| Objective: To explore whether ozonated water can improve the symptoms of the mouse model of allergic contact dermatitis(ACD)induced by dinitrochlorobenzene(DNCB)as well as its possible mechanism,and to afford theoretical foundations for new methods to treat allergic contact dermatitis,thus providing a more reliable basis for further clinical application.Methods: Sixty BALB/c mice were randomly divided into two groups: 50 mices for the experimental group,10 mices for the control group.Applied DNCB on the mices for the ACD model,and applied the same amount of acetone and oil on the mices in the control group,then executed the mices which randomly selected 10 mices from the experimental group and all of the control group after establishing the model,finally confirmed successful establishment of the mouse ACD model through the detection of the serum and pathology of tissues and electron microscopy and cell factors related to skin lesion.The left mice in experimental group were averagely divided into four groups at random: ozonated water group,purified water group,positive medication control group and spontaneously-recovered model group.Two weeks later,we got the whole skin lesions and blood sample from the mices after execyted them,then observed the pathology of skin tissues of each group with HE dyeing;detected the level of IgE and IL-4 in blood serum by ELISA;The changes of skin spines in mice were observed by electron microscopy;and the level of IFN-γ,IL-10 and IL-17 in the skin lesions were quantitatively detected by PCR(RT-PCR).Results:1)The manifestation such as erythema,anabrosis,obvious scabs and exudation showed up on the back of all mices in the model group which showed the typical clinical symptoms of allergic contact dermatitis;only skin conditions of mices with acetone and oil application from the control group showed no overt difference from the beginning,as the level of these cells factors and the pathology were in line with reports of relevant documents and papers,the mouse ACD model had been successfully established.2)In two weeks after difference treatment to each group,it was observed that horny layers of the ozonated water group and the medication group were significantly thinner than that of the model group;the thickness of epidermis of the former two groups got normal while no overt change happened to the purified water group and the spontaneously-recovered model group.Chromatin in spinous layer was no aggregation by the electron microscope,mitochondrial morphology structural integrity and no swelling,and the ridge line was clear in the ozonated water group.The expression of mitochondria in the medication group was complete,no rupture,swelling,vacuolization and prominent nucleoli were seen.Nucleolus of Purified water treatment group was not obvious,mitochondria vacuolization and ridge disappear.The Nucleolus of the spontaneously-recovered model group were irregular,and the vacuoles of the mitochondria were serious,the ridge disappeared and accompanied slight swelling.3)Besides,the levels of IgE in blood serum detected via ELISA in the ozonated water group and the medication group obviously dropped down compared to that of the spontaneously-recovered model group,statistical significance manifested itself for p<0.05(t=10.641、9.839,p=0.000、0.000,p<0.01).The levels of IL-4 in blood serum detected via ELISA in the ozonated water group and the medication group obviously dropped down compared to that of the spontaneously-recovered model group,statistical significance manifested itself for p<0.05(t=6.873、7.448,p=0.000、0.000,p<0.01).4)The expression levels of IL-10 detected by RT-PCR in skin tissues of the ozonated water group was clearly lower than that of the spontaneously-recovered model group(t=-4.688,p=0.000,P<0.05).The expression levels of IL-17 detected by RT-PCR in skin tissues of the ozonated water group was clearly lower than that of the spontaneously-recovered model group(t=-6.282,p=0.025,P<0.05).The level of IFN-γ for the ozonated water group higher than that of the spontaneously-recovered model group,statistical significance manifested itself for p<0.05(t=-6.282,p=0.025,p<0.05);Conclusion:1.DNCB-sensitized BALB/c mouse model shared similarity with symptoms of human’s allergic contact dermatitis;2.Observed by electron microscop that ozone therapy can promote skin recovery;3.Ozone could be adjusting and controlling the imbalance of T cells and cell redox reaction to reach a therapeutic function. |
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