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Arginine Relieves the Inflammatory Response and Enhances the Casein Expression in Bovine Mammary Epithelial Cells Induced by Lipopolysaccharide.

Wu T, Wang C, Ding L, Shen Y, Cui H, Wang M, Wang H - Mediators Inflamm. (2016)

Bottom Line: The results showed that arginine reduced the LPS-induced production like IL-1β, IL-6, TNF-α, and iNOS.Though the expression of NF-κB was attenuated and the mTOR signaling pathway was upregulated, arginine had no effect on TLR4 expression.In addition, our results show that the content of β-casein and the total casein were enhanced after arginine was supplemented in LPS-induced BMECs.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Metabolic Manipulation of Herbivorous Animal Nutrition, College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China.

ABSTRACT
As one of functional active amino acids, L-arginine holds a key position in immunity. However, the mechanism that arginine modulates cow mammary inflammatory response in ruminant is unclear. Therefore, this study was conducted to investigate the effects of L-arginine on inflammatory response and casein expression after challenging the bovine mammary epithelial cells (BMECs) with lipopolysaccharide (LPS). The cells were divided into four groups, stimulated with or without LPS (10 μg/mL) and treated with or without arginine (100 μg/mL) for 12 h. The concentration of proinflammatory cytokines, inducible nitric oxide synthase (iNOS), mammalian target of rapamycin (mTOR), and Toll-like receptor 4 (TLR4) signaling pathways as well as the casein was determined. The results showed that arginine reduced the LPS-induced production like IL-1β, IL-6, TNF-α, and iNOS. Though the expression of NF-κB was attenuated and the mTOR signaling pathway was upregulated, arginine had no effect on TLR4 expression. In addition, our results show that the content of β-casein and the total casein were enhanced after arginine was supplemented in LPS-induced BMECs. In conclusion, arginine could relieve the inflammatory reaction induced by LPS and enhance the concentration of β-casein and the total casein in bovine mammary epithelial cells.

No MeSH data available.


Related in: MedlinePlus

Effect of Arg on the mRNA expression and phosphorylation levels of PI3K/AKT/mTOR signaling pathway in LPS-treated BMECs. Pure 2nd generation BMECs were starved for 16 h and then cultured for 12 h in DMEM/F12 medium containing 0 or 10 μg/mL LPS and 0 or 100 μg/mL Arg. The mRNA levels of PI3K, AKT, and mTOR were analyzed using qPCR. Protein expression and its phosphorylation level were analyzed using western bolt. β-Actin was used as an internal reference. Data shown are mean ± SEM of three independent experiments. Means with different letters (A, B, or C) are significantly different (P < 0.05) from each other.
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fig4: Effect of Arg on the mRNA expression and phosphorylation levels of PI3K/AKT/mTOR signaling pathway in LPS-treated BMECs. Pure 2nd generation BMECs were starved for 16 h and then cultured for 12 h in DMEM/F12 medium containing 0 or 10 μg/mL LPS and 0 or 100 μg/mL Arg. The mRNA levels of PI3K, AKT, and mTOR were analyzed using qPCR. Protein expression and its phosphorylation level were analyzed using western bolt. β-Actin was used as an internal reference. Data shown are mean ± SEM of three independent experiments. Means with different letters (A, B, or C) are significantly different (P < 0.05) from each other.

Mentions: The results of the relative gene expression of PI3K, AKT, and mTOR are shown in Figure 4. The results showed that the abundance of PI3K and mTOR gene expression in the LPS quarter was markedly lower (P < 0.05) than that in the control quarter, yet the abundance of AKT was not significantly different (P > 0.05). Compared with LPS quarter, the mRNA levels of PI3K, mTOR, and AKT in quarter treated with both LPS and arginine were markedly enhanced (P < 0.05). We also found that LPS dramatically decreased (P < 0.01) the level of p-mTOR/mTOR, but it had no effect on level of p-PI3K/PI3K and p-AKT/AKT (P > 0.05). In contrast to LPS quarter, the level of p-AKT/AKT and p-mTOR/mTOR in cells treated with LPS and arginine was notably augmented (P < 0.01), but the level of p-PI3K/PI3K was not affected (P > 0.05).


Arginine Relieves the Inflammatory Response and Enhances the Casein Expression in Bovine Mammary Epithelial Cells Induced by Lipopolysaccharide.

Wu T, Wang C, Ding L, Shen Y, Cui H, Wang M, Wang H - Mediators Inflamm. (2016)

Effect of Arg on the mRNA expression and phosphorylation levels of PI3K/AKT/mTOR signaling pathway in LPS-treated BMECs. Pure 2nd generation BMECs were starved for 16 h and then cultured for 12 h in DMEM/F12 medium containing 0 or 10 μg/mL LPS and 0 or 100 μg/mL Arg. The mRNA levels of PI3K, AKT, and mTOR were analyzed using qPCR. Protein expression and its phosphorylation level were analyzed using western bolt. β-Actin was used as an internal reference. Data shown are mean ± SEM of three independent experiments. Means with different letters (A, B, or C) are significantly different (P < 0.05) from each other.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig4: Effect of Arg on the mRNA expression and phosphorylation levels of PI3K/AKT/mTOR signaling pathway in LPS-treated BMECs. Pure 2nd generation BMECs were starved for 16 h and then cultured for 12 h in DMEM/F12 medium containing 0 or 10 μg/mL LPS and 0 or 100 μg/mL Arg. The mRNA levels of PI3K, AKT, and mTOR were analyzed using qPCR. Protein expression and its phosphorylation level were analyzed using western bolt. β-Actin was used as an internal reference. Data shown are mean ± SEM of three independent experiments. Means with different letters (A, B, or C) are significantly different (P < 0.05) from each other.
Mentions: The results of the relative gene expression of PI3K, AKT, and mTOR are shown in Figure 4. The results showed that the abundance of PI3K and mTOR gene expression in the LPS quarter was markedly lower (P < 0.05) than that in the control quarter, yet the abundance of AKT was not significantly different (P > 0.05). Compared with LPS quarter, the mRNA levels of PI3K, mTOR, and AKT in quarter treated with both LPS and arginine were markedly enhanced (P < 0.05). We also found that LPS dramatically decreased (P < 0.01) the level of p-mTOR/mTOR, but it had no effect on level of p-PI3K/PI3K and p-AKT/AKT (P > 0.05). In contrast to LPS quarter, the level of p-AKT/AKT and p-mTOR/mTOR in cells treated with LPS and arginine was notably augmented (P < 0.01), but the level of p-PI3K/PI3K was not affected (P > 0.05).

Bottom Line: The results showed that arginine reduced the LPS-induced production like IL-1β, IL-6, TNF-α, and iNOS.Though the expression of NF-κB was attenuated and the mTOR signaling pathway was upregulated, arginine had no effect on TLR4 expression.In addition, our results show that the content of β-casein and the total casein were enhanced after arginine was supplemented in LPS-induced BMECs.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Metabolic Manipulation of Herbivorous Animal Nutrition, College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China.

ABSTRACT
As one of functional active amino acids, L-arginine holds a key position in immunity. However, the mechanism that arginine modulates cow mammary inflammatory response in ruminant is unclear. Therefore, this study was conducted to investigate the effects of L-arginine on inflammatory response and casein expression after challenging the bovine mammary epithelial cells (BMECs) with lipopolysaccharide (LPS). The cells were divided into four groups, stimulated with or without LPS (10 μg/mL) and treated with or without arginine (100 μg/mL) for 12 h. The concentration of proinflammatory cytokines, inducible nitric oxide synthase (iNOS), mammalian target of rapamycin (mTOR), and Toll-like receptor 4 (TLR4) signaling pathways as well as the casein was determined. The results showed that arginine reduced the LPS-induced production like IL-1β, IL-6, TNF-α, and iNOS. Though the expression of NF-κB was attenuated and the mTOR signaling pathway was upregulated, arginine had no effect on TLR4 expression. In addition, our results show that the content of β-casein and the total casein were enhanced after arginine was supplemented in LPS-induced BMECs. In conclusion, arginine could relieve the inflammatory reaction induced by LPS and enhance the concentration of β-casein and the total casein in bovine mammary epithelial cells.

No MeSH data available.


Related in: MedlinePlus