| Objective:On the basis of previous studies, this study explore that the active ingredients of cortex dictamni dictamnine intervene that peripheral blood lymphocytes from patients with chronic eczema induce apoptosis in HaCaT cells.objectively evaluate dictamnine function mechanism of the treatment of eczema, and provides theoretical and experimental basis for further research.Methods:Using MTT for detection of different concentrations and different time dictamnine influence on proliferation of HaCaT cells.Employ Human lymphocyte isolation fluid and separate peripheral blood lymphocytes from patients with chronic Eczema, with density gradient centrifugation.Co-culture of lymphocytes with HaCaT cells with3:172h induced apoptosis of HaCaT cells,Dictamnine intervene co-culture model without affection the viability of HaCaT cells,with Annexin V/PI double staining and TUNEL method to detect apoptosis rate of HaCaT cells.Results:MTT test results show that compared with the normal group dictamnine significantly inhibite the proliferation of HaCaT cells in23ug/ml concentrations, and along with the increase of drug concentration the effect is significantly enhance, has a dose dependent.Dictamnine is no clear time-effect relationship for the proliferation of HaCaT cells, but72h has the strongest inhibition.Under the concentration of11.5ug/mL,dictamnine is no obvious inhibition for the growth of HaCaT cells. The tests show that with Annexin V/PI double staining and TUNEL method to detect apoptosis,Dictamnine has a reverse effect on apoptosis in HaCaT cells;with the increase of concentration, the reverse rate increased significantly.the result prompt that the reversal effect has a dose dependent with dictamnine.Conclusion:The peripheral blood lymphocytes of Chronic eczema patients can cause HaCaT cells apoptosis.Under the concentration which don’t affect the viability of HaCaT cells, Dictamnine has a reverse effect on apoptosis in HaCaT cells and has a dose-dependent with dictamnine. |
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