Key Laboratory of Dunhuang Medicine and Transformation at Provincial and Ministerial Level, Gansu University of Chinese Medicine, Lanzhou 730000, China.
Provincial-Level Key Laboratory for Molecular Medicine of Major Diseases and the Prevention and Treatment with Traditional Chinese Medicine Research in Gansu Colleges and Universities, Gansu University of Chinese Medicine, Lanzhou 730000, China.
Cell Mol Biol (Noisy-le-grand). 2023 Mar 31;69(3):156-162. doi: 10.14715/cmb/2023.69.3.23.
To investigate the protective effect of Quercetin (Que) on lung epithelial cells (BEAS-2B) induced bystander effect (RIBE) after heavy ion irradiation of A549 cells. A549 cells were irradiated with 2 Gy X heavy ion rays to obtain a conditioned medium. BEAS-2B was incubated with a conditioned medium or Que. CCK-8 assay was used to screen the optimal effective concentration of Que and detect cell proliferation. Cell number was measured by cell counter and apoptosis rate was measured by flow cytometry. HMGB1 and ROS levels were measured by ELISA. Western blot was used to detect the protein expression of HMGB1, TLR4, p65, Bcl-2, Bax, Caspase3 and Cleaved Caspase3. The growth and proliferation rate of BEAS-2B decreased while the apoptosis rate increased after conditioned medium stimulation, and Que intervention inhibited this effect. The expression of HMGB1 and ROS increased after conditioned medium stimulation, and this effect was inhibited by Que intervention. In addition, the conditioned medium increased the levels of proteins of HMGB1, TLR4, p65, Bax, Caspase3 and Cleaved Caspase 3, and decreased levels of Bcl-2 protein, but Que intervention decreased the levels of HMGB1, TLR4, p65, Bax, Caspase3 and Cleaved Caspase 3proteins, and increased levels of Bcl-2 protein. The RIBE of BEAS-2B induced by irradiation of A549 is associated with HMGB1TLR4/NF-κB signaling pathway in conditioned medium inducing apoptosis by activating ROS, and Que may block RIBE-induced apoptosis by regulating HMGB1/TLR4/NF-κB pathway.
探讨槲皮素（Que）对 X 射线重离子照射 A549 细胞后肺上皮细胞（BEAS-2B）旁分泌效应（RIBE）的保护作用。用 2 Gy X 射线重离子照射 A549 细胞获得条件培养基。将 BEAS-2B 与条件培养基或 Que 孵育。用 CCK-8 法筛选 Que 的最佳有效浓度并检测细胞增殖。用细胞计数器测量细胞数，用流式细胞术测量细胞凋亡率。用 ELISA 法测量 HMGB1 和 ROS 水平。用 Western blot 检测 HMGB1、TLR4、p65、Bcl-2、Bax、Caspase3 和 Cleaved Caspase3 的蛋白表达。条件培养基刺激后 BEAS-2B 的生长和增殖率降低，凋亡率增加，而 Que 干预抑制了这种作用。条件培养基刺激后 HMGB1 和 ROS 的表达增加，而 Que 干预抑制了这种作用。此外，条件培养基增加了 HMGB1、TLR4、p65、Bax、Caspase3 和 Cleaved Caspase3 蛋白的水平，降低了 Bcl-2 蛋白的水平，但 Que 干预降低了 HMGB1、TLR4、p65、Bax、Caspase3 和 Cleaved Caspase3 蛋白的水平，增加了 Bcl-2 蛋白的水平。A549 照射诱导的 BEAS-2B 的 RIBE 与条件培养基中 HMGB1TLR4/NF-κB 信号通路有关，该通路通过激活 ROS 诱导细胞凋亡，Que 可能通过调节 HMGB1/TLR4/NF-κB 通路阻断 RIBE 诱导的细胞凋亡。
