| Systemic lupus erythematosus(SLE)was a chronic autoimmune disease and a diffuse connective tissue disease characterized by immune inflammation.It was characterized by the production of autoantibodies and systemic inflammation,which could cause damage to multiple organs,including the kidneys,skin,heart,lungs and central nervous system.The complications were one of the main causes of death in patients with SLE,such as lupus nephritis(LN),neuropsychiatric lupus,and cardiovascular diseases.Lupus nephritis,a serious complication of SLE,showed glomerular mesangial cell hyperplasia in renal pathological biopsy even without clinical symptoms.As the course of the disease progresses,it caused abnormal kidney function and eventually renal failure is one of the leading causes of death in SLE patients.Hormones and immunosuppressants were the cornerstone drugs for the treatment of systemic lupus erythematosus and lupus nephritis.However,the drugs might cause some side effect,such as susceptibility to infection,gastrointestinal reactions,liver and kidney damage,etc.It was important to understand the pathogenesis of LN and find new therapeutic targets.The function of T-cell in patients with systemic lupus erythematosus had been thoroughly studied for decades,and has confirmed defects or changes in their signaling pathways,cytokine production,cell proliferation,or regulatory functions.CD4 + T cells were considered necessary for the development of lupus.Their basic role was to provide auxiliary signals for B cell differentiation and autoantibody secretion,and they were the most effective drivers of B cell differentiation.The recent discovery of TLRs illustrated the central role of naturalimmune system activation in regulating applicable immune responses,inflammation,and tissue repair.It leaded to the activation of members of the IFN regulator family and the activation of members of the NF-κB and MAP kinase family.However,non-TLRs-dependent innate immune activation was caused by molecules such as the cytosolic nucleic acid receptors RIG-1 and MDA5,which enabled different chaperone molecules to trigger signaling through IRFs and NF-κB pathways.Nucleic acid-reactive TLRs and non-TLRs plasmid nucleic acids could lead to the production of IFNs and other types of pro-inflammatory mediators,which also clarified that nucleic acids and nucleic acid-containing immune complexes had a direct pathogenic effect and were involved in disorders of lupus immune.Among them,the activation of TLRs-dependent immune signaling pathways controlled an important immune response in lupus,and TLRs-dependent pathway transfer protein TRAF6 played an important role.Tumor necrosis factor receptor-associated receptor(TRAF6)was a member of the tumor necrosis factor(TNF)receptor related factor protein family,which was widely expressed in mammalian cells,and played an important role in maintaining immune homeostasis,as well as was being involved in inflammation and autoimmunity Of sexually transmitted diseases.As an important adapter,TRAF6 mediated the toll-like receptor signaling pathway and activates the NF-κB signaling pathway by regulating inflammation and immune responses.In addition,it has been reported that TRAF6 is up-regulated in LN patients,which accelerated the progression of end-stage renal disease(ESRD).Revealing the regulatory mechanism of TRAF6 not only providesd theoretical basis for the study of inflammation and infectious diseases,but also brought new opportunities for the treatment of LN.Objective:This experiment investigated the effect of TRAF6 on the proliferation of mesangial cells in lupus nephritis and the expression of peripheral blood cytokines,and performed clinical indicators and related analysis,which provided theoretical basis for the pathogenesis and treatment of lupus nephritis.Methods : 1.Selected outpatient and inpatient patients in the department of rheumatology of Hunan Provincial People’s Hospital from November 2017 to November 2019,collected the clinical data of the patients according to the inclusion and exclusion criteria.The control group was healthy people in the physical examination center of our hospital.Peripheral blood mononuclear cells were collected,total RNA was extracted,and expressions of TRAF6,IL-1β,IL-6,IFN-γ,TNF-α were detected by real-time PCR.2.Randomly collected outpatient and inpatient SLE patients of Hunan Provincial People’s Hospital,the serum was isolated to detect the expression of inflammatory factors and lymphocytes,and the clinical data were analyzed.3.MRL/ LPR mice(30 cases)and normal control group(30 cases)were purchased.Immunohistochemistry was used to detect the expression of TRAF6 in renal tissue of lupus mice.4.The glomerular mesangial cell line was purchased,Establishing a glomerular mesangial cell model of lupus nephritis.Gglomerular mesangial cells were stimulated with different concentrations of LPS,detecting the expression of TRAF6,IL-1β,IL-6,IFN-γ,TNF-α and the effects on the proliferation of glomerular mesangial cells.5.TRAF6 was knocked out to detect the expression of cytokines and their effects on the proliferation of glomerular mesangial cells.All data are from at least three independent experiments.Two independent groups are represented by two-sample t-tests,Multipleindependent groups are represented by one-way analysis of variance(ANOVA),Data are expressed as mean ± standard deviation.p <0.05 was considered statistically significant.All data were analyzed using GraphPad Prism 5 software.Results: 1.Clinical data of SLE patients without LN,LN patients and normal control group:Twenty patients with SLE without LN and twenty patients with LN were collected.The control group selected thirty healthy people from our physical examination center.There was no significant difference in gender and age between the three groups(P>0.05).2.Compared with the healthy control group,TRAF6 mRNA was highly expressed in peripheral blood PBMC of patients with SLE and LN,and higher in LN.3.LN patients had higher expression of inflammatory factors and lower lymphocytes.4.The glomerular mesangial cell model of lupus nephritis was established,and the glomerular mesangial cells were stimulated with different concentrations of LPS,With the increase of LPS concentration,the viability of mesangial cells gradually decreased,and the apoptosis rate increased,TRAF6 mRNA and inflammatory factors had higher expression in LN mesangial cells,which revealed the involvement of TRAF6 and cytokines in systemic lupus erythematosus and lupus nephritis.5.Knockout of TRAF6 reduced the inflammatory damage of HRMCs induced by LPS,and the expression of inflammatory factors was significantly lower than that of the control group,which suggested that TRAF6 was involved in the pathogenesis of LN.6.Immunohistochemistry showed that the expression of TRAF6 protein in kidney tissues of lupus mice(MRL / Ipr)was significantly higher than that of mice(C57BL / 6)。Conclusion:1.TRAF6 may participate in the pathogenesis of LN byaffecting the proliferation of mesangial cells and the secretion of inflammatory factors.2.The high expression of cytokines and the decrease of lymphocytes were one of the important factors in the pathogenesis of systemic lupus erythematosus and lupus nephritis. |
