In VitroScreening for Antihepatic Steatosis Active Components within Coptidis Rhizoma Alkaloids Extract Using Liver Cell Extraction with HPLC Analysis and a Free Fatty Acid-Induced Hepatic Steatosis HepG2 Cell Assay

Abstract

A high-throughput method was developed and applied to screen for the active antihepatic steatosis components within Coptidis Rhizoma Alkaloids Extract (CAE). This method was a combination of two previously described assays: HepG2 cell extraction with HPLC analysis and a free fatty acid-induced (FFA) hepatic steatosis HepG2 cell assay. Two alkaloids within CAE, berberine and coptisine, were identified by HepG2 cell extraction with HPLC analysis as high affinity components for HepG2. These alkaloids were also determined to be active and potent compounds capable of lowering triglyceride (TG) accumulation in the FFA-induced hepatic steatosis HepG2 cell assay. This remarkable inhibition of TG accumulation (P < 0.01) by berberine and coptisine occurred at concentrations of 0.2 μg/mL and 5.0 μg/mL, respectively. At these concentrations, the effect seen was similar to that of a CAE at 100.0 μg/mL. Another five alkaloids within CAE, palmatine, epiberberine, jateorhizine, columbamine, and magnoline, were found to have a lower affinity for cellular components from HepG2 cells and a lower inhibition of TG accumulation. The finding of two potent and active compounds within CAE indicates that the screening method we developed is a feasible, rapid, and useful tool for studying traditional Chinese medicines (TCMs) in treating hepatic steatosis.

Advances in Chemistry and Bioactivity of Magnoflorine and Magnoflorine-Containing Extracts

“…The tested extract at the concentration of 12.5 to 100.0 µg/mL had a triglycerides reducing effect in a dose-dependent manner. MGN and other alkaloids from the extract like epiberberine, palmatine, or columbamine reduced triglyceride level in a statistically significant way only at the highest concentration applied (5.0 µg/mL) [71].…”

“…In vitro screening of alkaloid-rich liver cell extract obtained after the administration of Coptidis rhizoma and a free fatty acid-induced test for hepatic steatosis HepG2 cell assay was performed by Fan and colleagues [71]. Hepatic steatosis characterized by overmuch fat accumulation in hepatocytes is a serious clinical problem.…”

Tri-Herbal Medicine Divya Sarva-Kalp-Kwath (Livogrit) Regulates Fatty Acid-Induced Steatosis in Human HepG2 Cells through Inhibition of Intracellular Triglycerides and Extracellular Glycerol Levels

“…HepG2 cells were obtained from the National Centre for Cell Science, Pune, India (local repository for the American Type Cell Culture (ATCC)), proliferated within two passage numbers, and stored in liquid nitrogen at passage number 17. Cells were cultured in high glucose (4.5 gm/L) DMEM cell culture media (Gibco, Evansville, IN, USA) supplemented with 10% FBS (HiMedia, Mumbai, India) and 100 U/mL penicillin/streptomycin (Thermo Fisher Scientific, Waltham, MA, USA) and maintained at 37 °C and 5% CO 2 [ 63 ]. Cells were passaged at 70% confluence and used for assays based on the number of cells required in each experiment.…”

Coptisine fromCoptis chinensisexerts diverse beneficial properties: A concise review

“…During the past decades, many pharmacokinetic studies have been focused on the intracorporal process of formulas like SHXXT and BanXia Xiexin decoctions, the main constituents of which were all verified to include coptisine (Chemical structure showed in Figure ). In rat model administrated with SHXXT, coptisine was not metabolized in blood and could reach hepatic and liver cells as prototype form . To evaluate further coptisine metabolism in liver, a study conducted in zebrafish model demonstrated that coptisine has 4 metabolites and the main metabolic ways include demethylation, hydroxylation, sulphation and glucuronidation .…”

“…In rat model administrated with SHXXT, coptisine was not metabolized in blood and could reach hepatic and liver cells as prototype form. 17 To evaluate further coptisine metabolism in liver, a study conducted in zebrafish model demonstrated that coptisine has 4 metabolites and the main metabolic ways include demethylation, hydroxylation, sulphation and glucuronidation. 18 Moreover, coptisine was revealed to be the substrate of P-gp, 19 which to some extent, contributes to the low distribution of coptisine in multiple tissues.…”