Effect of different nitrogen sources on apoptosis of rumen epithelial cells
In this experiment,cell proliferation assay,flow cytometry assay,Western blot assay,Hochest33342 staining fluorescence microscopy assay were used to explore the effects of different nitrogen sources(methionine and ammonium chloride)on the apoptosis of rumen epithelial cells and their possible regulatory mechanisms.The results showed that for the rumen epithelial cells,the OD values of both the 0.12 mmol NH4Cl-added group and the 0 mmol Met-deficient group reached half of those of the control group at 36 h of treatment,which were significantly lower than those of the 0.048 mmol Met group,the 0.12 mmol Met group(control group)and the 0.048 mmol NH4Cl group(P＜0.05);the total apoptosis rate and late apoptosis rate were the highest in the 0 mmol Met group,which were significantly higher than those in the 0.12 mmol Met group(control group)and 0.12 mmol NH4Cl group(P＜0.05),while the early apoptosis rate in the 0 mmol Met group and 0.12 mmol NH4Cl group were significantly higher than those in the 0.12 mmol Met group(control group)(P＜0.05);the mitochondrial membrane potential was significantly decreased to 71.99％and 67.67％in the 0.12 mmol NH4Cl group and 0 mmol Met group,respectively,compared with the control group.The expression of the pro-regulatory proteins Caspase-3,Bax and p53 were significantly up-regulated in the 0 mmol Met group and 0.12 mmol NH4Cl group(P＜0.05)and the expression of the inhibitory proteins Bcl-2 and Bcl-xl were significantly down-regulated(P＜0.05)in the 0 mmol Met and 0.12 mmol NH4Cl groups compared with the control group;the expression of p-mTOR and p-AMPK proteins were significantly up-regulated(P＜0.05)in the 0 mmol Met and 0.12 mmol NH4Cl groups compared with the control group,while the expression of PI3K and p-AKT proteins were significantly down-regulated(P＜0.05).The above results indicated that methionine deficiency induced apoptosis in rumen epithelial cells,and NH4Cl substitution could alleviate the occurrence of apoptosis,and the induction process could be accomplished through PI3K-Akt-mTOR signaling pathway and AMPK-mTOR signaling pathway.This result illustrated the indispensable importance of methionine,which could provide reference significance for the study of compound feed development in ruminants.
万方数据
维普
