中药材2023，Vol.46Issue(6)：1494-1500,7.DOI:10.13863/j.issn1001-4454.2023.06.029
原花青素经ROS介导的SIRT3/HIF-1α轴抑制缺氧PANC-1细胞增殖的研究
Study on the Inhibitory Effect of Proanthocyanidins on Proliferation of Hypoxic PANC-1 Cells via ROS Mediated SIRT3/HIF-1α Axis
摘要
Abstract
Objective:To investigate the role and mechanism of proanthocyanidin inhibiting the proliferation of hypoxic pancreatic cancer PANC-1 cells.Methods:The GEPIA website was used to predict the expressions and correlation of SIRT3 and HIF-1α genes in pancreatic cancer.The physical induction method of anaerobic culture bag was used to simulate the hypoxic environment of PANC-1 cells in vivo,and divided into normoxia control group,model group,low(15 μg/mL),medium(30 μg/mL)and high(60 μg/mL)con-centration of proanthocyanidin groups.CCK-8 assay was used to detect the effect of different concentrations of proanthocyanidin on PANC-1 cells viability.Cell morphology was observed by inverted fluoressence microscope.Annexin V-FITC/PI double staining was used to detect the apoptosis level.DCFH-DA fluorescence probe was used to detect intracellular reactive oxygen species(ROS)levels.Biochemical kits were used to detect intracellular superoxide dismutase(SOD)activity and total antioxidant capacity(T-AOC).Western Blot was used to detect the expression of SIRT3 and HIF-1α protein in cancer cells.Results:The GEPIA database predicted that SIRT3 expression tended to be upregulated in pancreatic cancer,while HIF-1α expression was significantly overexpressed,and the two were positively correlated.Under the hypoxic conditions,CCK-8 experiments showed that proanthocyanidin significantly inhibited the viability of PANC-1 cells.Fluorescence in situ showed that intracellular ROS levels were significantly increased in the model group and no signif-icant apoptosis was observed.Proanthocyanidin could reverse intracellular ROS levels and induce apoptosis.The antioxidant activity de-tection showed that intracellular SOD viability was significantly decreased,while T-AOC was significantly increased in the model group.Proanthocyanidin increased SOD viability and T-AOC in PANC-1 cells in a concentration-dependent manner.Western Blot results showed that SIRT3 expression was not significantly altered in the model group and HIF-1α expression was significantly up-regulated.Proanthocyanidin significantly down-regulated the expressions of SIRT3 and HIF-1α proteins in PANC-1 cells.Conclusion:Proanthocya-nidin can inhibit the proliferation of PANC-1 pancreatic cancer cell line,and the mechanism may be related to the inhibition of the ROS-mediated SIRT3/HIF-1α signaling pathway.关键词
原花青素/胰腺癌/活性氧/缺氧/SIRT3Key words
Proanthocyanidin/Pancreatic cancer/Reactive oxygen species/Hypoxic/SIRT3分类
医药卫生引用本文复制引用
师豪,朱颖蔚,徐克群,蒋华..原花青素经ROS介导的SIRT3/HIF-1α轴抑制缺氧PANC-1细胞增殖的研究[J].中药材,2023,46(6):1494-1500,7.基金项目
国家自然科学基金项目(81902391) （81902391）
