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Objective: To establish a study method for discovering quality marker of herb Meconopsis integrifolia (Non flower parts) based on spectrum-effect relationship for its quality control. Methods: UPLC fingerprints and UPLC-ESI-Q-TOF-MS/MS method were used to identify the chemical components of 10 batches of HMI extracts. The total antioxidant capacity, ABTS free radical scavenging ability, DPPH free radical scavenging ability and superoxide anion free radical suppression ability of 10 batches of sample were determined. Principal component analysis was used to screen antioxidant capacity index for HMI. Grey relational analysis was used to analyze the correlation between antioxidant capacity index and common peak of fingerprint spectrum. Then, quality marker could be preliminarily discovered by comprehensive analysis associated with antioxidant capacity index and efficacy-related constituents. Finally, the constituents were separated by HSCCC. Results: A total of 29 common peaks were identified from fingerprints of HMI extracts. 18 constituents were identified by UPLC-ESI-Q-TOF-MS/MS analysis, including 16 flavonoids and two alkaloids. DPPH scavenging ability was screened by principal component analysis which could reflect the antioxidant capacity of HMI extracts. Grey relational analysis showed that the correlations between the 29 common peaks and DPPH scavenging ability were all greater than 0.5. The quality markers of HMI were quercetin 3-O-β-D-glucopyrannosy-(1→6)-β-D-glucopyranoside and quercetin 3-O-[2'''-O-acetyl-β-D-galactopyranosyl-(1→6)-β-D-glucopyranoside] by comprehensive analysis. Among them, quercetin 3-O-[2'''-O-acetyl-β-D-galactopyranosyl-(1→6)-β-D- glucopyranoside] is a new compound. Conclusion: Study on the quality marker of HMI based on the spectrum-effect relationship is of great significance for elucidating the pharmacodynamic material basis, screening core quality marker related to medicinal effect, and ensuring the safety and rational application of traditional medicines.
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Objective Coptidis Rhizoma (CR), a widely used traditional Chinese herbal medicine, is commonly believed to be non-toxic. However, little is known about its cytotoxicity and relevant mechanisms at cellular and genetic levels. The present study was conducted to explore the cytotoxicity of CR and its mechanisms related to cell cycle arrest, DNA damage, cell apoptosis, and mitochondrial membrane potential in L929 murine fibroblast cells. Methods The cells were cultured and treated with different concentration of CR aqueous extract for 24 h. Cell viability was determined by CCK-8 method, morphological changes, and mitochondrial membrane potential were observed with an inverted microscope, cell cycle and cell apoptosis were examined by flow cytometry and DNA damages were detected by comet assay. Results Our results showed that cell viability was significantly decreased in a dose-dependent manner when concentration was higher than 0.2 mg/mL. A concentration above 1 mg/mL altered the cells morphology. Each DNA damage indicator score increased in the groups with the concentration of above 0.1 mg/mL. Cells at G2/M phase, cell apoptosis and mitochondrial membrane potential changed in the 2 mg/mL group. Conclusion Overall, our study suggests that CR at a high dosage exhibits cytotoxicity on L929 cells, which is likely to be the consequences of cell cycle arrest, DNA damage, cell apoptosis and mitochondrial membrane potential reduction.
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The influence on 10 kinds of ginsensides of different processed methods of Panacis Quinquefolii Radix was discussed. White Panacis Quinquefolii Radix (sliced and dried at -80 °C), red Panacis Quinquefolii Radix( steamed, sliced and dried at -80 °C) and commercial Radix Panacis Quinquefolii (dried by electric blast air) processed by different methods. HPLC-PDA-ESI- MS method was established before by our team. Ten kinds of ginsenosides of them were determined. The content of total ginsenosides were as follow: commercial Panacis Quinquefolii Radix > white Panacis Quinquefolii Radix > red Panacis Quinquefolii Radix. Compared with white Panacis Quinquefolii Radix, the content of Re, Rc, Rb3 and Rb2 of Red Radix Panacis Quinquefolii decreased but increased that of Rg,, Rb1. Both Rg2 and Rg, were not found in white Panacis Quinquefolii Radix and commercial Panacis Quinquefolii Radix by PDA detector, and low response in ESI-MS, while red Panacis Quinquefolii Radix was to the high content that of 0. 027% and 0.040 1%. The constituent of RA0 of red Panacis Quinquefolii Radix was higher than the other two. After Panacis Quinquefolii Radix processed, the kind and content of ginsensides were significantly changed. The constituent of some kinds of ginsensides was increased and some decreased. Rf was not found in all Panacis Quinquefolii Radix samples which were consistent with the former documents.
Subject(s)
Chemistry, Pharmaceutical , Methods , Ginsenosides , Chemistry , Mass Spectrometry , Panax , Chemistry , Plant Extracts , Chemistry , Plant Roots , ChemistryABSTRACT
OBJECTIVE@#To investigate the effects of inflammation cytokines, (FK506) and cyclosporine (CSA) on albumin secretion, and the effects of FK506 and CSA on the IL-6 induced suppression of albumin synthesis in cultured human hepatocytes.@*METHODS@#Human hepatoma cell lines (HepG2 cells) were separately cultured with IL-6, IL-2 and IL-10 (0 approximately 10 microg/L) and FK506, CSA (0 approximately 100 microg/L) for 48 h. In another experiment, HepG2 cells were stimulated with different doses of FK506 and CSA (0 approximately 10 microg/L) in the presence of IL-6 (5 microg/L) for 48 h. Albumin levels in the supernatant of all groups were measured by radioimmunoassay (RIA). The concentration of LDH secreted by cells stimulated with FK506 and CSA were detected with spectrophotometry.@*RESULTS@#For cultured HepG2 cells, IL-6 significantly decreased albumin levels in a dose-dependent manner (P 0.05). FK506 obviously decreased LDH levels in the supernatant (P 0.05).@*CONCLUSION@#IL-6 but not IL-2 and IL-10 suppressed the production of hepatic albumin in vitro. FK506 protected against the suppression of hepatic albumin synthesis caused by IL-6, suggesting its potential role in improving hypoalbuminaemia in immune glomerulonephritis.