ABSTRACT
To explore the regulatory effect and relevant mechanisms of the fraction of Hedyotis diffusa and Scutellaria barbata herb couple(YDW11) on polarization of macrophage between M1/M2 phenotypes.RAW264.7 cells were induced with LPS/IFN- or IL-4/IL-13 to establish M1 or M2 macrophage cell model. MTT assay was used to measure the cell cytotoxicity of YDW11. Griess reaction was used to detect the changes of nitrite accumulation in the cell supernatant. Trans-well assay was used to measure the migration capability. QRT-PCR was used to assay mRNA expressions of iNOS, IL-1, Arg-1 and MR. Western blot was used to detect the effect of YDW11 on iNOS and Arg-1 protein expressions. Taqman MicroRNA RT-PCR was used to detect the effect of YDW11 on miR155 expression under M1 and M2 phenotype conditions. In addition, MS-UPLC assay was carried out to identify the constituents in YDW11. The results showed that the ethyl acetate of H. diffusa and S. barbata extracted in 1:1 ratio with water (YDW11) showed the activity in suppressing the nitrite content in M1 macrophages without cytotoxicity. YDW11 also inhibited the migration of breast cancer cells with the help of M2 macrophages by blocking their polarization towards M2. YDW11 decreased iNOS, IL-1, Arg-1and MR mRNA expressions and iNOS and Arg-1 protein expressions. YDW11 down-regulated miR155 expression in M1 phenotype, and up-regulated miR155 expression in M2 phenotype. Based on MS-UPLC,four compounds were identified in YDW11, including 4'-hydroxyacetophenone, scutellarin, luteolin and apigenin. YDW11 inhibited M1/M2 phenotypes of macrophages by regulating the expression of miR155.
ABSTRACT
Aims Tocomparethepharmacokineticsof ephedra alkaloids,cinnamic acid and cinnamic alcohol in Ephedrae, Cinnamomi and Ephedrae-Cinnamomi herb couple through UPLC-MS/MS in rats respective-ly,and to investigate the effect of combination on phys-iologicaldisposition.Methods Plasmasampleswere collected at different times after oral administration of Ephedrae,Cinnamomi and Ephedrae-Cinnamomi herb couple extracts.The concentrations of ephedra alka-loids,cinnamic acid and cinnamic alcohol in plasma samples were determined by UPLC-MS/MS.DAS 3. 0 was used to calculate pharmacokinetic parameters.The differences of samples in two groups were conducted with univariate statistical analysis using SPSS 13. 0. Results ComparedwithEphedraegroup,theCmaxof norephedrine hydrochloride, norpseudoephedrine hydrochloride,ephedrine hydrochloride,pseudoephed-rine hydrochloride,and methylephedrine hydrochloride in Ephedrae-Cinnamomi herb couple group were signif-icantly greater (P<0. 05 );the AUC0-t of norpseudo-ephedrine hydrochloride was significantly greater (P<0. 05 );the MRT0-t of norephedrine hydrochloride, phedrine hydrochloride, pseudoephedrine hydrochlo-ride,and methylephedrine hydrochloride were signifi-cantly less (P <0. 05 );the T1/2z of norephedrine hydrochloride,phedrine hydrochloride,and methyl-ephedrine hydrochloride were significantly less (P <0. 05 ).The AUC0-t and MRT0-t of cinnamic acid and cinnamic alcohol were significantly greater than those inCinnamomigroup(P<0.05).Conclusion The combination of Ephedrae and Cinnamomi improves the absorption concentration of five ephedra alkaloids, Slows down the elimination of norephedrine hydrochlo-ride,phedrine hydrochloride,pseudoephedrine hydro-chloride,and methylephedrine hydrochloride,and in-creases the bioavailability of cinnamic acid and cin-namic alcohol.
ABSTRACT
This study was aimed to observe the antipyresis and cholagogue effect of theFructus Gardeniae-Fructus Forsythiaeherb couple in order to explore the possible pharmacodynamic mechanism. Dry yeast suspension was subcutaneously injected into the back of rat to establish the fever model. The freeze-dried powders ofF. Gardeniae,F. Forsythiae, andF. Gardeniae-F. Forsythiaeherb couple were prepared. They were dissolved in the water for intragastric administration. SD rats were randomly divided into the normal group, model group, positive control group and Chinese medicine group. The Chinese medicine group was subdivided into the group ofF. Gardeniae,F. Forsythiae, andF. Gardeniae-F. Forsythiaeherb couple. The concentration of herbal water extract was 10 mL·kg-1 (which equaled to 3 g·kg-1 of a single crude herb). The concentration of positive control was 10 mL·kg-1. Intragastric administration of equal amount of normal saline was given to the blank group and the model group. Except rats in the normal group, rats in other groups were subcutaneously injected with 10 mL·kg-1 of 15% dry yeast suspension on the back of to establish the fever model. Electronic thermometer was used to record the body temperature of rats at 0, 1, 4, 5, 6, 7 and 8 h after the injection, respectively. Meanwhile, bile was collected from 1-1, 1-2, 2-4, 4-6, 6-8, 8-12, 12-24 h, respectively. Observation was given on changes of body temperature and bile amount of rat. The results showed thatF. Gardeniae,F. Forsythiae, andF. Gardeniae- F. Forsythiaeherb couple had certain effect to reduce the body temperature of rats. The temperature-reducing effect of the combination of both herbs was better than a single herb. TheF. Gardeniae -F. Forsythiae herb couple can reduce the body temperature of fever rat (P < 0.05, orP < 0.01). It had a little effect on the body temperature of normal rat. TheF. Gardeniae - F. Forsythiaeherb couple can promote the bile secretion, which was better than the single using of F. Gardeniae (P < 0.05). It was concluded that theF. Gardeniae - F. Forsythiaeherb couple had better temperature-reducing effect than the using of a single herb; however, there was no significant difference. But it had obvious effect on the promotion of bile secretion, which indicated the strengthening of cholagogue effect.