Klotho Alleviates Lung Injury Caused by Paraquat via Suppressing ROS/P38 MAPK-Regulated Inflammatory Responses and Apoptosis

Figure 4

KL exerts protective roles on PQ-treated A549 cells by inhibiting ROS/P38 MAPK signaling. (a) The expression of P38 MAPK, JNK, and ERK was analyzed by western blotting. (b) Quantification of relative intensity of p-P38 MAPK, p-JNK, and p-ERK protein bands for each group. (c) Knockdown of P38 MAPK by siRNA was confirmed by western blot analysis. A549 cells were preincubated with P38-MAPK siRNA (50 nM) or SB203580 (10 μM), subsequently treated with or without 200 μM of PQ for 24 h. Cell viability (d) was measured using CCK-8 assay, and ROS production (e) was tested by DCFH-DA staining. The contents of IL-1β (f) and IL-6 (g) in the culture supernatants were evaluated by ELISA. The activity of caspase-3 (h) was detected by the caspase-3 activity kit, and the changes in ΔΨm (i) were detected by JC-1 staining.

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