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Using the concepts and methods of epigenetics and metabolomics, to investigate the overall action molecular mechanism of Chrysanthemi indici C (CIC), the anti-hepatitis B virus (HBV) active extracts from Flos chrysanthemi indici. The inhibitory effects of CIC on proliferation and hepatitis B surface antigen (HBsAg), hepatitis B envelope antigen (HBeAg) and HBV-DNA of HepG2.2.15 cells were detected by CCK-8 and antigen kit. The DNA methyltransferases (DNMTs)/ten-eleven-translocation-2 (TET2) equilibrium was detected by ELISA. Illumina 850K methylation chip, pyrosequencing and qPCR were used to determine the action pathway and target of CIC by GO and KEGG analysis. Cell metabolites were extracted with 80% methanol, and the changes of differential metabolites, differential metabolic pathways and cell microenvironment were detected by LC-MS and other metabolomics methods. The results showed that CIC could inhibit the proliferation, HBsAg, HBeAg and HBV-DNA of HepG2.2.15 cells obviously, down-regulate DNA methyltransferase 1 (DNMT1), DNA methyltransferase 3a (DNMT3a) and DNA methyltransferase 3b (DNMT3b), up-regulate TET2, and restore the balance of DNMTs/TET2. The action targets of CIC were phospholipase C gamma 2 (PLCG2), phosphoinositide-3-kinase regulatory subunit 3 (PIK3R3), 1-acylglycerol-3-phosphate O-acyltransferase 2 (AGPAT2), 5-hydroxytryptamine receptor 2B (HTR2B), nerve growth factor (NGF), mainly involved in lipid metabolism, inflammation mediated regulation of transient receptor potential (TRP), phospholipase D signaling and advanced glycation end product-receptor for AGE (AGE-RAGE) signaling in diabetic complications pathways. CIC could significantly affect fatty acid metabolism and had great influence on phenolic acid, alkaloid and lipid metabolites in cell microenvironment. These results suggest that the action mechanism of CIC may be the synergistic action of multiple pathways and multiple targets, including related inflammatory pathways, immune pathways and lipid metabolism, through regulating epigenetic expression balance and restoring the balance of cell microenvironment.
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Objective To evaluate the antimicrobial activity and establish HPLC fingerprint of flavonoids in Flos Chrysanthemi Indici. Methods Microdilution method was carried out to determine the minimum inhibitory concentration (MIC) value of total flavonoids and linarin against eleven kinds of bacteria and three fungi. HPLC fingerprint was obtained on Diamonsil C18 (4. 6 mm × 250 mm, 3 μ,m) with gradient elution consisting of acetonitrile (A) and 0. 05% phosphoric acid (B), detective wavelength was at 326 nm and flow rate 1. 0 ml/min n he room temperature. The data were analyzed by Computer Aided Similarity Evaluation software. Results MIC values of total flavonoids and inarin against Candida lusitaniae were 61. 6 μg/ml and 18. 0 μg/ml, respectively. Furthermore, fifteen characteristic peaks were indicated n fingerprint chromatography, and eight peaks were identified as flavonoids. Conclusion Total flavonoids and he inarin n Flos Chrysanthemi Indici exhibited better antimicrobial activity against C. lusitaiae. The established HPLC fingerprint method s accurate and reproducible, and can be used for the basic research and quality control of active flavonoids in Flos Chrysanthemi Indici.
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AIM:To optimize the technical parameters indica-daisy dropping pill(Flos chrysanthemi indici) through controlling the influencing factors. METHODS:To take weight variation of pills,comprehensive quality and disintegration time limited as index,the above factors were observed by orthogonal test. RESULTS:Good technological parameters of indica-daisy dropping pill were as follows PEG6000∶PEG400=54∶6 as matrix,dimethicone as refrigerant.Temperature of drug fluids of 70 ℃,internal and external diameter of dropper within 6.2 mm and 9.0 mm,proportion of drug and matrix(1∶3),30 dropping per minute.The weight variation of pills was small,type quality was good and disintegration time limited was short.The water absorbability was smaller than that of granules. CONCLUSION:The finished products are of good quality and appropriate for mass production.