Research On Relativity And Level Of IL-12, IFN-γ, NO, INOS In Blood Serum Of Onychomycosis Patients

Objective: As a frequently-occurring disease in dermatology department, onychomycosis is a chronic and infectious disease caused by pathomycete invading nail or toenail. It has an increasing tendency year by year due to all kinds of well known reasons.Trichophyton rubrum is the most common dermatophyte causing the fungal skin infections in humans, and it distributed all over the world, and also a common pathogenic mycetes in China. It is an imperatively solve problem in clinic because onychomycosis caused by this fungi is extremely difficult to cure and reinfects easily, which causes patients'quality of life decreased.With the development of study on antifungal immunity, many studies reported that the innate immunity can directly kill the pathogen and generate the acquired immunity. It is the first step of arousing host to generate immune response that the pathogen of invading mycosis is identified by the natural immunity system. Once infected by fungus, phagocytes in body will be activated and excrete many kinds of cell factors and mediators of inflammation as interleukin12 (IL-12). IL-12 is a key factor in generating the natural immunity response, it is necessary that IL-12 activates Th1 cell mediated immune respose.IL-12 plays an important role in natural immunity and acquired immunity. As a pro-inflammatory cytokine, IL-12 is also the bridge between those two immune reponses. Moreover IL-12 can forcefully induce NK cell and APC activation in the natural immunity and acquired immunity Th1 cell and so on, then some cytokines as IFN-γare secreted by these activated immune cells.The generation of IFN-γrelies on IL-12, meanwhile it is also an effective inducing factor of IL-12, and can increase the secretion of IL-12. IFN-γcan induce the macrophages to produce inducible NO synthase (iNOS), and then to generate NO, an important effector molecule clearing pathogen. The factors above-mentioned form a regenerative feedback pathway, which can accelerate immunity response type I as a functional unit and becomes a nexus liking natural immunity and acquired immunity. At present, it is not completely clear about the specific immune nosogenesis of onychomycosis. Being an extended structure of horny layer of organism, nail is also regulated by organism natural immune and acquired immune. The previous researches proved that many cells with immune function and cytokines secreted by them play an important role in the cutaneous mycotic infection, for example Langerhans cell, keratinocyte, capillary blood vessel endothelial cell,Thl and Th2 cells.As a result, the mycotic infection will present the different development courses due to the participation of these cells.Some researches indicate that the property of immune response in human body to trichophyta antigen has a correlation with the severity of dermatophytosis.Trir2,trichophytom rubrum allergen, can induce the derma to produce immediate hypersensitivity (IH) and delayed type hypersensitivity (DTH). IH is related to the chronic and persistent infection characterized by mild inflammation and appearance of IgE antibody, and DTH is related to the severe inflammation reaction and the endurance to reinfection, it indicates that the cell immunity is more effective in relieving dermatophyte infection and building up the protective mechanism. The recent researches on the immune function of onychomycosis patients found that the abnormal changes existed in sub-group of T cell in peripheral blood, including the decrease of CD3+, D4+ and CD4+/CD8+, and the increase of CD8+. These results implies that an unbalanced immune system exists in onychomycosis patients, and the cell immunity presents some extent of effect on occurrence of this kind of disease. In this study we detect the level of IL-12, IFN-γ, NO and iNOS in blood serum of onychomycosis patients, to investigate their effect on onychomycosis nosogenesis, and further to understand the correlation between the natural immune condition and the susceptibility to mycetes in human body, and the effect of specific immune on resisting dermatophyte.Methods:The case group involved thirty five typical onychomycosis patients who were selected in outpatient clinic in the second hospital of Hebei Medical University. Case Group (onychomycosis patients group) Selecting Standards:①All the patients should have typical onychomycosis clinical manifestation. Fungus microscopic examination is positive and Trichophyton rubrum had been cultured.②The age of all the patients in study were between 18 and 55 and women are not pregnant and lactation.③All the cases involved had not taken antimycotic drug for one week, or taken antifungal agent for one month, or drugs that could affect immunological function for two months such as corticosteroids and immunosuppressive agent.④Obtaining the patients'permission. Excluding standards: patients with nail impairment such as psoriasis,chronic eczema,onychodystrophy, etc. And patients with autoimmunity diseases, tumor, acute infection or other severe Systemic diseases should be excluded. The control group involved twenty healthy people without dermatophyte infection, autoimmunity diseases and family history. There is no significant diffidence in the age and sex between case group and control group by t test. Their blood (5ml) were collected and separated and stored at -20℃in both groups. Serum levels of IL-12 and IFN-γwere detected by ELISA. Serum levels of iNOS and NO were detected by spectrophotometry. Specific operation is strictly in accordance with the directions in the test kit. Experimental data adopted SPSS13.0 statistics software, two samples t detection and linear correlation analysis.Results:1. IL-12 serum concentrations were lower in onychomycosis patients than in controls.There is no significant deviation between case group and control group(P>0.05).2. There was lower expression of IFN-γin onychomycosis patients than in controls.There is significant deviation between case group and control group (P<0.01).3. The serum concentrations of NO were lower in onychom- ycosis patients than in controls. There is significant deviation between case group and control group(P<0.01).4. There was lower expression of iNOS in onychomycosis patients than in controls. There is significant deviation between case group and control group (P<0.01).5. In onychomycosis,there is significant correlation between the serum levels of NO and iNOS (r =0.811,P<0.01).Conclusion:1.The serum level of IL-12, IFN-γ, NO and iNOS in onychomycosis patients from whom Trichophyton rubrum has been isolated is lower.2.The immune function of in onychomycosis patients from whom Trichophyton rubrum has been isolated is lower. Maybe it is one of the causes resulting in the repeated infection and the protracted course of disease.