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. Author manuscript; available in PMC: 2021 Nov 16.
Published in final edited form as: Clin Sci (Lond). 2021 Jan 29;135(2):409–427. doi: 10.1042/CS20201340

Figure 6. Gastrin reduces Ang II-induced RPT cell apoptosis and enhances the efferocytosis of macrophages through PPAR-α.

Figure 6.

(A) RAW264.7 cells were incubated with saline vehicle (control, 25 μl) or gastrin (10−7 mol/l) for 24 h prior to measuring the mRNA expression of phagocytosis-associated genes. n=5; *P<0.05 vs. Vehicle treatment. (B) RAW264.7 cells were incubated with saline vehicle (control, 25 μl), CI988 (10−7 mol/l), gastrin (10−7 mol/l) or gastrin (10−7 mol/l) + CI988 (10−7 mol/l) for 24 h, prior to measuring PPAR-α mRNA. n=5; *P<0.05 vs. others; #P<0.05 vs. Gastrin treatment. (C) Primary cultures of peritoneal macrophages from WT and CCKBR−/− mice were incubated with saline vehicle (control, 25 μl) or gastrin (10−7 mol/l) for 24 h, prior to measuring PPAR-α mRNA. n=5; *P<0.05 vs. WT+Vehicle treatment; #P<0.05 vs. WT+Gastrin treatment. (D) RPT cells were incubated with saline vehicle (control, 25 μl), CI988 (10−7 mol/l), gastrin (10−7 mol/l), or gastrin (10−7 mol/l)+CI988 (10−7 mol/l) for 24 h, prior to measuring PPAR-α mRNA. n=5; *P<0.05 vs. others; #P<0.05 vs. Gastrin treatment. (E) Representative fluorescent images showing engulfment of apoptotic Jurkat cells (BCECF AM) by RAW264.7 cells (Dil) (left panel). RAW264.7 cells were infected with lentivirus expressing PPAR-α shRNA or control shRNA, and then treated with saline vehicle (control, 20 μl) or gastrin (10−7 mol/l) for 24 h prior to measuring efferocytosis (right graph). n=4; *P<0.05 vs. Vehicle treatment; #P<0.05 vs. Gastrin treatment. (F) RAW264.7 cells were infected with lenti-shPPAR-α or lentiviral particles vehicle (scramble shRNA transfected with lentiviral particles used as vehicle control) for 72 h, prior to gastrin (10−7 mol/l) treatment for 24 h. Efferocytosis-related mRNA was quantified by qRT-PCR. n=5; *P<0.05 vs. Vehicle treatment; #P<0.05 vs. Gastrin treatment. (G) Western blots of cleaved caspase 3 (marker of apoptosis) in RPT cells. RPT cells were infected with lenti-shPPAR-α or lentiviral particles vehicle (scramble shRNA transfected with lentiviral particles used as vehicle control), and then treated with Ang II (10−8 mol/l), alone or with gastrin (10−7 mol/l) for 24 h prior to Western blotting. n=5; *P<0.05 vs. Ang II treatment; #P<0.05 vs. Ang II+Gastrin treatment.