Mitogen-activated protein kinase-interacting kinase 1 is involved in mTOR-mediated pancreatic exocrine function of bovine pancreatic acinar cells with leucine supply.

Leucine improves the exocrine capacity of the cow pancreas, but the mechanism was not revealed clearly. Mitogen-activated protein kinase-interacting kinase 1 (MNK1) is a pancreatic acinar cell-specific stress response kinase that regulates digestive enzyme abundance. We aimed to investigate the MNK1 gene and protein expression profiles among various organs or tissues of dairy cows and to demonstrate the mechanism by which leucine-stimulated MNK1 regulates pancreatic exocrine function. Firstly, the expression profiles of MNK1 protein and gene in the tissues and organs of dairy cows were measured using immunohistochemistry and RT-qPCR methods. Next, an in vitro model of cultured Holstein dairy calf pancreatic acinar cells was used to detect the role of MNK1 during pancreatic enzymes release which is stimulated by leucine. Cells were incubated in culture medium containing L-leucine (0.45 mM) for 180 min, and samples were collected hourly, with the control not containing L-leucine (0 mM). MNK1 was expressed at very high levels in the pancreatic tissue of dairy cows. Leucine supplementation increased the α-amylase level but not lipase level at three time-points (60, 120, and 180 min), and the interaction between treatments and times was significant only for α-amylase. Leucine treatment enhanced (P < 0.05) the phosphorylation of MNK1 and eIF4E. In addition, inhibition of MNK1 decreased leucine-mediated α-amylase and lipase release (P < 0.05) and the phosphorylation of Mnk1 and eIF4E but did not affect (P > 0.05) the phosphorylation of the mTOR signalling pathway factors 4EBP1 and S6K1. In summary, MNK1 is a key regulator of pancreatic exocrine function, which is regulated by leucine in the pancreas of dairy cows.

Rapid Identification of unstable acyl glucoside flavonoids of Oxytropis racemosa Turcz by high-performance liquid chromatography-diode array detection-electrospray ionisation/multi-stage mass spectrometry.

INTRODUCTION: Oxytropis racemosa Turcz is an important minority medicine that is used mainly to improve children's indigestion, especially in inner Mongolia and Tibet. Previous studies indicated that the characteristic constituents of this plant are acylated flavonoids. OBJECTIVE: Rapidly identify the characteristic chemical constituents of O. racemosa by high-performance liquid chromatography-diode array detection-electrospray ionisation/multi-stage mass spectrometry (HPLC-DAD-ESI/MS(n) ) and suggest a useful method to control the quality of this medicinal plant. METHODS: In the HPLC fingerprint, 32 flavonoids were tentatively identified by a detailed analysis of their mass spectra, UV spectra and retention times. Furthermore, 13 flavonoids were confirmed by comparison with previously isolated compounds obtained from O. racemosa. RESULTS: In total, 32 flavonoids, including 13 flavonoids with 3-hydroxy-3-methylglutaric acid (HMG) moieties and four flavonoids with 3-malonyl moieties, were identified in the extract of O. racemosa. Among the compounds identified, 10 were characterised as new compounds for their particular acylated sugar moieties. CONCLUSIONS: The method described is effective for obtaining a comprehensive phytochemical profile of plants containing unstable acylated flavonoids. The method is also useful for constructing the chromatographic fingerprint of the minority medicine -O. racemosa Turcz for quality control.

Flavonol glycosides and monoterpenoids from Potentilla anserina.

Potentilin A (1), a rare diflavonol ester of mu-truxinic acid and a new normonoterpenoid, 2,6-dimethyl-2,3-dihydro-4-oxo-4H-pyran-2-acetic acid (2), was isolated from Potentilla anserina, together with 19 known flavonol glycosides (3-21) and 2 known monterpenoids (22,23). Their structures were elucidated by means of UV, IR, HR-ESI-MS, 1D and 2D NMR spectroscopic data.