RESUMO
OBJECTIVE: To establish an HPLC method for simultaneous determination of four kinds of purines in deer fetus soft capsule. METHODS: Four kinds of purines were detected and determined by HPLC. The mobile phase was 0.02 mol/L KH2PO4 (containing 1 mmol/L heptane sulfonic acid sodium, pH = 3.8)-methanol = 97:3. Detection wavelength was 254 nm and flow rate was 1.5 ml min. The linear relationship of four kinds of purines was as follows: hypoxanthine: Y1 = 83695X1 + 355 (r1 = 0.9998), with the linear range 0.040-0.667 mg/mL; xanthine: Y2 = 50638X2 + 39 (r2 = 0.9989), with the linear range 0.008-0.119 mg/mL; guanine: Y3 = 30269X3-9562 (r3 = 0.9924), with the linear range 0.018 - 0.279 mg/mL; adenine: Y4 = 38975X4-8671 (r4 = 0.9989), with the linear range 0.027-0.399 mg/mL The average sample recovery rate of hypoxanthine, xanthine, guanine and adenine were 98.1%, 98.6%, 98.0% and 97.5%, with RSD 1.0%, 0.4%, 0.8% and 0.6%, respectively. RESULTS: The content of hypoxanthine, xanthine, guanine and adenine in 3 lots of deer fetus soft capsule were 116.5-132.0 microg/capsule, 21.2-23.0 microg/capsule, 48.6-54.3 microg/capsule and 68.9-75.2 microg/capsule, respectively. CONCLUSIONS: This method is simple,accurate and reproducible, which provides a basis for quality control of purines in deer fetus soft capsule.
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Cervos , Materia Medica/química , Purinas/análise , Adenina/análise , Animais , Cápsulas , Feto , Guanina/análise , Hipoxantina/análise , Controle de Qualidade , Reprodutibilidade dos Testes , Xantina/análiseRESUMO
This study was aimed to explore the effect of cordyceps sinensis enhancing lymphocyte proliferation and surface CD marker expression in simulated microgravity environment. The splenic lymphocytes were separated from mice and cultured in the rotary cell culture system simulated microgravity environment. The cells were treated with different concentration of cordyceps sinensis solution (0, 6.25, 12.5, 25 and 50 µg/ml) for 24, 48 and 72 h respectively, then the cells were harvested, and analyzed for cell proliferation and the expression of cell surface markers (CD4 and CD8). The results showed that under simulated microgravity environment, the lymphocyte proliferation was inhibited. When the concentration of cordyceps sinensis was 25 or 50 µg/ml, the lymphocyte proliferation, CD4 and CD8 expressions all increased, but 50 µg/ml cordyceps sinensis could inhibit the proliferation ability with the time prolonging. It is concluded that the suitable concentration of cordyceps sinensis displayed the ability to enhance the lymphocyte proliferation and CD marker expression in simulated microgravity environment. These results may be valuable for screening drugs which can be potentially against immunosuppression under simulated microgravity.
Assuntos
Cordyceps , Linfócitos/efeitos dos fármacos , Polissacarídeos/farmacologia , Simulação de Ausência de Peso , Animais , Antígenos CD4/metabolismo , Antígenos CD8/metabolismo , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Tolerância Imunológica , Ativação Linfocitária/efeitos dos fármacos , Linfócitos/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Baço/citologiaRESUMO
This study was aimed to evaluate the effect of lentinan on the immune function of splenic lymphocytes in rotary cell culture system (RCCS) microgravity environment. The splenic lymphocytes from mice were separated and cultured in the normal gravity and the microgravity environments. The cells were treated with lentinan solution (0, 10, 20 and 40 µg/ml). After incubated with lentinan for indicated times (24, 48 and 72 h), the cell proliferation, secretion of cytokine and the expression of cell surface markers were detected by MTT method, ELISA and flow cytometry respectively. The results indicated that lentinan of above mentioned concentrations did not obviously promote the lymphocyte proliferation, but increased the secretion of IL-2 and IFN-γ and enhanced the expression of lymphocyte surface markers CD4 and CD8 in microgravity environment. It is concluded that lentinan has the ability to enhance the lymphocyte immune function in microgravity environment.