谷氨酰胺对BCG诱导小鼠传代巨噬细胞凋亡的调控作用

doi:10.11843/j.issn.0366-6964.2023.07.036

Abstract

Abstract: This experiment was to investigate the role of glutamine in the regulation of apoptosis in Bacille Calmette-Guerin vaccine (BCG)-infected macrophages. In this study, a cellular model of apoptosis caused by BCG infection was established using mouse mononuclear macrophages RAW264.7, and the optimal time of BCG infection and the number of infection replicates were clarified. Then, a glutamine deprivation model was constructed by glutamine-free medium and combined with BCG infection, and the Key metabolic markers of glutamine were detected by ELISA, immunoblotting, immunofluorescence and flow cytometry. The results showed that BCG infection significantly upregulated the expression of Caspase 3 and PARP in mouse macrophages RAW264.7 (P<0.001) and promoted the catabolism of glutamine. Glutamine deprivation significantly reduced glutamate (P<0.001) and glutathione (P<0.01) levels in BCG-infected macrophages and increased intracellular ROS content (P<0.001). Meanwhile, glutamine deprivation highly significantly up-regulated the expression of Caspase 3, PARP, Cytc and Bax (P<0.001) and highly significantly down-regulated the expression of the apoptosis suppressor protein Bcl-2 (P<0.001). These results indicated that glutamine deprivation in BCG-infected RAW264.7 cells promotes apoptosis by reducing intracellular glutathione levels, causing ROS accumulation and thus promoting apoptosis through an endogenous pathway.

Key words: BCG, glutamine, macrophage RAW264.7, apoptosis

CLC Number:

S855.2

AN Qi, YU Jialin, WU Xiaoling, DENG Guangcun. Regulation of BCG-induced Apoptosis in Murine Macrophages by Glutamine[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(7): 3054-3063.

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Regulation of BCG-induced Apoptosis in Murine Macrophages by Glutamine

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