Comparative Effect of Aqueous and Methanolic Bupleuri Radix Extracts on Hepatic Uptake of High-Density Lipoprotein and Identification of the Potential Target in HFD-Fed Mice

Figure 3

Flow cytometry, microscopy, and real-time imaging of DiI-HDL uptake by HepG2 cells. (a) The real-time imaging station IncuCyte ZOOM recorded HDL uptake by HepG2 cells at 0 h, 1 h, 2 h, 3 h, 4 h, and 5 h after exposure to DiI-HDL. (b) Fluorescent images of DiI-HDL uptake (40x) were obtained by Olympus BX53 after 24 hours of exposure to 10 mg/mL aqueous extract or methanolic extract of Bupleuri Radix. (c) FACS histograms of the fluorescence signal related to DiI-HDL uptake in 24 h cultured fibroblasts after treated with 5 mg/mL, 10 mg/mL, or 15 mg/mL aqueous Bupleuri Radix extract, respectively, from the left to right. (d) FACS histograms of the fluorescence signal related to DiI-HDL uptake in 24 h cultured fibroblasts after treated with 5 mg/mL, 10 mg/mL, or 15 mg/mL methanolic Bupleuri Radix extract, respectively, from the left to right. (e) The aqueous extract induced drastic increase of HDL uptake in vitro, while the methanolic extract reduced it. ; compared to the control group (t-Student test, unpaired two-tailed). Cells were incubated with DiI-HDL for 5 h before the harvesting times and prepared for FACS analysis as reported in Section 2. Each experiment was carried out at least twice in triplicate. At least 10000 cells for each group were analysed.

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