Comparison of Effect of Chijian and Gastrodiae Rhizoma on Regulating Immune Function and Intestinal Microflora / 中国实验方剂学杂志

ABSTRACT

Objective:To compare the contents of adenosine， gastrodin， <italic>p</italic>-hydroxybenzyl alcohol， <italic>p</italic>-hydroxybenzaldehyde， parisinin B and parisinin A in Chijian （the aerial part of <italic>Gastrodia elata</italic>） and Gastrodiae Rhizoma， and compare their effects on immune function and intestinal microflora， evaluating whether it is necessary to study and develop Chijian. Method:The contents of these six constituents were determined by ultra performance liquid chromatography （UPLC）， the mobile phase was 0.1% formic acid aqueous solution （A）-acetonitrile （B） for gradient elution （0-4 min， 0.5%B； 4-5 min， 0.5%-2%B； 5-10 min， 2%-15%B； 10-12 min， 15%-20%B； 12-15 min， 20%-95%B； 15-17 min， 95%B； 17-17.5 min， 95%-0.5%B； 17.5-20 min， 0.5%B）， the flow rate was 0.5 mL·min^-1， the detection wavelength was 270 nm. The difference of pharmacological activity of water extracts of Chijian and Gastrodiae Rhizoma was compared， the clearance index， corrected clearance index and peripheral blood were measured in mice model with low immune function induced by cyclophosphamide， B lymphocyte proliferation was determined by lymphocyte transformation test <italic>in vitro</italic>， intestinal microflora was analyzed by 16S rDNA technology and bioinformatics was conducted. Result:The total contents of these six components in powder and ethanol extract of Chijian were higher than that of Gastrodiae Rhizoma， but the total contents of these six components in their water extract were similar， and the total contents of gastrodin and <italic>p</italic>-hydroxybenzyl alcohol met the requirements of the 2020 edition of <italic>Chinese Pharmacopoeia</italic>. Compared with the blank group， the clearance index of immunocompromised mice was significantly increased in the middle-dose （10 g·kg^-1） group of Chijian water extract， middle- and low-dose （10， 5 g·kg^-1） groups of Gastrodiae Rhizoma water extract （<italic>P</italic><0.05）， the levels of erythrocyte and hematocrit in peripheral blood were significantly increased in the high-dose （20 g·kg^-1） groups of water extracts of Chijian and Gastrodiae Rhizoma （<italic>P</italic><0.05， <italic>P</italic><0.01）， water extract of Gastrodiae Rhizoma with concentration of 400 g·L^-1 and the water extract of Chijian with the concentration of 100 g·L^-1 could promote the proliferation of B lymphocytes induced by lipopolysaccharide. Studies on intestinal microflora showed that compared with the blank group， at the phylum level， the water extracts of Chijian and Gastrodiae Rhizoma increased the relative abundance of Bacteroidetes and decreased the relative abundance of Firmicutes， at the genus level， they increased the relative abundance of <italic>Prevotellaceae</italic>_UCG-001 and <italic>Ruminococcaceae</italic>_UCG-005， and decreased the relative abundance of <italic>Anaerotruncus</italic>， unclassified_<italic>f</italic>_<italic>Erysipelotrichaceae</italic> and<italic> Candidatus</italic>_<italic>Stoquefichus</italic>.<italic> </italic>These intestinal bacteria were related to the immune system， cell proliferation， and metabolism regulation. Conclusion:The total contents of 6 components in the powder， the ethanol and the water extracts of Chijian are higher than or close to those of the corresponding samples of Gastrodiae Rhizoma， the pharmacological activity of Chijian water extract is similar to that of Gastrodiae Rhizoma water extract， indicating that Chijian is worthy of further research and development.