Experimental Study On Eczema Nanocream Composition On The Lymphocytes And Hacat Cell Co-culture

Background and Objective:Chronic eczema is a common clinical disease, frequently-occurring disease, its etiology is complex, a variety of skin lesions, recurrent, severe itching, the treatment of difficult, seriously affect the patient’s physical and mental health. The results confirmed that the cytokine induced by chronic eczema patients with the differentiation of Thl∨Th2cells imbalance secretion affects the development of eczema, the skin lesions of patients with chronic eczema hints of infiltration of T lymphocytes play an important role in the pathogenesis of chronic eczema, forming cell keratinocytes induced activation of T cells apoptosis as the turning point of eczema chronicity, blocking occurs horny the formation of cell apoptosis can reduce chronic eczema process. HaCaT cell lines maintained normal human keratinocyte differentiation and proliferation characteristics of cells, formation of cell physiological function of horny generic model, exploring the use of lymphocytes and HaCaT cells were cultured in vitro cell model to establish chronic eczema, efficacy for chronic eczema experiment and drug screening test has important significance.Eczema nano cream for Professor Yang Zhibo based on years of clinical experience in the treatment of chronic eczema, external preparation of the combination of nanotechnology, preclinical studies demonstrate efficacy. This study intends to explore the lymphocytes and HaCaT cells, apoptosis of eczema nano cream composition on chronic eczema in vitro cell model of HaCaT cells was established and eczema, mechanism of nano cream constituents of chronic eczema cell model in vitro for the treatment of chronic eczema, provide the basis for establishing the in vitro cell model of chronic eczema and the clinical medication.Method:Through the orthogonal design experiments to explore the best model to establish apoptosis of lymphocytes and HaCaT cells co-culture of pathogenesis of chronic eczema, simulation of cells in vitro, concentrations of MTT are used to study the eczema nano cream main component of the cell viability of HaCaT, drug intervention in cells was detected by flow cytometry and TUNEL co-culture apoptosis rate of HaCaT cells model.Results:Chronic lymphocyte in patients with eczema and HaCaT cells to1:1,1:3,3:1ratio were inoculated separately co-culture of24h,48h,72h, lymphocytes could induce apoptosis of HaCaT cells; Lymphocytes and HaCaT cells in a co-culture of72h, inoculation ratio of Lymphocyte:HaCaT=3:1is the best co-culture apoptosis model; Mat in50ug/mL and concentration on the growth of HaCaT cells had no obvious inhibitory effect, the concentration of12.5ug/mL can promote the apoptosis of co-cultured model, the concentration of25ug/mL reversal effect is not apparent, the concentration of50ug/mL reversal effect. Oxy in50ug/mL and HaCaT cell activity had no effect on the concentration,12.5ug/mL concentration had a stimulating effect on co-culture apoptosis model,25ug/mL,50ug/mL levels were reversal effect. Promote, reversal effect of Oxy on apoptosis in the co-culture model was more strong than Mat; Ber in the concentration of1.5mg/L on the proliferation of HaCaT cells had no obvious effect, Ber has the reverse effect on the co-culture apoptosis model was dose dependent and Ber concentration; No significant influence of concentration of Pae in the15.625mg/L proliferation of HaCaT cells,75mg/L,15mg/L had no effect on the apoptosis of co-cultured model,7.5mg/L concentration had a stimulating effect on co-culture apoptosis model; Bor in the concentration of125umol/L on the proliferation of HaCaT cells had no obvious effect, Bor has the reverse effect on the co-culture apoptosis model was dose dependent and Bor concentration; Men in the concentration of9.38mg/L on the proliferation of HaCaT cells had no obvious effect, Men has the reverse effect on the co-culture apoptosis model was dose dependent and Men concentration.Conclusion:1.Eczema patients with chronic lymphocytes co-cultured with HaCaT cells, lymphocytes can induce the apoptosis of HaCaT cells, Lymphocyte:HaCaT=3:1is the best inoculation proportion co-culture apoptosis model.2.Mat can promote the apoptosis of co-culture model in12.5ug/mL concentration,50ug/mL concentration reversal effect. Oxy can promote the apoptosis of co-cultured model when the concentration of12.5ug/mL,25ug/mL,50ug/mL levels were reversal effect. Promote, reversal effect of Oxy on apoptosis in the co-culture model was more strong than Mat.3.Ber in the concentration of1.5mg/L on the proliferation of HaCaT cells had no obvious effect, Ber has the reverse effect on the co-culture apoptosis model was dose dependent and Ber concentration.4.Pae in the concentration of15.625mg/L on the proliferation of HaCaT cells had no obvious effect,75mg/L,15mg/L had no effect on apoptosis model of co-culture,7.5mg/L concentration had a stimulating effect on co-culture apoptosis model.5.Bor in the concentration of125umol/L on the proliferation of HaCaT cells had no obvious effect, Bor has the reverse effect on the co-culture apoptosis model was dose dependent and Bor concentration.6.Men in the concentration of9.38mg/L on the proliferation of HaCaT cells had no obvious effect, Men has the reverse effect on the co-culture apoptosis model was dose dependent and Men concentration.