STAT6介导的巨噬细胞极化对布鲁氏菌胞内存活的影响

doi:10.11843/j.issn.0366-6964.2022.01.026

Abstract

Abstract: This study aimed to investigate the effect of STAT6 mediated macrophage polarization on intracellular survival of Brucella. Brucella smooth strain S2308(S2308) and rough vaccine strain RB51(RB51) were used to infect macrophages.M1-type macrophage marker factors p65, NOS2 and IL-1β were detected by qRT-PCR. mRNA expression levels of M2-type macrophage marker factors STAT6, ARG1 and IL-10 were detected by qRT-PCR. The expression of M1-type marker CD86 and M2-type marker CD206 was detected by flow cytometry. The inhibitory effect of p-STAT6 protein and inhibitor AS on the protein was detected by Western Blot.The expression levels of M1-type cytokines TNF-α and IL-12 and M2-type cytokines IL-4 and IL-10 were detected by ELISA. Finally, CFU count of intracellular colonies was carried out. The qRT-PCR results showed that the mRNA expression of M1 type factor was significantly induced at 8 h and 12 h after infection, the expression of M2 type factor was low at 72 h, and the expression of M2 type factor was high at 72 h. Flow cytometry showed that S2308 could significantly induce CD86 expression at 12 h after infection, and CD206 expression at 72 h after infection, but RB51 had no effect on both.Western Blot results showed that strain S2308 activated STAT6 signaling pathway 72 h after infection, while RB51 hardly activated STAT6 signaling pathway. The inhibitory effect of inhibitor AS was the best at 2 μmol·L^-1 concentration. ELISA results showed that AS inhibitors could significantly inhibit the release of IL-4 and IL-10, and promote the release of TNF-α and IL-12.CFU count results showed that intracellular bacteria in S2308 group were firstly decreased and then significantly increased, and S2308+IL-4 group was significantly higher than S2308 group, S2308+AS group was significantly lower than S2308 group, and there was no significant difference in RB51 group. Brucella S2308 can induce the transformation of M1 type macrophages to M2 type macrophages through STAT6 in the late stage of infection, and promote the release of Th2 type cytokines, which is conducive to the intracellular survival of Brucella. RB51 does not activate this pathway and does not affect intracellular survival.

Key words: Brucella, macrophage polarization, STAT6 signal path, intracellular survival

CLC Number:

S852.614

XI Jing, WANG Yueli, DENG Xiaoyu, YANG Qin, LI Peidong, ZHANG Jiangwei, SUN Tianhao, ZHU Liangquan, YI Jihai, CHEN Chuangfu. Effect of STAT6 Mediated Macrophage Polarization on Intracellular Survival of Brucella[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(1): 263-271.

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Effect of STAT6 Mediated Macrophage Polarization on Intracellular Survival of Brucella

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