Journal of Shanghai Jiao Tong University (Medical Science) ›› 2022, Vol. 42 ›› Issue (10): 1404-1412.doi: 10.3969/j.issn.1674-8115.2022.10.005

• Basic research • Previous Articles    

Effect of chloroquine phosphate on vascular endothelial cell injury induced by double-stranded RNA

ZHANG Jiaojiao(), SUN Junnan, WANG Hairong()   

  1. Department of Emergency Medicine, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China
  • Received:2022-04-29 Accepted:2022-06-18 Online:2022-10-28 Published:2022-12-02
  • Contact: WANG Hairong;


Objective ·To investigate the inhibitory effect of chloroquine phosphate (CQ) on vascular endothelial cell injury induced by double-stranded RNA (dsRNA) and its mechanism. Methods ·An in vitro model of vascular endothelial cell injury induced by dsRNA in viral sepsis was established via stimulating human umbilical vein endothelial cells (HUVECs) with polyinosinic-polycytidylic acid (poly I:C). HUVECs were randomly divided into normal control group (NC group), poly I:C group with different concentrations and CQ+poly I:C group. The cell viability of HUVECs before and after poly I:C treatment was detected by using Cell Counting Kit-8 (CCK-8). The morphological changes of HUVECs were observed by using optical microscope. The integrity of cell membrane was assessed by calculating the rate of propidium iodide (PI) positive cells by using flow cytometry. The expression level of mature proinflammatory cytokine interleukin 1β (IL-1β), and key proteins of pyroptosis were detected by Western blotting. Results ·Compared with the NC group, HUVECs treated with poly I:C showed a decreased cell viability, more expanded "Vesicles" and more ruptured cells. An increased level of IL-1β expression and positive rate of PI+ cells were also observed. The expression levels of NOD-like receptor thermal protein domain associated protein 3 (NLRP3), and cleaved caspase-1 and gasdermin D N-terminal (GSDMD-N) were increased significantly (all P<0.05) under the treatment of poly I:C and the changes were concentration-dependent. Interestingly, compared with poly I:C group, the cell viability of HUVEC pretreated by CQ was significantly increased, and a decreased IL-1β level were observed. The phenomena of cell swelling and rupture was improved, and a lower positive rate of PI+ cells was detected. The expression levels of cleaved caspase-1 and GSDMD-N of cell pyrosis were also decreased (all P<0.05). CQ pretreatment did not reduce NLRP3 expression induced by dsRNA, but it did inhibit the expression of Toll-like receptor 3 (TLR3). Conclusion ·Pyroptosis is involved in vascular endothelial cells injury induced by dsRNA, and CQ inhibits dsRNA-induced pyroptosis, which could be related to TLR3 regulation.

Key words: chloroquine phosphate (CQ), pyroptosis, double-stranded RNA (dsRNA), vascular endothelial cell

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