ABSTRACT
MicroRNAs (miRNAs), a class of small non-coding RNAs, have emerged as novel and potent regulators of adipogenesis. However, few miRNAs have been fully investigated in porcine adipogenesis, given the fact that pig is not only an apropos model of human obesity research, but also a staple meat source of human diet. In this study, we showed that miRNA-199a-5p is highly expressed in porcine subcutaneous fat deposits compared to several other tissue types and organs measured alongside. Overexpression of miR-199a-5p in porcine preadipocytes significantly promoted cell proliferation while attenuating the lipid deposition in porcine adipocytes. By target gene prediction and experimental validation, we demonstrated that caveolin-1 (Cav-1) may be a bona fide target of miR-199a-5p in porcine adipocytes, accounting for some of miR-199a-5p's functions. Taken together, our data established a role of miR-199a-5p in porcine preadipocyte proliferation and differentiation, which is at least partially played by downregulating Cav-1.
Subject(s)
Adipocytes/cytology , MicroRNAs/metabolism , 3' Untranslated Regions , Adipocytes/metabolism , Adipogenesis , Animals , Base Sequence , Caveolin 1/chemistry , Caveolin 1/genetics , Caveolin 1/metabolism , Cell Proliferation , Cells, Cultured , Down-Regulation , Humans , MicroRNAs/chemistry , RNA, Messenger/metabolism , Sequence Alignment , SwineABSTRACT
MicroRNAs constitute a class of ~22-nucleotide non-coding RNAs. They modulate gene expression by associating with the 3' untranslated regions (3' UTRs) of messenger RNAs (mRNAs). Although multiple miRNAs are known to be regulated during myoblast differentiation, their individual roles in muscle development are still not fully understood. In this study, we showed that miR-199a-3p was highly expressed in skeletal muscle and was induced during C2C12 myoblasts differentiation. We also identified and confirmed several genes of the IGF-1/AKT/mTOR signal pathway, including IGF-1, mTOR, and RPS6KA6, as important cellular targets of miR-199a-3p in myoblasts. Overexpression of miR-199a-3p partially blocked C2C12 myoblast differentiation and the activation of AKT/mTOR signal pathway, while interference of miR-199a-3p by antisense oligonucleotides promoted C2C12 differentiation and myotube hypertrophy. Thus, our studies have established miR-199a-3p as a potential regulator of myogenesis through the suppression of IGF-1/AKT/mTOR signal pathway.
Subject(s)
Insulin-Like Growth Factor I/metabolism , MicroRNAs/metabolism , Proto-Oncogene Proteins c-akt/metabolism , TOR Serine-Threonine Kinases/metabolism , 3' Untranslated Regions , Animals , Base Sequence , Cell Line, Tumor , Humans , Mice , MicroRNAs/antagonists & inhibitors , Muscle, Skeletal/metabolism , Myoblasts/cytology , Myoblasts/metabolism , Oligonucleotides, Antisense/metabolism , Signal TransductionABSTRACT
OBJECTIVE: To establish chemical pattern recognition method for the identification and evaluation of Atractylodes macrocephala. METHOD: The chemical constituents in methanol extract of 32 samples of A. macrocephala were determined by HPLC. The fingerprints were obtained and were handled by hierarchical clustering analysis and stepwise discriminant analysis. RESULT AND CONCLUSION: According to the result of classification, all samples collected were devided into three Grades--the superior, the ordinary and the fake. Chemical pattern recognition method was established. It may be of practical value for the quality control of A. macrocephala.
