Screening nephroprotective compounds from cortex Moutan by mesangial cell extraction and UPLC

Abstract

A method for screening nephroprotective compounds in cortex Moutan, a common traditional Chinese medicine (TCM) in treating diabetic nephropathy with renal mesangial cell extraction and ultra performance liquid chromatography technique was described in this paper. We hypothesize that the compounds which bind to cell membranes under pathological conditions may be the bioactive compounds in TCMs. Mesangial cells were cultured in medium containing 5 mM (physiological, NG) or 30 mM (pathological, HG) glucose for 48 h and then incubated with cortex Moutan extract. After the unbound substances were washed off, the cell membrane-bound compounds were dissociated and concentrated by an SPE column. By comparing the chromatograms of NG and HG cultured-cell extractions and cortex Moutan extract, three compounds bound to both NG and HG-cultured mesangial cells were identified as paeoniflorin, pentagalloylglucose (PGG) and paeonol. In vitro studies showed that paeoniflorin, PGG and paeonol reduced the activity of nicotinamide-adenine dinucleotide phosphate oxidase (NADPH) activity, and decreased the level of reactive oxygen species (ROS), transforming growth factor-β1 (TGF-β1) and fibronectin in high glucose cultured mesangial cells. The results indicate that paeonol, paeoniflorin and PGG may be the nephroprotective compounds from cortex Moutan. This study is expected to provide a more reliable and effective method for screening bioactive compounds from the complex TCM systems.

DJC Suppresses Advanced Glycation End Products‐Induced JAK‐STAT Signaling and ROS in Mesangial Cells

“…NADPH oxidase activity was measured by lucigenin-enhanced chemiluminescence as previously described [20]. GMCs (5 × 10 4 cells in 2 mL) were seeded on 12-well plates in quadruplicate and cultured with AGEs (250  μ g/ml) for 48 h in the presence or absence of graded concentrations of DJC or AG490.…”

Paeonol Ameliorates Diabetic Renal Fibrosis Through Promoting the Activation of the Nrf2/ARE Pathway via Up-Regulating Sirt1

“…( Sun et al, 2008 ; Chen et al, 2012 ; Zhang et al, 2013 ; Gong et al, 2017 ). Recent studies reveal that PA treatment reduces blood glucose level in STZ-induced diabetic rats ( Liu et al, 2013 ) and decreases content of FN and TGF-β1 in the supernatant of HG-cultured mesangial cells ( Sun et al, 2015 ) separately, which indicate the potential renoprotective effect of PA. However, the molecular mechanism by which PA exerts renoprotective activity remains unclear.…”

Screening active components from Rubus amabilis for pancreatic β-cells protection

“…Intracellular ROS generation was determined using the fluorescent probe DCFH-DA as described previously (Sun et al 2015). MIN6 cells were seeded in a 96-well plate and treated with 0.5 mM PA in the presence or absence of PCs at different concentrations for 24 h. Then, the cells were exposed to DCFH-DA (10 lM) in the dark for 30 min at 37 C. The fluorescence was read at Ex 485/Em 530 nm on a Synergy HI Multi-Mode Microplate Reader.…”

“…Cell membrane chromatography (CMC), which detects the bound components from cell extractions via liquid chromatography, is a convenient, specific, and time-saving technique for screening active components from complicated herbal medicines. Some potential active components in Chinese medicine have been screened using various cell extractions, including mesangial cell (Sun et al 2015), hepatocyte (Hong et al 2012), epithelial cell (Shen et al 2014), and macrophage (Yu et al 2007) extraction. In this paper, the potential active components in R. amabilis were screened using pancreatic b-cell (MIN6 cell line) extraction, and the protective effects of the screened components were further examined using palmitate-treated MIN6 cells.…”