Animals ; Cattle ; Fatty Liver ; Inflammation ; Liver ; Liver Extracts ; Non-alcoholic Fatty Liver Disease ; Rats

Objective:The differences of chemical compositions and pharmacological activities between the core and pulp of Phyllanthi Fructus were investigated by chemical analysis and in vitro test to explore the effect of the core on the quality of this medicinal material. Method:Literature， medicinal material standards and market research on the appearance of Phyllanthi Fructus were conducted based on existing databases. Ultra-high performance liquid chromatography-quadrupole-electrostatic field orbital trap high resolution mass spectrometry （UPLC-Q-Orbitrap HRMS） was used to identify the constituents of the core and pulp. The analysis was performed on Thermo Scientific Accucore C₁₈ column （2.1 mm×100 mm， 2.6 μm） with the mobile phase of 0.1% formic acid aqueous solution （A）-methanol （B） for gradient elution （0-25 min， 5%B； 25-30 min， 5%-95%B； 30-35 min， 95%-5%B）， the flow rate was 0.2 mL·min^-1， heating electrospray ionization （HESI） was adopted with positive and negative ion modes， and the scanning range was m/z 100-1 500. High performance liquid chromatography （HPLC） was used to determine the contents of gallic acid， corilagin， chebulagic acid and ellagic acid in the core and pulp of Phyllanthi Fructus. Analysis was performed on Welchrom C₁₈ column （4.6 mm×250 mm， 5 μm） with mobile phase of methanol （A）-0.05% phosphoric acid aqueous solution （B） for gradient elution （0-6 min， 5%A； 6-15 min， 5%-7%A； 15-20 min， 7%-15%A； 20-25 min， 15%-21%A； 25-31 min， 21%-22%A； 31-41 min， 22%A； 41-47 min， 22%-28%A； 47-51 min， 28%-32%A； 51-57 min， 32%-38%A； 57-70 min， 38%-45%A； 70-80 min， 45%-65%A； 80-85 min， 65%-5%A）， the detection wavelength was set at 270 nm. The antibacterial effects of the core and pulp of Phyllanthi Fructus on Escherichia coli and Staphylococcus aureus were investigated by filter paper method， and their antioxidant activities were compared by 1，1-diphenyl-2-picrylhydrazyl （DPPH） free radical scavenging assay. Result:A total of 47 compounds were identified in the core and pulp of Phyllanthi Fructus， mainly including tannins， flavonoids， phenolic acids， fatty acids， amino acids， organic acids， saccharides and glycosides， most of which were concentrated in the pulp， and the fatty acids in the core accounted for a higher proportion. The contents of gallic acid， corilagin， chebulagic acid， ellagic acid and other phenolic compounds in the pulp of 20 batches of Phyllanthi Fructus were much higher than those in the core. The results of antibacterial test showed that the core of Phyllanthi Fructus with different concentrations had no antimicrobial effect. The DPPH radical scavenging test showed that the antioxidant activity of the core ［half-inhibitory concentration （IC₅₀）=199.632 mg·L^-1］ was much less than that of the pulp （IC₅₀=12.688 mg·L^-1）. Conclusion:From the perspectives of polyphenol content， antibacterial and antioxidant activities， it is scientific to use Phyllanthi Fructus pulp in ancient and modern times， which may be to remove the secondary parts of Phyllanthi Fructus， so as to enhance the actual utilization rate and therapeutic effect of medicinal materials. In view of the large proportion of the core of Phyllanthi Fructus and its high content of fatty acids and other components， whether or not to use it remains to be further studied in clinical application.

Indigo Naturalis( IN) is mainly composed of 10% organic matter and 90% inorganic matter,with a poor wettability and strong hydrophobicity. Indigo,indirubin and effective ingredients are almost insoluble in water. And how it exerts its effect after oral administration still needs to be revealed. For this reason,this study put forward the hypothesis that " Indigo Naturalis forms a slightly soluble calcium carbonate carrier in a strong acid environment of gastric fluid,and organic substances are solubilized in the bile environment of intestinal fluid",and then verified the hypothesis. First,the dissolution apparatus was used to simulate the change process of IN in different digestive fluid,and the effects of low-dose and normal bile on the dissolution of inorganic substances and the release of organic substances were compared. After the surface morphology and element changes of IN in different digestive fluid were observed,it was found that bile is the key to promoting the dissolution of organic and inorganic substances in IN. Furthermore,the rat fever model induced by 2,4-dinitrophenol was used to study the antipyretic effect of IN in normal rats and bile duct ligation rats. It was found that the antipyretic effect of IN on normal rats was better than that of bile duct ligation rats. The above results indicated that after oral administration of IN,the calcium carbonate carrier was transformed into a slightly soluble state in acidic gastric fluid,and a small amount of organic matter was released. When IN entered the intestinal fluid mixed with bile,the carrier dissolved in a large amount,and indigo and indirubin were dissolved in a large amount,so as to absorb the blood and exert the effect. This study has a certain significance for guiding clinical application of IN. For patients with insufficient bile secretion( such as bile duct resection),oral administration with IN may not be effective and shall be paid attention.
Animals ; Bile ; Humans ; Hydrophobic and Hydrophilic Interactions ; Indigo Carmine ; Indigofera ; Plant Extracts ; Rats