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目的:观察小鼠巨细胞病毒(MCMV)播散性感染急性期脾组织病理损伤程度与病毒量、caspase-1和下游促炎因子IL-1β、IL-18表达的关系,探讨MCMV感染后脾脏的免疫病理损伤机制。方法:建立MCMV全身播散性感染模型,MCMV Sminth株接种后第3、7、14和第28天各处死3只小鼠;同时设模拟感染小鼠作为对照。标准空斑试验检测脾组织病毒滴度;Western blot法检测脾脏中caspase-1表达强度;免疫组化法检测脾组织中IL-1β和IL-18表达状况,HE染色后用半定量法评估脾脏组织病理损伤程度;分析病毒滴度及caspase-1、IL-1β和IL-18表达与组织病理损伤的关系。结果:MCMV感染后,脾组织病毒滴度在感染后3 d升高,7 d明显下降,14 d时用标准空斑试验已检测不到病毒;与模拟感染对照组比较,MCMV感染组感染后第3天脾组织中caspase-1表达明显升高后逐渐下降(P<0.01);脾组织中IL-1β和IL-18表达呈进行性升高,于感染后7 d达峰值,14 d减少,28 d基本接近正常;脾组织病理损伤呈进行性加重,至感染后14 d达高峰,28 d明显减轻;IL-1β和IL-18在脾组织中的高表达早于其组织病理损伤,随着IL-1β和IL-18表达水平的下降,病理损害也逐渐恢复。结论:巨细胞病毒感染引起caspase-1表达增加,促进炎性因子(IL-1β、IL-18)合成和释放增加。IL-1β、IL-18不仅有抗病毒作用,同时也可能参与MC-MV播散性感染后脾组织的免疫病理损伤。
Objective: To observe the relationship between the degree of pathological damage and the expression of viral load, caspase-1 and the downstream pro-inflammatory cytokines IL-1β, IL-18 in mice infected with cytomegalovirus (MCMV) Immune pathological damage mechanism. Methods: The model of systemic disseminated infection of MCMV was established. Three mice were sacrificed on the 3rd, 7th, 14th and 28th days after inoculation of MCMV Sminth strain respectively. At the same time, mock-infected mice were used as the control. The spleen tissue virus titer was detected by standard plaque assay; the expression of caspase-1 in spleen was detected by Western blot; the expression of IL-1β and IL-18 in spleen was detected by immunohistochemical method; The degree of histopathological damage was analyzed. The relationship between the titer of virus and the expression of caspase-1, IL-1β and IL-18 and histopathological damage were analyzed. Results: After MCMV infection, the virus titer in spleen tissue increased 3 d after infection and decreased significantly at 7 d, and no virus was detected by standard plaque assay on the 14th day. Compared with the mock-infected control group, On day 3, the expression of caspase-1 in spleen tissue was significantly increased (P <0.01), while the expression of IL-1β and IL-18 in spleen increased progressively and reached its peak on the 7th day after infection and decreased on the 14th day , 28 d were almost normal; pathological injury of the spleen increased progressively and reached the peak on the 14th day after infection and was significantly reduced on the 28th day. The high expression of IL-1β and IL-18 in the spleen tissue was earlier than that of the histopathological injury, As the expression of IL-1β and IL-18 decreased, pathological changes gradually recovered. CONCLUSION: The cytomegalovirus infection causes caspase-1 expression and increases the synthesis and release of inflammatory cytokines (IL-1β, IL-18). IL-1β, IL-18 not only have antiviral activity, but also may be involved in immunopathological damage of splenic tissue after MC-MV disseminated infection.