Figure 4.

(A) mRNA expression of p53/AMPK signaling pathway-related genes, which were detected via reverse transcription-quantitative polymerase chain reaction, indicated markedly higher expression levels in groups 2, 3, 4, 5 and 6 compared with group 1. (B-D) Western blotting was used to detect the expression levels of p53/AMPK signaling pathway-related proteins. The expression levels of mTOR, p-mTOR, p53, AMPKα and sestrin were increased in the experimental groups compared with group 1. Furthermore, when compared with group 5, p53, mTOR, p-mTOR, AMPKα and sestrin levels were significantly increased in groups 2, 3 and 4. Results for groups 2, 3 and 4 suggest that GZYKF activates mTOR, p-mTOR, AMPKα and sestrin, and inhibits p53 in a dose-dependent manner. Data are presented as mean ± standard error of the mean of three independent experiments. *P<0.05, **P<0.01, vs. group 5. p53, tumor suppressor p53; AMPK, adenosine monophosphate-activated protein kinase; TSC, tuberous sclerosis protein; mTOR, mechanistic target of rapamycin; p, phosphorylated; TP, testosterone propionate; GZYKF, Gui Zhu Yi Kun formula; group 1, control; group 2, treated with TP and high-dose GZYKF-containing serum; group 3, treated with TP and medium-dose GZYKF-containing serum; group 4, treated with TP and low-dose GZYKF-containing serum; group 5, treated with TP and serum without GZYKF; group 6, treated with TP.