Guan Haiyu, Xie Lifen, Ji Zhenzhen, Song Rui, Qi Jieying, Nie Xiaoli
Department of Nephrology, Integrated Hospital of Traditional Chinese Medicine and Western Medicine, Southern Medical University, Guangzhou, China.
Department of Rheumatology, The Third Affiliated Hospital of Southern Medical University, Guangzhou, China.
Ann Transl Med. 2021 Sep;9(17):1384. doi: 10.21037/atm-21-3522.
Triptolide (PG490), as a triterpene dicyclic oxide has been reported to increase the generation of reactive oxygen species (ROS) and nitric oxide (NO) and induce apoptosis of RAW 264.7 cells in a dose-dependent manner. The activity of death NETs plays an important role in anti-bacterial processes in the human body. This study aimed to investigate the effect of triptolide (PG490) on neutrophil extracellular traps (NETs) formation.
After isolating peripheral blood neutrophils from healthy volunteers, cells were incubated with PG490 to observe and detect the level of NETs and detect the level of reactive oxygen species (ROS). The cells were cultured, stained and analyzed by fluorescence microscopy.
Compared with the 12-myristate-13-acetate (PMA) group, the average fluorescence intensity of SYTOX Green in the PG490 + PMA group, as detected by a multifunctional microplate reader, was significantly decreased. Intracellular ROS were labeled by fluorescence, with fluorescence intensity then measured by multifunctional microplate reader and flow cytometry. The results showed that compared with the control group, the fluorescence intensity of the PMA group was significantly increased, while there was no significant difference between PMA group and PG490 + PMA group.
The production of NETs is inhibited by PG490 , which is not associated with the level of cellular ROS. This suggests that PG490in Tripterygium wilfordii Hook F can suppress related diseases.
雷公藤内酯醇(PG490)作为一种三萜二环氧化物,据报道可增加活性氧(ROS)和一氧化氮(NO)的生成,并以剂量依赖的方式诱导RAW 264.7细胞凋亡。死亡网的活性在人体抗菌过程中起重要作用。本研究旨在探讨雷公藤内酯醇(PG490)对中性粒细胞胞外诱捕网(NETs)形成的影响。
从健康志愿者中分离外周血中性粒细胞后,将细胞与PG490孵育,以观察和检测NETs水平并检测活性氧(ROS)水平。通过荧光显微镜对细胞进行培养、染色和分析。
与12-肉豆蔻酸酯-13-乙酸酯(PMA)组相比,多功能酶标仪检测PG490 + PMA组中SYTOX Green的平均荧光强度显著降低。用荧光标记细胞内ROS,然后通过多功能酶标仪和流式细胞术测量荧光强度。结果显示,与对照组相比,PMA组的荧光强度显著增加,而PMA组与PG490 + PMA组之间无显著差异。
PG490抑制NETs的产生,这与细胞ROS水平无关。这表明雷公藤中的PG490可抑制相关疾病。