中药方剂解热 1 号抑制单纯疱疹病毒 2 感染诱导的 caspase-1 依赖性细胞焦亡。
The Chinese herbal prescription JieZe-1 inhibits caspase-1-dependent pyroptosis induced by herpes simplex virus-2 infection in vitro.
机构信息
Institute of Integrated Traditional Chinese and Western Medicine, Department of Traditional Chinese Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China.
Institute of Integrated Traditional Chinese and Western Medicine, Department of Traditional Chinese Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China.
出版信息
J Integr Med. 2023 May;21(3):277-288. doi: 10.1016/j.joim.2023.03.003. Epub 2023 Mar 9.
OBJECTIVE
JieZe-1 (JZ-1), a Chinese herbal prescription, has an obvious effect on genital herpes, which is mainly caused by herpes simplex virus type 2 (HSV-2). Our study aimed to address whether HSV-2 induces pyroptosis of VK2/E6E7 cells and to investigate the anti-HSV-2 activity of JZ-1 and the effect of JZ-1 on caspase-1-dependent pyroptosis.
METHODS
HSV-2-infected VK2/E6E7 cells and culture supernate were harvested at different time points after the infection. Cells were co-treated with HSV-2 and penciclovir (0.078125 mg/mL) or caspase-1 inhibitor VX-765 (24 h pretreatment with 100 μmol/L) or JZ-1 (0.078125-50 mg/mL). Cell counting kit-8 assay and viral load analysis were used to evaluate the antiviral activity of JZ-1. Inflammasome activation and pyroptosis of VK2/E6E7 cells were analyzed using microscopy, Hoechst 33342/propidium iodide staining, lactate dehydrogenase release assay, gene and protein expression, co-immunoprecipitation, immunofluorescence, and enzyme-linked immunosorbent assay.
RESULTS
HSV-2 induced pyroptosis of VK2/E6E7 cells, with the most significant increase observed 24 h after the infection. JZ-1 effectively inhibited HSV-2 (the 50% inhibitory concentration = 1.709 mg/mL), with the 6.25 mg/mL dose showing the highest efficacy (95.76%). JZ-1 (6.25 mg/mL) suppressed pyroptosis of VK2/E6E7 cells. It downregulated the inflammasome activation and pyroptosis via inhibiting the expression of nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing protein 3 (P < 0.001) and interferon-γ-inducible protein 16 (P < 0.001), and their interactions with apoptosis-associated speck-like protein containing a caspase recruitment domain, and reducing cleaved caspase-1 p20 (P < 0.01), gasdermin D-N (P < 0.01), interleukin (IL)-1β (P < 0.001), and IL-18 levels (P < 0.001).
CONCLUSION
JZ-1 exerts an excellent anti-HSV-2 effect in VK2/E6E7 cells, and it inhibits caspase-1-dependent pyroptosis induced by HSV-2 infection. These data enrich our understanding of the pathologic basis of HSV-2 infection and provide experimental evidence for the anti-HSV-2 activity of JZ-1. Please cite this article as: Liu T, Shao QQ, Wang WJ, Liu TL, Jin XM, Xu LJ, Huang GY, Chen Z. The Chinese herbal prescription JieZe-1 inhibits caspase-1-dependent pyroptosis induced by herpes simplex virus-2 infection in vitro. J Integr Med. 2023; 21(3): 277-288.
目的
中药方剂结泽-1（JZ-1）对单纯疱疹病毒 2 型（HSV-2）引起的生殖器疱疹有明显疗效。本研究旨在探讨 HSV-2 是否诱导 VK2/E6E7 细胞发生细胞焦亡，以及 JZ-1 的抗 HSV-2 活性和对 JZ-1 诱导的 caspase-1 依赖性细胞焦亡的影响。
方法
在感染后不同时间点采集 HSV-2 感染的 VK2/E6E7 细胞和培养上清液。细胞用 HSV-2 和喷昔洛韦（0.078125 mg/mL）或 caspase-1 抑制剂 VX-765（100 μmol/L 预处理 24 h）或 JZ-1（0.078125-50 mg/mL）共处理。用细胞计数试剂盒-8 测定和病毒载量分析评价 JZ-1 的抗病毒活性。通过显微镜、Hoechst 33342/碘化丙啶染色、乳酸脱氢酶释放测定、基因和蛋白表达、共免疫沉淀、免疫荧光和酶联免疫吸附试验分析 VK2/E6E7 细胞的炎性小体激活和细胞焦亡。
结果
HSV-2 诱导 VK2/E6E7 细胞发生细胞焦亡，感染后 24 h 时最为明显。JZ-1 能有效抑制 HSV-2（半数抑制浓度=1.709 mg/mL），6.25 mg/mL 剂量的效果最佳（95.76%）。JZ-1（6.25 mg/mL）抑制 VK2/E6E7 细胞的细胞焦亡。它通过抑制核苷酸结合寡聚化结构域样受体家族含pyrin 结构域蛋白 3（P < 0.001）和干扰素-γ诱导蛋白 16（P < 0.001）的表达，及其与凋亡相关斑点样蛋白含有 caspase 募集结构域的相互作用，降低了裂解的 caspase-1 p20（P < 0.01）、gasdermin D-N（P < 0.01）、白细胞介素（IL）-1β（P < 0.001）和 IL-18 水平（P < 0.001），从而抑制炎性小体激活和细胞焦亡。
结论
JZ-1 在 VK2/E6E7 细胞中具有良好的抗 HSV-2 作用，抑制 HSV-2 感染诱导的 caspase-1 依赖性细胞焦亡。这些数据丰富了我们对 HSV-2 感染病理基础的认识，为 JZ-1 的抗 HSV-2 活性提供了实验依据。请引用本文：Liu T, Shao QQ, Wang WJ, Liu TL, Jin XM, Xu LJ, Huang GY, Chen Z. The Chinese herbal prescription JieZe-1 inhibits caspase-1-dependent pyroptosis induced by herpes simplex virus-2 infection in vitro. J Integr Med. 2023; 21(3): 277-288.
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