Bioactivity of five components of Chinese herbal formula Jiangzhi granules against hepatocellular steatosis

OBJECTIVE： This study aims to evaluate the bioactivity of five components of the traditional Chinese medicine complex prescription Jiangzhi granules against hepatocellular steatosis. METHODS： The five major components, including protopanaxadiol, tanshinone IIA, emodin, chlorogenic acid, and nuciferine, were extracted from Jiangzhi granules. Their cytotoxicity was assessed to determine the safe dose of each component for HepG2 cells. HepG2 cellular steatosis was induced using 1 mmol/L of free fatty acids （FFAs） for 24 h, and then treated with each component at high, intermediate, and low doses （500, 50, and 5 μmol/L）, respectively for another 24 h. The effects on HepG2 steatosis were observed directly under optical phase microscopy, or through oil red O staining and Nile red assays. In addition, the levels of reactive oxygen species （ROS） in the steatotic HepG2 cells with and without high-dose protopanaxadiol treatment were measured using fluorescent dye 2＇,7＇-dichlorodihydrofluorescein diacetate staining. RESULTS： No obvious cytotoxicity was observed in the HepG2 cells incubated with each of the five components at up to 500μmol/L. At 24 h after incubation with FFAs, the HepG2 cells swelled and many lipid droplets accumulated. The lipid content was attenuated after 24 h of incubation with protopanaxadiol, tanshinone IIA, and emodin at 500 or 50 μmol/L （P 〈 0.05）, especially with 500 μmol/L protopanaxadiol （P 〈 0.01）. In addition, the ROS level was elevated in steatotic cells, but decreased after intervention with 500μmol/L protopanaxadiol （P 〈 0.05）. CONCLUSION： Protopanaxadiol, tanshinone IIA, and emodin alleviate hepatocellular steatosis in a dose-dependent manner, and oxidative stress regulation may partially contribute to the effects of protopanaxadiol. ：

Quantitative determination of multi-class bioactive constituents for quality control of Yiqi Jiangzhi Granules

Objective:To establish a reliable and sensitive method for evaluating quality of Yiqi Jiangzhi Granules(YQJZG).Methods:Ultra performance liquid chromatography electrospray ionization tandem mass spectrometry(UPLC-ESI-MS/MS)was employed for simultaneous determination of eight marker components.Separation was performed on an AQUITY UPLC;HSS T3 column,the mobile phase consisted of acetonitrile as the organic phase and 0.1%(volume percentage)formic acid as the aqueous.Eight marker components,ginsenoside Rg1(GRg1),ginsenoside Re(GRe),ginsenoside Rb1(Gb1),typhaneoside(TEO),isorhamnetin-3-O-neohespeidoside(IN),hesperidin(HPD),aurantio-obtusin-6-O-β-D-glucoside(AG)and curcumin(CCM),were detected by multiple reaction monitoring(MRM)mode.The Chinese Pharmacopoeia(2020 edition)was regarded as the guidance document for this method validation.Results:The method showed good linearity(R^(2)≥0.9990).The relative standard deviation(RSD)values for the instrument precision,intermediate precision and repeatability were less than 2.91%,2.88%,and 3.54%,respectively.The average recovery varied from 91.08%to 103.89%,with RSD below 3.81%.Sample solutions were found to be stable within 24 h at 4℃(RSD<2.85%).Eight marker components were successfully determined from three batches of YQJZG.Conclusion:The proposed UPLC-ESI-MS/MS method was found to be simple,fast and sensitive,and can be used for the routine quality assessment of YQJZG.Simultaneously,this method may provide a new and powerful tool of quality control for other traditional Chinese medicine analogous formulae.