Aims: High-expressed miR-330-3p in gestational diabetes mellitus (GDM) patients was reported. However, the role and mechanism of miR-330-3p in GDM are rarely reported. In this research, we aim to investigate the effects of miR-330-3p on GDM. Methods: MiR-330-3p expression in the GDM patients' blood was determined by q-PCR. Blood glucose of blood samples was detected using blood glucose detection kits. Glucokinase (GCK) was confirmed to be a target gene of miR-330-3p by bioinformatics and luciferase analysis. Correlations between miR-330-3p with GCK and blood glucose were analyzed by Pearson correlation analysis. After INS-1 cells were treated with glucose and transfected with mimic, inhibitor or siGCK, GCK expression was detected by western blot, and q-PCR, enzyme-linked immunosorbent assays, cell counting kit-8 and Annexin-V/propidium iodide were conducted to examine the expression of insulin, cell viability and apoptosis. Results: MiR-330-3p was high-expressed in GDM patients' blood, while GCK was low-expressed. The miR-330-3p expression level positively correlated with blood glucoseand and it was highly expressed in glucosetreated INS-1 cells (11 and 22 mmol/L), while miR-330-3p expression negatively correlated with GCK expression. GCK expression was inhibited by miR-330-3p mimic and enhanced by the miR-330-3p inhibitor. MiR-330-3p mimic inhibited INS-1 cells' insulin expression, cell viability and induced apoptosis. Yet miR-330-3p inhibitor and siGCK exhibited opposite effects which miR-330-3p mimic and GCK played on INS-1 cells. In addition, siGCK reversed the effect of miR-330-3p inhibitor on INS-1 cells. Conclusion: Our findings proved that miR-330-3p targeting GCK lead to the dysfunction of INS-1 cells in GDM, and could become a therapeutic target for GDM treatment.

A novel metabolomic method based on gas chromatography-mass spectrometry was applied to investigate serum metabolites in response to dietary Gln supplementation in piglets. Sixteen, 21-d-old pigs were weaned and assigned randomly to 2 isonitrogenous diets: 1) Gln diet, which contained 1% L-Gln (as-fed basis), and 2) control diet, which contained L-Ala to make this diet isonitrogenous with the Gln diet. Serum samples were collected to characterize metabolites after a 30-d treatment. in addition, 4 liver samples per treatment were collected to examine enzyme activity and gene expression involved in metabolic regulation. Results indicated that 12 metabolites were altered (P < 0.05) by Gln treatment, including carbohydrates, AA, and fatty acids. A leave-one-out cross validation of random forest analysis indicated that Pro was most important among the 12 metabolites. Thus, these data demonstrate that the control and Gln-supplemented pigs differed (P < 0.05) in terms of metabolism of carbohydrates, Pro, Tyr, and glycerophospholipids. Principal component analysis yielded separate clusters of profiles between the Gln and control groups. Metabolic enzyme activities of Ala aminotransferase and hexokinase increased by 26.8% (P = 0.026) and 26.2% (P = 0.004) in the liver of Gln-supplemented pigs vs. control, respectively, whereas pyruvate kinase (PK) activity decreased by 29.1% (P = 0.001). The gene expression of PK in the liver decreased by 66.1% (P = 0.034) by Gln treatment for 30 d. No differences were observed for the mRNA abundance of mammalian target of rapamycin and PPARγ. On the basis of these data, Gln treatment affected carbohydrate, lipid, and AA metabolism in the whole body of the early weaned piglets. These findings provide insight into specific metabolic pathways and lay the groundwork for the complex metabolic alteration in response to dietary Gln supplementation of pigs.