RESUMO
OBJECTIVE: To study the immunoregulatory effect of an optimal Chinese herbal monomer compound, which consists of three monomers, namely, icariin, baicalin and Astragalus saponin I, in a mouse model of allergic rhinitis. METHODS: A mouse model of allergic rhinitis was established by intraperitoneal injection of ovalbumin and aluminum hydroxide gel suspension. The splenic lymphocytes of the mice were separated, cultured in 96-well plates and divided into three groups: control group, concanavalin A group and compound group. Splenic lymphocyte proliferation was detected by cell counting kit-8 method at different time points. Cell cycle distribution was observed by flow cytometry (FCM) also at different time points. The changes of intracellular calcium concentration of splenic lymphocytes were measured by fluorescence microplate reader after the cells were incubated with fluorescence probe Fluo-3/AM. RESULTS: The Chinese herbal monomer compound could inhibit cell proliferation induced by concanavalin A (P<0.01). And the inhibition presented a time-effect relationship. With extending of the action time, the inhibition rate gradually increased and reached peak at the 48th hour. FCM test revealed the fact that concanavalin A could promote cells to enter into the mitosis by reducing the percentage of cells in G0/G1 phases while increasing the percentage of cells in S and G(2)/M phases. Compared with the concanavalin A, the compound could increase the percentage of cells in G(0)/G(1) phases and at the same time reduce the percentage of cells in S and G(2)/M phases at different time points, with the effect most significant at the 24th hour (P<0.05 or P<0.01). The results of the test taken by the fluorescence microplate reader revealed that the fluorescence value of the concanavalin A group increased with time in the previous 24 h while the compound could reduce this trend obviously, thus reduce the intracellular calcium concentration (P<0.01). CONCLUSION: The Chinese herbal monomer compound can inhibit the proliferation of cultured splenic lymphocytes of mice with allergic rhinitis. The effects of the compound of lowering intracellular calcium concentration and arresting cell cycle at G(0)/G(1) phases from entering into S and G(2)/M phases are responsible for its antiproliferation activity.
Assuntos
Proliferação de Células/efeitos dos fármacos , Medicamentos de Ervas Chinesas/farmacologia , Imunomodulação/efeitos dos fármacos , Linfócitos/citologia , Rinite Alérgica Perene/imunologia , Animais , Células Cultivadas , Composição de Medicamentos , Medicamentos de Ervas Chinesas/química , Flavonoides/farmacologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina , Rinite Alérgica , Rinite Alérgica Perene/induzido quimicamente , Saponinas/farmacologia , Baço/citologia , Triterpenos/farmacologiaRESUMO
OBJECTIVE: To study the optimal combined ratio of baicalin, icariin and Astragalus saponin I from a Chinese herbal compound Biminne. METHODS: Firstly, a mouse model of allergic rhinitis was established by intraperitoneal injection of ovalbumin (OVA) and aluminum hydroxide gel suspension, and the effective dose range of baicalin, icariin and Astragalus saponin I was detected by 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide inner salt method. Secondly, 10 groups of combinations of baicalin, icariin and Astragalus saponin I assembled by U(10)(10(8)) form were employed to determine the optimal combination by means of analyzing of the inhibitory effect on the splenocyte proliferation. Finally, the effects of each effective ingredient and the optimal combination were compared by observing the splenocyte proliferation, the contents of interleukin-4 (IL-4), IL-5, interferon-gamma (IFN-gamma) in supernatant of the splenocyte cultures and the ratio of IL-4 to IFN-gamma in order to verify the result. RESULTS: Baicalin or icariin at concentrations ranging from 2 to 10 micromol/L, and Astragalus saponin I from 1 to 10 micromol/L effectively suppressed the splenocyte proliferation. When the proportion of baicalin, icariin and Astragalus saponin I was 1:2.14:2.65, the inhibitory effect was most remarkable. Further research confirmed the rationality of the optimal combination. CONCLUSION: An optimal combination of the major effective ingredients from Chinese herbal compound Biminne most effectively suppresses the proliferation of splenocytes from sensitized mice and regulates the cytokine secreting.
Assuntos
Astrágalo/química , Medicamentos de Ervas Chinesas/administração & dosagem , Flavonoides/administração & dosagem , Animais , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Citocinas/metabolismo , Modelos Animais de Doenças , Combinação de Medicamentos , Desenho de Fármacos , Incompatibilidade de Medicamentos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Rinite/imunologia , Baço/citologia , Baço/imunologiaRESUMO
OBJECTIVE: To investigate the effects of Biminne, a traditional Chinese herbal formula, in regulating the endocrine-immune function in mice with airway allergy. METHODS: Forty BALB/c mice were randomly divided into four groups: control group, untreated group, Biminne group, and dexamethasone (Dex) group. The murine model of airway allergy was established by intraperitoneal injection sensitization and intranasal challenge with ovalbumin (OVA). On day 28, the mice were sacrificed and the sera were collected. The lung tissue of mice was removed for haematoxylin-eosin staining and the pathological changes were observed. The spleen cells of mice were isolated and the ratio of Th2/Th1 was determined by flow cytometry. The isolated spleen cells were also cultured in 24-well plates in the presence of concanavalin A and the supernatants were collected after 72-hour culture. Levels of interleukin-4 (IL-4), IL-5 and interferon-gamma (IFN-gamma) in the supernatants and the corticosterone level in sera were analyzed by enzyme-linked immunosorbent assay. RESULTS: Biminne significantly relieved the inflammatory reaction in the lung tissue of airway allergic mice. Compared with the control group, the Th2 cells in the untreated group increased, while the IFN-gamma level decreased and the ratio of Th2/Th1 increased significantly, showing a Th2 response priority. Biminne treatment could down-regulate the Th2 cells and their cytokines IL-4 and IL-5, and up-regulate the Th1 cells and their cytokine IFN-gamma. So the ratio of Th2/Th1 decreased significantly, shifting to the control level. In addition, Biminne also inhibited the reduction of the serum corticosterone of sensitized mice. CONCLUSION: The Chinese herbal formula Biminne can reverse the functional disorder of the endocrine-immune system, which may explain its therapeutic effects on airway allergic disease.
Assuntos
Medicamentos de Ervas Chinesas/farmacologia , Hipersensibilidade/metabolismo , Células Th1/efeitos dos fármacos , Células Th2/efeitos dos fármacos , Animais , Medicamentos de Ervas Chinesas/uso terapêutico , Hipersensibilidade/tratamento farmacológico , Hipersensibilidade/imunologia , Inflamação , Interferon gama/metabolismo , Interleucina-4/metabolismo , Interleucina-5/metabolismo , Pulmão/patologia , Camundongos , Camundongos Endogâmicos BALB C , Fitoterapia , Células Th1/metabolismo , Células Th2/metabolismoRESUMO
OBJECTIVE: To investigate whether the effect of Biminne on allergic rhinitis (AR) was through improving vascular permeability of nasal mucosa. METHODS: Rat's model in Biminne-treated group and model group was induced by intraperitoneal injection of ovalbumin (OVA) and aluminum hydroxide gel suspension Biminne-treated rats were orally given Biminne suspension from the 8th day to the 17th day. On the 18th day, Evan's blue dye (EBD) in the nasal perfusate was detected to assess the vascular permeability. RESULTS: EBD concentration was higher in the model rats than that in the normal rats, and lower in the Biminne-treated rats than that in the model rats (both P < 0.01). CONCLUSION: Biminne could improve vascular permeability of nasal mucosa in sensitized rats, which may be the mechanism of its clinical effect on AR.