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

Figure 2

KL treatment improves cell viability, alleviates inflammatory response, and reduces apoptosis in PQ-exposed A549 cells. (a) A549 cell was preincubated with gradient concentrations of KL (50-400 pM) for 1 h then treated with or without 200 μM of PQ for 24 h. Cell viability was measured using CCK-8 assay. (b) Fluorescence image of Calcein-AM/PI costained A549 cells receiving corresponding treatments. The levels of the proinflammatory cytokines IL-1β (c) and IL-6 (d) in the culture supernatants were determined by ELISA. (e) Apoptosis ratio of A549 cells receiving different treatments. Apoptosis was detected using flow cytometry, followed by Annexin V-FITC/PI double staining. (f) Comparisons of apoptosis ratio of A549 cells. The apoptosis ratio was calculated including early and late apoptosis. (g) The activity of caspase-3 in cell lysates was tested by a caspase-3 activity kit. (h) JC-1 staining monitors the changes in ΔΨm, and the ratios of green/red fluorescence intensity indicate the mitochondrial depolarization. Data are shown as the . versus control group; ^## versus PQ group, ANOVA.

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