Fig. 4: Autophagy in BMDMs challenged with LPS was aggravated by CRH. | Cell Death Discovery

Fig. 4: Autophagy in BMDMs challenged with LPS was aggravated by CRH.

From: Corticotropin releasing hormone promotes inflammatory bowel disease via inducing intestinal macrophage autophagy

Fig. 4

BMDMs from C57BL/6 mice were isolated and treated with/without LPS (100 ng/mL)/CRH (10−8 M) for 24 h. IL-1β, IL-18, and TNF-α mRNA levels were analyzed by real-time qPCR. The levels of autophagy-related proteins were analyzed in western blots. Autophagosome number was evaluated using transmission electron microscopy. Autophagy flux was assessed using the mRFP-GFP-LC3 plasmid. A In comparison with LPS-treated BMDMs, BMDMs treated with LPS + CRH exhibited a significant increase in the mRNA levels of proinflammatory cytokines (n = 6 per group). **P < 0.01 vs. the control group; ##P < 0.01 vs. the LPS + vehicle group. B, C In comparison with the LPS + vehicle group, BMDMs in the LPS + CRH group demonstrated an increase in Beclin-1, an increase in the LC3-II/I ratio, and a decrease in p62/SQSTM1 (n = 5 per group). **P < 0.01 vs. the control group; ##P < 0.01 vs. the LPS + vehicle group. An increase in the number of autophagosomes (D, E) (n = 5 per group) and an increase in autophagy flux (F, G) were also observed (n = 6 per group). **P < 0.01 vs. the control group; ##P < 0.01 vs. the LPS + vehicle group.

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