Effect of varicella-zoster virus on nucleotide-binding oligomerization domain-like receptor protein 3 inflammasome in mouse brain astrocytes
10.35541/cjd.20240075
- VernacularTitle:水痘-带状疱疹病毒对小鼠脑星形胶质细胞内NLRP3炎症小体的作用
- Author:
Hongyan FENG
1
;
Xingwang WANG
;
Yang ZHAO
;
Huilan YANG
Author Information
1. 广州中医药大学，广州 510006
- Keywords:
Herpesvirus 3, human;
Astrocytes;
Nod-like receptor protein 3;
Caspase 1;
Interleukin-1 beta
- From:
Chinese Journal of Dermatology
2024;57(6):524-529
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect of different concentrations and infection durations of varicella-zoster virus (VZV) on nucleotide-binding and oligomerization domain-like receptor protein 3 (NLRP3) inflammasome-related proteins in mouse brain astrocytes.Methods:Mouse brain astrocytes were infected with different dilutions (10 0 [undiluted original concentration] - 10 -7) of cell-free VZV suspension, and the cytopathic effect was observed under a microscope in each group to determine the concentration and infection duration of VZV for subsequent experiments. Some mouse brain astrocytes were infected with cell-free VZV suspension at dilutions of 10 0, 10 -2, 10 -4, and 10 -6, and samples were collected on day 5; some other mouse brain astrocytes were infected with cell-free VZV suspension at the median tissue culture infective dose (TCID 50) of 1.0 × 10 5/ml, and the samples were collected on days 3, 5, 7, and 9, separately; normally cultured mouse brain astrocytes served as control groups in the above experiments. Western blot analysis was performed to determine the protein expression of VZV glycoprotein E (gE), NLRP3, caspase-1, and interleukin-1β (IL-1β) in mouse brain astrocytes in each group. One-way analysis of variance was used for comparisons of data with a normal distribution, and least significant difference- t test was used for multiple comparisons. Results:On day 3 after infection with VZV at the dilution of 10 0, the number of cells decreased, some cells became irregular, lost their original shapes, and showed granular cytoplasm and an unclear nucleus structure, while some cells fused and formed multinucleated giant cells; on day 5, ≥ 50% of cells showed the aforementioned changes and some intercellular spaces were dilated giving a network-like appearance in the 10 0-dilution group; on day 9, the 10 0- - 10 -6- dilution groups all showed dilated intercellular spaces arranged in networks and cytopathic effect in ≥ 50% of cells; on day 10, some cells in the control group exhibited cytoplasmic vacuolation, exfoliation and flotation. Based on these results, the following experiments were conducted with VZV at a concentration gradient ranging from 10 0 to 10 -6 dilutions, and the observations were performed from day 3 to day 9. Western blot analysis showed increased protein expression of VZV gE, NLRP3, caspase-1, and IL-1β in the VZV infection groups compared with the control group, and there were significant differences among different VZV concentration groups ( F = 21.45, 43.82, 21.85, 29.53, respectively, all P < 0.05) and different infection duration groups ( F = 224.10, 78.79, 186.50, 85.08, respectively, all P < 0.05). With the increasing dilution ratios, the expression of inflammasome-related proteins (NLRP3, caspase-1, and IL-1β) decreased, with the highest expression observed in the 10 0-dilution group and the lowest in the 10 -6-dilution group (all P < 0.05). On days 3, 5, and 7 after VZV infection, the expression of NLRP3 inflammasome-related proteins increased over time, with the highest expression observed on day 7 and the lowest on day 3 (all P < 0.05), while their expression decreased on day 9. Conclusion:The expression of NLRP3 inflammasome-related proteins was up-regulated in mouse brain astrocytes after VZV infection, and their expression levels increased along with the increase in the VZV concentration and infection duration in a certain range.
