Rhodiola crenulata and Its Bioactive Components, Salidroside and Tyrosol, Reverse the Hypoxia-Induced Reduction of Plasma-Membrane-Associated Na,K-ATPase Expression via Inhibition of ROS-AMPK-PKCξ Pathway

Figure 3

RCE, salidroside, and tyrosol prevent hypoxia-mediated Na,K-ATPase endocytosis in A549 cells. The analysis of the distribution of α1-Na,K-ATPase and 𝛽-actin in the plasma membrane (PM) and post-plasma membrane (PPM) fractions by western blotting (a). The A549 cells were exposed to hypoxia (1% O₂) for 24 h, and the expressions of α1-Na,K-ATPase, PKCζ, p-AMPK, SOD2, and GPx2 were analyzed with or without a 30 min pretreatment with RCE (b), salidroside (c), and tyrosol (d) in the PM and PPM. Clathrin served as a loading control. (e)–(g) and (h)–(j) are the quantitative data for the relative levels of α1-Na,K-ATPase and PKCζ in the PM from (b), (c), and (d). (k)–(m) and (n)–(p) are the quantitative data for the relative levels of α1-Na,K-ATPase and p-AMPK in the PPM from (b), (c), and (d). The results represent the mean ± SEM of three independent experiments. ^∗𝑃<0.05; ^∗∗𝑃<0.01; ^∗∗∗𝑃<0.001 versus control; ^#𝑃<0.05; ^##𝑃<0.01; ^###𝑃<0.001 versus hypoxia (H).

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