CircN4bp1 Facilitates Sepsis-Induced Acute Respiratory Distress Syndrome through Mediating Macrophage Polarization via the miR-138-5p/EZH2 Axis

Figure 7

Knock down of circN4bp1 in macrophage alleviated lung injury induced by sepsis after CLP surgery through inhibition of M1 macrophage activation. (a) CLP surgery was performed on the vector () and circN4bp1-KD group () mice, and survival was monitored for 10 days. Mice were intravenously injected with plasmid transfected macrophages through the tail vein 24 hours before CLP. Treatment with si-circN4bp1 significantly improved long-term survival compared to the vector vehicle group. , log rank test for survival study. (b) qRT-PCR of quantification of the circN4bp1expression in the macrophages isolated from BALF of the four group mice (sham, sham+circN4bp1-KD, CLP+ vector, and CLP+ circN4bp1-KD group). (c) Lungs were fixed, sectioned, and stained with H&E. Representative sections are shown for the four above mentioned groups (original magnification, ×200). (d) Evaluation of wet/dry weight ratio is compared among the four abovementioned groups. (e) Quantification of protein levels in the bronchial alveolar lavage (BAL) in the four above mentioned groups. (f) Quantification of IL-6, TNF-α, and IL-10 levels in the BALF macrophages in the four abovementioned groups. (g) qRT-PCR quantification of miR-138-5p in the BALF macrophages in the four abovementioned groups. (h) Macrophages isolated from BALF were subjected to immunoblot analysis with specific antibodies against INOS, Arg-1, EZH2, p-STAT1, PPAR-γ, and GAPDH. Expression levels of indicated proteins were quantified by densitometry and normalized with GAPDH. All data are expressed as . (, vs. sham and Sham+circN4bp1-KD group, # vs. CLP+ vector group, determined by one-way ANOVA for multiple group comparisons).

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