Inhibition of PI3K/Akt/mTOR pathway by ammonium chloride induced apoptosis and autophagy in MAC-T cell. (May 2021)

• Record Type: Journal Article
• Authors: Feng, Luping; Liao, Hang; Liu, Jingsong; Xu, Chunmei; Zhong, Kai; Zhu, Heshui; Guo, Shuang; Guo, Yujie; Han, Liqiang; Li, Heping; Wang, Yueying
• Is Part Of: Research in veterinary science. Volume 136(2021)
• Page Start: 622
• Page End: 630
• Publication Date: 2021-05
• Subjects: NH3 -- MAC-T cells -- Apoptosis -- Autophagy -- PI3K/Akt/mTOR pathway; Veterinary medicine -- Periodicals; Veterinary Medicine -- Periodicals; Médecine vétérinaire -- Périodiques; Médecine vétérinaire -- Recherche -- Périodiques; Diergeneeskunde; 636.089
• DOI: 10.1016/j.rvsc.2021.01.020
• Languages: English
• ISSNs: 0034-5288

Abstract

Abstract: Ammonia is a harmful gas with a pungent odor, participates in the regulation of a variety of apoptosis and autophagy, which in turn affects the growth and differentiation of cells. To test the regulation of NH3 on the apoptosis and autophagy of mammary epithelial cells, we selected NH4 Cl as NH3 donor in vitro model. MTT and CCK-8 assay kits were employed to detect cell activity. Real-time quantitative PCR and western blot methods were used to detect the abundance of inflammatory molecules, apoptosis markers, and autophagy genes. We selected TUNEL kit and the Annexin-FITC/PI method to detect apoptosis. TEM analysis was used to detect autophagic vesicles, and MDC stain evaluated the formation of autophagosome. The results indicated that NH4 Cl reduced cell viability in a concentration-dependent manner and promoted cell inflammatory response, apoptosis, and autophagy. NH4 Cl stimulation notable increased the autophagosomes number. Interestingly, we also detected that the addition of LY294002 and Rapamycin inhibited the PI3K/Akt pathway and the mTOR pathway, respectively, resulting in changes in both apoptosis and autophagy. Therefore, we draw a conclusion that NH3 may regulate the apoptosis and autophagic response of bovine mammary epithelial cells through the PI3K/Akt/mTOR signaling pathway. Further investigations on ammonia's function in other physiological respects, will be critical to provide theoretical help for the improvement of production performance. It will be also helpful for controlling the harmful gas ammonia concentration in the livestock house to protect the health of dairy cows. Highlights: NH4 Cl inhibited the viability of MAC-T cell, depending on concentration and treatment time, and displaying dose-dependent effect. NH4 Cl aggravated the inflammation, apoptosis and autophagy in MAC-T cells through the PI3K/Akt/mTOR signaling pathway. Ammonia upregulated the apoptosis and autophagy of the cells. NH4 Cl stimulation notable increased the autophagosomes number. The PI3K/Akt/mTOR signals mediated the autophagy and apoptosis of MAC-T cells induced by NH4 Cl.