Wei Sheng Yan Jiu. 2022 Jul;51(4):632-637. doi: 10.19813/j.cnki.weishengyanjiu.2022.04.022.
ABSTRACT
OBJECTIVE: To investigate the regulation mechanism of N-acetylcysteine(NAC) on cadmium-induced apoptosis of mouse testicular interstitial cells based on protein kinase B pathway(AKT pathway).
METHODS: Mouse testicular mesenchymal cells(TM3) were divided into fourgroups according to different treatment, control group, cadmium group(Cd, 5, 10, 20, 30, 40 and 50 μmol/L), NAC group(NAC, 500 μmol/L) and NAC+Cd group(500 μmol/L NAC+20 μmol/L Cd). Cells of NAC+Cd group were pretreated with NAC for 30 min, and then combined with cadmium for 24 h. Cell viability was determined by CCK8. Hoechst staining was used to determine cell morphology. Cell apoptosis rate was analyzed by flow cytometry. Malondialdehyde(MDA) and glutathione(GSH) were measured simultaneously. Western blot was used to detect the expression levels of AKT protein, B-cell lymphoma-2(Bcl-2) and Bcl-2 associated X protein(Bax).
RESULTS: Cadmium inhibited the proliferation of TM3 cells in a dose-effect relationship. Cell morphology observation showed that with the increase of cadmium concentration, the cells shrank, became round and even fell off, and appeared dense nuclear staining. The MDA level in Cd group was(1.56±0.11) μmol/mg prot, which was significantly higher than that in control group(P<0.01). Compared to the control group, the level of GSH was significantly decreased to(1.28±0.25) μmol/mg prot(P<0.01). NAC pretreatment could reduce the MDA content and increase the GSH level, and the difference was statistically significant compared with the Cd group(P<0.01). Western blot result showed that NAC pretreatment significantly increased levels of phosphorylated AKT and Bcl-2, the levels were 0.65±0.05 and 0.45±0.03, respectively(P<0.01). The Bax/Bcl-2 ratio was 1.54±0.15, which was significantly lower than that of the Cd group(P<0.01).
CONCLUSION: NAC can inhibit cadmium-mediated TM3 cell damage and apoptosis, which may be related to the improvement of oxidative stress state, activation of TM3 AKT pathway and reduction of Bax/Bcl-2 ratio.
PMID:36047270 | DOI:10.19813/j.cnki.weishengyanjiu.2022.04.022
