[关键词]
[摘要]
该研究探究了食用蘑菇杨树菇凝集素AAL(Agrocybe aegerita lectin)对小鼠肝损伤的作用机制。通过小鼠尾静脉注射AAL的方式,根据不同的处理时间分0 h、6 h和9 h三个实验组,分别检测其血清AST(天门冬氨酸氨基转移酶)和ALT(丙氨酸氨基转移酶),对肝脏组织切片进行HE染色,RT-PCR检测肝脏细胞因子表达,流式细胞术对肝脏细胞进行分析。与0 h组相比,对小鼠尾静脉注射AAL 6 h和9 h后,小鼠血清ALT和AST显著升高(p<0.001);肝组织切片HE染色发现小鼠肝脏呈现明显的肝损伤,可见坏死区和免疫细胞浸润。注射AAL 6 h后肝脏细胞因子IL-10、IL-33、IL-27、IFN-γ和TNF-α的mRNA表达分别升高到25.14倍、4.62倍、5.14倍、16.51倍和28.10倍。6 h组小鼠肝脏中的T细胞、NKT细胞、CD8+T细胞比例分别由27.92%显著升高到45.95%(p<0.05)、2.46%显著升高到6.36%(p<0.05)、17%显著升高到27.15%(p<0.05)。活化的CD69+T细胞和CD69+CD8+T细胞的比例分别由10.02%显著升高到18.28%(p<0.01)和5.96%显著升高到16.20%(p<0.01)。结果表明食用菌杨树菇AAL可诱导小鼠肝损伤,活化肝脏CD8+T细胞,显著上调细胞因子IL-10、IL-33、IL-27、IFN-γ和TNF-α的表达,提示食用菌凝集素成分是需要考虑安全使用剂量的。
[Key word]
[Abstract]
The mechanism of the lectin in edible mushroom Agrocybe aegerita (Agrocybe aegerita lectin, AAL) underlying its action on the liver injury in C57BL/6 mice was investigated. C57BL/6 mice were injected by AAL through the tail vein. According to different times of treatment, the mice were divided into three groups: 0 h, 6 h and 9 h, and the serum ALT (Alanine aminotransferase ) and AST (Aspartate aminotransferase) were measured, and the liver tissue sections were stained with HE, and the expression of liver cytokines were detected by RT-PCR, and he liver cells were analyzed by flow cytometry. At 6 h and 9 h groups, the 6 h and 9 h injection of AAL into the tail vein of mice significantly increased the serum ALT and AST (p<0.001); HE staining of liver tissue sections showed obvious liver damage, with necrotic areas and lymphocyte infiltration. Compared with the 0 h group, the serum ALT and AST of mice were significantly increased (p<0.001); the mRNA expression of cytokines IL-10, IL-33, IL-27, IFN-γ and TNF-α in the liver of the 6 h mouse group significantly increased by 25.14 times, 4.62 times, 5.14 times, 16.51 times and 28.10 times, respectively. The proportions of T cells, NKT cells, and CD8+T cells in the liver of the 6 h group significantly increased from 27.92% to 45.95% (p<0.05), 2.46% to 6.36% (p<0.05), and 17% to 27.15% (p<0.05), respectively. The proportions of activated CD69+T cells and CD69+CD8+T cells in the liver of the 6 h group significantly increased from 10.02% to 18.28% (p<0.01), and 5.96% to 16.20% (p<0.01), respectively. These results showed that AAL induced liver injury in mice, activated CD8+T cells, and significantly upregulated the mRNA expression of the liver cytokines IL-10, IL-33, IL-27, IFN-γ and TNF-α, suggesting that there is a need to consider the dose of the fungal lectin for safe use.
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[基金项目]
国家自然科学基金项目(81102850)