Zhang Xiaoyan, Hao Jinfang, Sun Chenxi, Du Jianping, Han Qian, Li Qingshan
Department of Laboratory Medicine of Fenyang College, School of Pharmaceutical Science, Shanxi Medical University, Taiyuan, Shanxi 030001, P.R. China.
Shanxi Key Laboratory of Innovative Drugs for The Treatment of Serious Diseases Based on Chronic Inflammation, Shanxi University of Chinese Medicine, Jinzhong, Shanxi 030619, P.R. China.
Exp Ther Med. 2022 Mar;23(3):237. doi: 10.3892/etm.2022.11162. Epub 2022 Jan 24.
Enterovirus 71 (EV71) is one of the primary pathogens involved in severe hand, foot and mouth disease in children. EV71 infection causes various types of programmed cell death. However, there are currently no clinically approved specific antiviral drugs for control of EV71 infection. (AM), a Traditional Chinese medicine, has been used in antiviral therapy in China. The aim of the present study was to determine whether total astragalosides (ASTs), bioactive components of AM, protect against EV. DAPI nuclear staining was used to observe morphological changes of the nucleus and the protective effect of ASTs, which revealed that the nucleus shrank following EV71 infection, while ASTs reversed it. Cell Counting Kit-8 assay found that human normal gastric epithelial cell (GES-1 cell) viability decreased following EV71 infection, while lactate dehydrogenase (LDH) assay showed that EV71 infection induced GES-1 cell damage. Western blotting was used to measure the expression levels of apoptosis and pyroptosis marker protein to determine whether EV71 infection induced apoptosis and pyroptosis in GES-1 cells. Reverse transcription-quantitative PCR was used to determine the anti-EV71 effect of ASTs. The results showed that ASTs protected GES-1 cells from EV71-induced cell apoptosis and pyroptosis. Furthermore, the present data demonstrated that the protective effect of ASTs was exerted by suppressing EV71 replication and release. These findings suggested that ASTs may represent a potential antiviral agent for the treatment of EV71 infection.
肠道病毒71型(EV71)是导致儿童重症手足口病的主要病原体之一。EV71感染可引发多种类型的程序性细胞死亡。然而,目前尚无临床批准的用于控制EV71感染的特异性抗病毒药物。黄芪(AM)是一种中药,已在中国用于抗病毒治疗。本研究的目的是确定黄芪的生物活性成分总黄芪苷(ASTs)是否对EV71有保护作用。采用4',6-二脒基-2-苯基吲哚(DAPI)核染色观察细胞核的形态变化及ASTs的保护作用,结果显示EV71感染后细胞核缩小,而ASTs可使其恢复。细胞计数试剂盒-8(CCK-8)检测发现,EV71感染后人正常胃上皮细胞(GES-1细胞)活力下降,而乳酸脱氢酶(LDH)检测表明EV71感染可诱导GES-1细胞损伤。采用蛋白质免疫印迹法检测凋亡和焦亡标志物蛋白的表达水平,以确定EV71感染是否诱导GES-1细胞发生凋亡和焦亡。采用逆转录-定量聚合酶链反应(RT-qPCR)检测ASTs的抗EV71作用。结果表明,ASTs可保护GES-1细胞免受EV71诱导的细胞凋亡和焦亡。此外,本研究数据表明,ASTs的保护作用是通过抑制EV71的复制和释放来实现的。这些发现提示,ASTs可能是一种治疗EV71感染的潜在抗病毒药物。