RESUMO
Leucine, a kind of branched-chain amino acid, plays a regulatory role in the milk production of mammalian mammary glands, but its regulatory functions and underlying molecular mechanisms remain unknown. This work showed that a leucine-enriched mixture (LEUem) supplementation increased the levels of milk protein and milk fat synthesis in primary bovine mammary epithelial cells (BMECs). RNA-seq of leucine-treated BMECs indicated alterations in lipid metabolism, translation, ribosomal structure and biogenesis, and inflammatory response signaling pathways. Meanwhile, the supplementation of leucine resulted in mTOR activation and increased the expression of BCKDHA, FASN, ACC, and SCD1. Interestingly, the expression of PPARα was independently correlated with the leucine-supplemented dose. PPARα activated by WY-14643 caused significant suppression of lipogenic genes expression. Furthermore, WY-14643 attenuated leucine-induced ß-casein synthesis and enhanced the level of BCKDHA expression. Moreover, promoter analysis revealed a peroxisome-proliferator-response element (PPRE) site in the bovine BCKDHA promoter, and WY-14643 promoted the recruitment of PPARα onto the BCKDHA promoter. Together, the present data indicate that leucine promotes the synthesis of ß-casein and fatty acid and that PPARα-involved leucine catabolism is the key target.
Assuntos
Caseínas , PPAR alfa , Bovinos , Animais , Caseínas/genética , Caseínas/metabolismo , Leucina/farmacologia , Leucina/metabolismo , PPAR alfa/genética , PPAR alfa/metabolismo , Glândulas Mamárias Animais/metabolismo , Ácidos Graxos/metabolismo , Células Epiteliais/metabolismo , Mamíferos/metabolismoRESUMO
In this paper, we introduce a novel method for the fabrication of self-assembly plasmonic metamaterials by exploiting fluid instabilities of optical thin films. Due to interplay between template reflow and spinodal dewetting, two metal nanoparticles of different sizes are generated on the top mesas of free-standing porous anodic aluminum oxide (AAO) template, which results in the apprearance of double resonant peaks in the extinction spectrum. These two resonant peaks possess refractive index resolution 3.27 × 10-4 and 2.53 × 10-4 RIU, respectively. This optical intensity modulation based plasmonic nanoplatform shows a dramatically surface sensing performance with outstanding detection capacity of biomolecules, because of the very small decay length of electric field at dual-modes. The detection ability for concanavalin A (Con A) demonstrats that the limit of detection of dual-modes reaches as small as 68 and 79 nM, respectively.