Journal: Nutrients

Article Title: Mulberry Leaf ( Morus alba L.) Extracts and Its Chlorogenic Acid Isomer Component Improve Glucolipotoxicity-Induced Hepatic Lipid Accumulation via Downregulating miR-34a and Decreased Inflammation

doi: 10.3390/nu14224808

Figure Lengend Snippet: Effects of MLE and nCGA on multiple signaling pathways in OH−induced HepG2 cells. HepG2 cells were co-treated with OH, MLE (3 mg/mL), and nCGA (50 or 150 μM) at 37 °C for 24 h. Protein extracts were measured by Western Immunoblotting; α-tubulin was used as an internal control. MLE and nCGA activated ( A ) pAMPK/AMPK and ( B ) PPARα/CPT-1 signaling pathways in OH-induced HepG2 cells, but reduced ( C ) SREBP-1/FASN and ( D ) SREBP-2/HMG-CoAR protein expressions. Values are the mean ± SD of three independent experiments. # p < 0.05 vs. the control group. * p < 0.05 vs. the OH group.

Article Snippet: Monoclonal antibodies against phosphor-AMPK (#2535), FASN (#3180) and α-tubulin were obtained from Cell Signaling Technology Inc (Beverly, MA, USA).

Techniques: Western Blot

Journal: Nutrients

Article Title: Mulberry Leaf ( Morus alba L.) Extracts and Its Chlorogenic Acid Isomer Component Improve Glucolipotoxicity-Induced Hepatic Lipid Accumulation via Downregulating miR-34a and Decreased Inflammation

doi: 10.3390/nu14224808

Figure Lengend Snippet: MLE and nCGA reduced miR-34a expression in OH−induced HepG2 cells. ( A ) HepG2 cells were co-treated with OH, MLE (3 mg/mL), and nCGA (50 μM) at 37 °C for 24 h. The level of miR-34a was analyzed using Real-Time PCR. ( B ) Transfection with miR-34a mimics or inhibitors in the HepG2 cells were measured by Western blotting to detect SIRT1, PPAR-α, and p-AMPK/AMPK. Values are the mean ± SD of three independent experiments. Data were represented using the Student’s t -test. # p < 0.05 vs. control group. * p < 0.05 vs. the OH group.

Article Snippet: Monoclonal antibodies against phosphor-AMPK (#2535), FASN (#3180) and α-tubulin were obtained from Cell Signaling Technology Inc (Beverly, MA, USA).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Transfection, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Neochlorogenic Acid Attenuates Hepatic Lipid Accumulation and Inflammation via Regulating miR-34a In Vitro

doi: 10.3390/ijms222313163

Figure Lengend Snippet: The effect of 5-CQA on miR-34a expression and the effect of miR-34a on lipid accumulation. ( A ) The cells were co-treated with OA and 5-CQA for 24 h. The level of miR-34a was analyzed as described in the text. Treatment with 5-CQA decreased the expression of miR-34a. The results are presented as mean ± SD from three independent experiments for each group. # p < 0.05, in relation to the control group. * p < 0.05, in relation to the OA-induced group. Ctrl, control; OA, oleic acid. ( B ) After transfection with miR-34a mimic or inhibitor, the total cell lysate (protein) was analyzed using immunoblotting against SIRT1, phosphor-AMPK, AMPK, and actin antibody. The data are presented as mean ± SD. # p < 0.05, in relation to the negative control group. ( C ) After miR-34a inhibitor transfection, cells were treated with or without OA for 24 h. The intracellular lipid was stained with Oil Red O and the representative photo was taken. The quantification was determined using colorimetric assay as described in the text. The data are presented as the mean ± SD of three samples of each group. # p < 0.05, in relation to the negative control group. * p < 0.05, in relation to the OA-treated group.

Article Snippet: Cell Signaling Technology Inc (Beverly, MA, USA) provided antibodies against phosphor-AMPK (#2535) and FASN (#3180).

Techniques: Expressing, Transfection, Western Blot, Negative Control, Staining, Colorimetric Assay