RESUMO
An ultra-high performance liquid chromatography method for the determination of 8 constituents in Qingzao Jiufei Decoction was established and the basis of related chemical substances with antioxidant activity in Qingzao Jiufei Decoction was explored. The separation was performed on a Waters Cortecs RP Shield C18 (150 mm × 2.1 mm, 1.6 μm) using UHPLC-DAD as the mobile phase was water (containing 0.1% phosphoric acid) – acetonitrile with flow rate of 0.30 mL·min-1 by gradient elution ① determining 5 constituents (amygdalin, liquiritin, liquiritin apioside, rutin and isoquercitrin) at the wavelength of 210 nm, 237 nm and 358 nm. Under gradient elution ②, 3 constituents (glycyrrhizin, glycyrrhizic acid and sesamin) were determined at the wavelength of 210 nm and 265 nm. The IC50 of 10 batches of Qingzao Jiufei Decoction scavenging 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS+) free radicals obtained through test and Probit model was analyzed for correlation with the contents of 8 constituents. The established methods had a good linear relationship (r > 0.999), good repeatability and stability. The recovery rate was between 82.8% and 112.4%. In a series of concentration range, the higher the concentration of Qingzao Jiufei Decoction, the stronger the free radical scavenging effect. There was a significant correlation between the content of rutin and glycyrrhizic acid and the IC50 of scavenging free radicals. The content determination methods established in this experiment provide a basis for a reasonable and scientific evaluation of the quality of Qingzao Jiufei Decoction. Qingzao Jiufei Decoction has antioxidant activity, which is significantly positively correlated with the content of rutin and glycyrrhizic acid.
RESUMO
Objective To explore the effect of Qingzao Jiufei Decoction (QJD) and its decomposing agent on the expression of Bax, Bcl-2, and Caspase-3 apoptosis protein in MP infection, in order to determine the effect target of QJD and its decomposing agent. Methods A total of 120 balb/c mice were randomly divided into normal group (group A), model group (group B), QJD group (group C), QJD group I decomposition agent (Group D), QJD group II decomposition agent (Group E), and azithromycin group (Group F), 20 rats in each group. Except the normal group, the other five groups were infected with MP by using the nose drop method. The ultrastructure and apoptosis of lung tissue were observed by transmission electron microscope. The expression of Bcl-2, Bax and Caspase-3 protein in lung tissue was detected by Immunohistochemical SP and Western blot method. The expression of Caspase-3 m-RNA was detected by qPCR method. Results After MP infection, inflammation changes can be observed in the lung tissue of mice, thickening of the alveolar wall, destruction of alveolar epithelial cells, cell infiltration, and changes in the characteristics of apoptosis. The expression of Bcl-2, Bax, Caspase-3 protein in the lung tissue of mice infected with MP was significantly increased, but the ratio of Bcl-2/Bax decreased significantly. Compared with the group B, the expression of Bcl-2 in group C, D, and F increased, the ratio of Bcl-2/Bax increased significantly, and the expression of Bax and Caspase-3 decreased. The difference of expression between group E and group B was not obvious. The results of Caspase3 mRNA detection showed that the expression of group C, group D and group F was lower than that of model group, and the change of group E was not obvious. Conclusion QJD can inhibit the cell apoptosis induced by MP infection, and Bax, Bcl-2 were one of its effect target, in which I decomposition agent plays a major role.