绿色纳米银颗粒对正常和肝癌细胞体外细胞凋亡和氧化应激的影响。

Effects of Green Silver Nanoparticles on Apoptosis and Oxidative Stress in Normal and Cancerous Human Hepatic Cells in vitro.

机构信息

Department of Biology, College of Science, Princess Nourah Bint Abdulrahman University, Riyadh, Saudi Arabia.

Department of Zoology, College of Science, King Saud University, Riyadh, Saudi Arabia.

出版信息

Int J Nanomedicine. 2020 Mar 9;15:1537-1548. doi: 10.2147/IJN.S239861. eCollection 2020.

INTRODUCTION

Extensive use of metallic nanomaterials in different areas of agriculture and commercial products induce significant harmful effects on human health and the environment. In the current study, we synthesized an eco-friendly approach silver nanoparticles (AgNPs) using root extracts of L.

METHODS

The synthesized green silver nanoparticles (gAgNPs) were characterized by dynamic light scattering (DLS) and high-resolution transmission electron microscope (HR-TEM). The gAgNPs had a round shape and the mean size was 20-50 nm. The cytotoxic effects of gAgNPs were determined in human hepatic normal (CHANG) and cancer (HUH-7) cells by using tetrazolium salt (MTT) and lactate dehydrogenase (LDH) assays for 24 h.

RESULTS AND DISCUSSION

It was clear from the observations of this experiment that higher concentrations of gAgNPs reduce cell viability. The production of reactive oxygen species (ROS) was evaluated by using DCFDA. The gAgNPs induced more ROS in the HuH-7 cells than in the CHANG cells. The fragmentation of DNA was evaluated by alkaline single-cell gel electrophoresis and the maximum DNA strand breakage was found at a higher concentration exposure of gAgNPs for 24 h. It is important to notice that the HuH-7 cells showed an increased sensitivity to gAgNPs than the CHANG cells. The apoptotic and necrotic effects of gAgNPs on both the cells were evaluated using annexin-V-FITC and propidium iodide staining. An increased count of apoptotic and necrotic cells was found following a higher concentration exposure of gAgNPs. The apoptotic protein expression in these cells due to gAgNPs exposure was determined using immunoblotting techniques and the level of Bcl2 was decreased. However, the expression of BAX and protein was increased in both cells.

CONCLUSION

Therefore, it can be concluded that higher concentrations of gAgNPs may induce significant cytotoxicity and cause DNA damage and apoptosis.

简介

在农业和商业产品的不同领域中广泛使用金属纳米材料会对人类健康和环境产生重大的有害影响。在本研究中，我们使用 L. 的根提取物合成了一种环保的银纳米粒子（AgNPs）。

方法

通过动态光散射（DLS）和高分辨率透射电子显微镜（HR-TEM）对合成的绿色银纳米粒子（gAgNPs）进行了表征。gAgNPs 呈圆形，平均尺寸为 20-50nm。通过噻唑蓝（MTT）和乳酸脱氢酶（LDH）测定法在人肝正常（CHANG）和肝癌（HUH-7）细胞中测定 gAgNPs 的细胞毒性作用，持续 24 小时。

结果与讨论

从实验观察结果清楚地看出，较高浓度的 gAgNPs 会降低细胞活力。通过 DCFDA 评估活性氧（ROS）的产生。gAgNPs 在 HuH-7 细胞中诱导的 ROS 比在 CHANG 细胞中多。通过碱性单细胞凝胶电泳评估 DNA 的片段化，发现 gAgNPs 暴露于更高浓度时，最大 DNA 链断裂。重要的是要注意，与 CHANG 细胞相比，HuH-7 细胞对 gAgNPs 更为敏感。通过 Annexin-V-FITC 和碘化丙啶染色评估 gAgNPs 对两种细胞的凋亡和坏死作用。发现 gAgNPs 暴露于更高浓度时，凋亡和坏死细胞的计数增加。通过免疫印迹技术确定了这些细胞中由于 gAgNPs 暴露而导致的凋亡蛋白表达，发现 Bcl2 水平降低。然而，BAX 和蛋白的表达在两种细胞中均增加。