Energy metabolism disorder mediated ammonia gas-induced autophagy via AMPK/mTOR/ULK1-Beclin1 pathway in chicken livers

Ammonia gas is a well-known environmental pollution gas, threatening human health. Ammonia gas is also one of the most harmful gases to livestock and poultry for many years. Many studies have demonstrated toxic effect of ammonia gas on animal health, such as eyes, respiratory system, and digestive system. However, the effect of ammonia gas toxicity on chicken livers and underlying molecular mechanism remains unclear. In this study, we selected chicken liver as research object and duplicated successfully ammonia gas poisoning model of chickens. 1-day-old Ross-308 broilers were randomly divided into the control group (the low ammonia gas group), and two treatment groups (the middle ammonia gas group and the high ammonia gas group) (3 replicates per group and 12 chickens per replicate). Ammonia gas concentration in the low ammonia gas group was <= 5 mg/m3 during day 1-42. Ammonia gas concentration in the middle group was set as 10 +/- 0.5 mg/m3 during day 1-21, and 15 +/- 0.5 mg/m3 during day 22-42). Ammonia gas concentration in the high ammonia gas group was set as 20 +/- 0.5 mg/ m3 during day 1-21, and 45 +/- 0.5 mg/m3 during day 22-42. The ultrastructure of chicken livers was observed. The activities of four ATPases (Na+K+-ATPase, Mg++-ATPase, Ca++-ATPase, and Ca++Mg++-ATPase), the expression of twelve energy metabolism-related genes (HK1, HK2, PK, PFK, PDHX, CS, LDHA, LDHB, SDHA, SDHB, avUCP, and AMPK), as well as the expression of ten autophagy-related genes (PI3K, LC3I, LC3II, Beclin1, SQSTM1, mTOR, ULK1, ATG5, ATG12, and ATG13) were measured to explore the effect of ammonia gas on energy metabolism and autophagy in chicken livers. Our results showed that excess ammonia gas induced mitochondrial and autophagic damage in chicken liver tissue cells. Meanwhile, ATPases activities were inhibited and the expression of energy metabolism-related genes changed during ammonia gas treatment, meaning that excess ammonia gas caused energy metabolism disorder. Furthermore, ammonia gas exposure altered the expression of autophagy-related genes, suggesting that ammonia gas treatment caused autophagy in chicken livers. Moreover, ammonia gas-induced AMPK compensatory up-regulation activated autophagy process through inhibiting mTOR and promoting ULK1. In addition. there were dose-dependent and time-dependent effects on all detected indexes in ammonia gas-caused chicken liver cell damage. Taken together, AMPK/mTOR/ULK1-Beclin1 pathway participated in energy metabolism disorder-mediated autophagic injury caused by ammonia gas exposure in chicken livers.

Automated summary

This study investigated the toxic effects of ammonia gas on chicken liver, revealing damage to mitochondrial and autophagy in chicken liver tissue cells, as well as disruption of energy metabolism. Exposure to excess ammonia gas induced autophagy in chicken livers, with the AMPK/mTOR/ULK1-Beclin1 pathway playing a role in this process. Dose-dependent and time-dependent effects were observed on all detected indexes in ammonia gas-induced chicken liver cell damage.