细胞内积累，自噬关键蛋白LC3和Beclin 1的表达水平上调，提示Cur可以诱导GCs发生自噬。早期自噬抑制剂 3-甲基腺嘌呤 (3-MA) 增加了 Cur 暴露的 GCs 的细胞凋亡率，但自噬体降解抑制剂氯喹 (CQ) 对细胞凋亡率没有影响。网络药理学和分子对接分析表明，IKK/NF-κB的扰动可能是Cur对GCs毒性的原因。