Rodriguez Juan L, Lopez Joseph A, Steel J Jordan
Biology Department, Colorado State University-Pueblo, 2200 Bonforte Blvd LS220, Pueblo, CO, 81001, USA.
Department of Biology, US Air Force Academy, 2355 Faculty Dr. DFB, Colorado Springs, CO, USA.
J Cannabis Res. 2021 Apr 23;3(1):10. doi: 10.1186/s42238-021-00068-y.
Sindbis virus (Alphaviridae) is a plus-strand RNA virus that is dependent on the host cell for replication. Cannabinoid (CB) receptors are found on most human cells, including virally infected cells. Activation of cannabinoid receptors has been shown to alter normal cellular physiology. This study aimed to assess how agonist (ACEA) or antagonists/inverse agonist (AM251) of the cannabinoid receptors would alter the cellular environment and impact Sindbis virus replication.
Human hepatoma (Huh7) cells were used as our model for viral replication. Cells were infected with Sindbis virus (SINV) and then treated with CB agonist (ACEA) (10 μM) or antagonist/inverse agonist (AM-251) (10 μM) and virus replication was monitored. A double subgenomic Sindbis virus containing a green fluorescent protein (GFP) reporter gene inserted into a 3' subgenomic promoter was utilized for these assays to quickly measure viral replication. GFP fluorescent cells were analyzed using flow cytometry to measure the percentage of cells expressing the viral reporter and also quantify the levels of GFP fluorescence.
Treatment of SINV-infected Huh7 cells with CB1 receptor antagonist/inverse agonist (AM251, 10 μM) resulted in a significant decrease in viral replication, while infected cells treated with a CB1 receptor agonist (ACEA, 10 μM) resulted in a significant increase of viral infection. The data indicates that activation of CB1 receptor by cannabinoids significantly influences the ability of Sindbis virus to replicate in the host cell.
Blocking CB1 receptor activity with 10 μM AM251 reduced viral replication, but activating the CB1 receptor with 10 μM ACEA resulted in an increase in viral infection. These results indicate cannabinoids may significantly impact a virus replicating in human liver cells. Future confirmation with other viruses and cell lines will be performed to better understand the impact of cannabinoids on viral infections.
辛德毕斯病毒（披膜病毒科）是一种正链RNA病毒，其复制依赖于宿主细胞。大麻素（CB）受体存在于大多数人类细胞上，包括被病毒感染的细胞。已证明大麻素受体的激活会改变正常细胞生理。本研究旨在评估大麻素受体的激动剂（ACEA）或拮抗剂/反向激动剂（AM251）如何改变细胞环境并影响辛德毕斯病毒的复制。
将人肝癌（Huh7）细胞用作病毒复制模型。用辛德毕斯病毒（SINV）感染细胞，然后用CB激动剂（ACEA）（10 μM）或拮抗剂/反向激动剂（AM - 251）（10 μM）处理，并监测病毒复制。使用一种在3'亚基因组启动子中插入了绿色荧光蛋白（GFP）报告基因的双亚基因组辛德毕斯病毒进行这些试验，以快速测量病毒复制。使用流式细胞术分析GFP荧光细胞，以测量表达病毒报告基因的细胞百分比，并量化GFP荧光水平。
用CB1受体拮抗剂/反向激动剂（AM251，10 μM）处理SINV感染的Huh7细胞导致病毒复制显著减少，而用CB1受体激动剂（ACEA，10 μM）处理感染细胞导致病毒感染显著增加。数据表明大麻素对CB1受体的激活显著影响辛德毕斯病毒在宿主细胞中的复制能力。
用10 μM AM251阻断CB1受体活性可减少病毒复制，但用10 μM ACEA激活CB1受体导致病毒感染增加。这些结果表明大麻素可能显著影响在人肝细胞中复制病毒。未来将用其他病毒和细胞系进行验证，以更好地了解大麻素对病毒感染的影响。
