Growth Hormone Receptor Signaling Inhibits Hepatic H2S Production In Vitro
(A–C) Endogenous H2S production in primary mouse hepatocytes as measured via two-photon fluorescent microscopy under different medium conditions: (A) +/−growth serum and +/−GH, with the asterisk indicating the significance of the difference between Complete and −Serum, and the # sign indicating the significance of the difference between −Serum and −Serum+GH, ∗/#p < 0.05; (B) +/−growth serum, +/−GH, +/−AZD1480, with the asterisk indicating the significance of the difference between Complete and −Serum in each group, and the # sign indicating the significance of the difference between −Serum (−) (no addition) and −Serum+ZD1480; and the $ sign indicating the significance of the difference between +GH and +GH+ZD1480 for both Complete and −Serum conditions, ∗/#/$p < 0.05; (C) +/−FGF21 in Complete medium containing serum, with the asterisk indicating the significance of the difference between Complete and Complete+FGF21; ∗p < 0.05.
(D) mRNA expression of Igf-I, Cgl, and Cbs in mouse primary hepatocytes. The asterisk indicates the significance of the difference between +GH and +GH+ZD1480; ∗p < 0.05.
(E and F) Endogenous H2S production in mouse primary hepatocytes by florescent microscopy after overnight treatment under the indicated medium conditions followed by addition of the P3 probe for 1 hr. The asterisk indicates the significance of the difference between Complete and −Serum, and the # sign indicates the significance of the difference between −Serum and −Serum+BAF (E), or −Serum−SAA and −Serum−SAA+CQ (F); ∗#p < 0.05.
(G and H) Endogenous H2S production in Hepa1-6 cells with or without knockdown of autophagy components ATG5 and ATG7 by shRNA (G) or in MEFs by genetic knockout of ATG5 and ATG7 (H) detected by UV spectrophotometry after overnight treatment with or without serum followed by addition of the P3 probe for 1 hr. The asterisk indicates the significance of the difference between Complete and −Serum in the Control group, and the # sign indicates the significance of the difference between −Serum Control and −Serum ATG5 deficient or −Serum ATG7 deficient; ∗/#p < 0.05. Each experiment was repeated at least three times. Scale bars, 100 μm (A–C, E, and F). Error bars are ± SEM. See also .