Mechanisms of macrophage phenotype polarization based on the TonEBP/NF-κB pathway under high salt intervention

ZHU Zhe; YANG Guohong; ZHAO Jihong

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Medical Journal of the Chinese People Armed Police Forces ›› 2021, Vol. 32 ›› Issue (8) : 676-680.

Mechanisms of macrophage phenotype polarization based on the TonEBP/NF-κB pathway under high salt intervention

ZHU Zhe¹, YANG Guohong², ZHAO Jihong²

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Abstract

Objective To investigate the possible mechanism of macrophage phenotypic polarization under hypersaline stimulation so as to provide data for future research into the prevention and treatment of salt-sensitive hypertension and other macrophage related cardiovascular diseases.Methods Using Raw264.7 mouse macrophages as the subjects, a TonEBP lentivirus interference stable cell line was constructed. The interference efficiency of TonEBP gene was detected by real-time PCR , and the cell lines of the infected group with the highest interference efficiency were selected for subsequent experiments.The mRNA expression levels of TonEBP、NF-κB 、M1-type and M2-type macrophage phenotypic markers in each group were detected by real-time PCR after twenty hours of corresponding intervention.Western blot was used to detect the protein expression levels of TonEBP、NF-κB、p-NF-κB and p-IKKα/β.Results Stable interfering cell lines of TonEBP-shRNA were constructed, and the TonEBP interfering efficiency was over 70% compared with the control group (P＜0.05).The mRNA expression levels of TonEBP、NF-κB andM1 macrophage phenotypic markers in the HS group were significantly up-regulated,(P＜0.05), while the mRNA expression levels of M2 type macrophage phenotypic markers were significantly down-regulated(P＜0.05).mRNA expression of M1-type macrophage phenotypic markers was significantly down-regulated,(P＜0.05) while mRNA expression of M2-type macrophage phenotypic markers was significantly up-regulated by interfering with the expressions of TonEBP and NF-κB respectively under high salt stimulation(P＜0.05).Compared with the control group, the protein expression levels of TonEBP、p-IKKα/β、NF-κB and p-NF-κB in the HS group were significantly up-regulated(P＜0.05), and the ratio of p-NF-κB to NF-κB was increased(P＜0.05).Compared with the HS group, protein expression levels of p-IKKα/β、NF-κB and NF-κB were significantly decreased after interference with the expression of TonEBP and NF-κB respectively under high salt stimulation(P＜0.05),but difference in the ratio of p-NF-κB to NF-κB was of no statistical significance(P＞0.05).Conclusions Activation of TonEBP/NF-κB signaling pathway is a possible mechanism of Raw264.7 cell shifting to M1 phenotype under high salt stimulation.The regulation of TonEBP/ NF-κB signaling pathway can change the direction in which the phenotypes of Raw264.7 cells are polarized under high salt induction.

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high salt / cardiovascular disease / macrophages / phenotype / signaling pathway

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ZHU Zhe, YANG Guohong, ZHAO Jihong. Mechanisms of macrophage phenotype polarization based on the TonEBP/NF-κB pathway under high salt intervention[J]. Medical Journal of the Chinese People Armed Police Forces. 2021, 32(8): 676-680

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