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AIM: To investigate the effect of berberine (Ber) on the activation and proliferation of T lymphocytes and its mechanism of action. METHODS: Whole peripheral blood from normal subjects was stimulated with phytohemagglutinin (PHA) or phorbol ester (PDB) plus ionomycin (Ion) and the expression levels of CD69 and CD25 were evaluated with flow cytometry after the staining with appropriate fluorescent monoclonal antibody. The distribution of cell cycles was analyzed by propidium iodide staining and dead cells by 7-aminoactinomycin live staining. RESULTS: 100 ?mol/L and 50 ?mol/L of Ber had significant inhibition of the expression of CD69 on T cells stimulated with PDB plus Ion or PHA, while effect of 25 ?mol/L Ber was not significant. And as time of action extended, the extent of inhibition decreased. For the expression of CD25, Ber at the concentrations as above all exerted significant inhibitory effect in a dose-dependent manner. Moreover, Ber could block lymphocytes cell cycle progression from G_0/G_1 phase to S and G_2/M phase without phase specificity. Besides, live staining analysis revealed that Ber did not have significant cytotoxicity on lymphocytes. CONCLUSIONS: Ber significantly inhibits the expression of early and mid activation antigens of T cells and also blocks the progression of lymphocytes cell cycles. These results suggest that Ber exerts immunosuppression effect through inhibiting the activation and proliferation of T cells.
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Object To look for the proprietary constituent and the constituents with blood lipid regulating effect from the dried fruit of Crataegus pinnatifida Bge var major N E Br Methods Various column chromatographic techniques were employed for isolation and purification of the constituents UV, IR, EI MS, FAB MS, 1HNMR, 13 CNMR, HMBC, HMQC and 13 CGATE were used to identify the structure of the isolated constituents Results Six compounds were isolated from the fruit of C pinnatifida They were identified as 5, 7, 4′ trihydroxy flavone 8 C ? L rhamnopyranosyl (1→2) ? D glucopyranoside, i e vitexin rhamnoside (Ⅰ), hyperoside (Ⅱ), citric acid (Ⅲ), vitexin (Ⅳ), quercetin (Ⅴ), ursolic acid (Ⅵ) Conclusion Compound Ⅰ is a proprietary constituent in the fruit of C pinnalifida, which was obtained from this plant for the first time Compound Ⅱ is a main constituent with blood lipid reducing effect in flavonoids
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AIM: To investigate the characterization of absorption of hematoporphyrin monomerthyl ether (HMME), a domestic new generation photosensitizer product, by activated T cells from human peripheral blood. METHODS: Evaluation was performed by flow cytometry on the effects of incubating concentration and time of HMME on absorption by activated T cells. Lymphocytes were separated from human peripheral blood by density gradient centrifugation with Ficoll and T cells were activated with polyclonal stimulators PHA and PDB+Ion. To analyze the effects of HMME incubating doses on the absorption of activated T cells, the cultural lymphocytes were incubated with a serial doses of HMME for 1 h and HMME absorption were measured by FACS after immuno-staining with anti-CD3 antibody. To test the impact of HMME incubating time on the absorption of activated T cells, the cultural lymphocytes were incubated with HMME for various times and HMME absorption were measured by FACS after immuno-staining with anti-CD3 antibody. RESULTS: The HMME absorption-dose curve and absorption-time curve were shifted to right and up in the activated T cells as compared to resting T cells. HMME absorptions of activated T cells were statistic significantly larger than that of resting T cells in the doses between 5 mg/L to 20 mg/L. HMME absorptions of either activated T cells or resting T cells underwent a gradual increase with the incubation-time in HMME at concentration of 10 mg/L. HMME absorptions of activated T cells were statistic significantly larger than that of resting T cells in the incubation-time between 15 to 60 min. CONCLUSION: The differences of HMME absorption between activated T cells and resting T cells depend on the incubation times and doses of HMME. HMME absorption of activated T cells are significantly larger than that of resting T cells in certain incubation-times and doses. These results suggest that incubation time and dose associated with HMME-PDT therapeutic windows will be created for selective deletion of activated T cells.
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AIM: To investigate the inhibitory effect of quercetin on in vitro activation of T lymphocytes by polyclonal activators with CD69 expression as an activation marker. METHODS: After being separated from lymphoid nodes of a C57BL/6 mouse, the lymphocytes were exposed to polyclonal activators (PDB or Con A) with or without quercetin. Then they were harvested at 2 h, 6 h and 24 h, respectively. The expressional rates of CD69 on T lymphocytes were assessed by two-color immunofluorescent staining and flow cytometry, and the inhibitory rates of quercetin at different time points were estimated. RESULTS: Quercetin had no effect on the expressional rate of CD69 on T lymphocytes under resting states. After the stimulation with PDB or Con A, the expressional rates of CD69 on T lymphocytes in the present of quercetin (10 ?mol/L) showed significant decrease compared with those of control groups at different time points (P
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AIM: The expression of CD28, CD56 and CD57 on CD8+T cells in the peripheral blood of young (age range 20-35) and elderly(age range 60-75) healthy donors were compared to explore the change of the cellular immune function with aging.METHODS: Three-color fluorescent flow cytometry was performed to analyze the differences in percentage of CD8+CD28+, CD8+CD56+ and CD8+CD57+T cells in the peripheral blood between the young and elderly groups.RESULTS: CD8+CD28+T cells in the peripheral blood of the elderly group was significantly lower than those in the young group, with percentage of 34.07?5.28 and 49.84?7.43,respectively (P