| Backgrounds and ObjectivesSystemic lupus erythematosus (SLE) is an autoimmune syndrome, and involves various organs, in which kidney is mostly common involved. Lupus nephritis (LN) is a common secondary glomerular disease with diverse clinical manifestations and pathology. Renal involvement is an independent risk factor associated with the poor prognosis of SLE, and the renal failure is the main dead cause of SLE. In recent years, the survival rate in LN has significantly increased by applying glucocorticoid and first-line immunosuppressive agents. However, not all patients benefited, approximately 35% patients relapsed, and 10% ~ 20% patients progressed to end-stage renal disease during 10 years, moreover, many of them died of adverse reactions caused by long-term use of immunosuppressive agents. Therefore, new and better tolerated immunosuppressive or biological agents are urgently needed. Leflunomide (LEF) is a new class of immunosuppressive agents, which is a kind of anti-proliferative activity of isoxazole. Recently, LEF has mainly been used for the clinical treatment of LN. Because of few sample in the clinical research, so the exact mechanism needs further study.A chronic graft-s-host disease (cGVHD) lupus mouse model was established to research the efficacy of Leflunomide (LEF) on serum ANA, dsDNA antibodies, IgG and IgM immunity fluorescence in kidney, pathological changes of kidney, spleen cells proliferation, TLR9 mRNA expression in spleen in order to investigate the effect and partial mechanism of Leflunomide on lupus nephritis mice.MethodsMice were divided into three groups: model group (LN mice induced by cGVHD, n=12) mice, LEF(LN mice were treated by LEF,n=10) mice and control (n=12) mice. Model group was established through cGVHD induction after 11 weeks. Then 20 mg/kg·d dosage of LEF was lavaged for LEF group; similarly, the same dosage of physiological saline was supplied to control and model groups. 10 weeks later, blood, kidney and spleen were collected from the three groups. Detecting the titres of serum ANA, dsDNA antibodies. IgG and IgM in kidney were observed under microscope. Check pathological changes through florescent labeling and PAS, PASM staining, respectively. Reverse transcription-polymerase chain reaction for mRNA expression. The spleen cells from control group were cultured and reproduced on 96-well plates and were divided into four groups (medium group, C274 group, LEF+C274 group and LEF group). The OD values were monitored by MTT enzyme detector in 570nm to detect the inhibition of LEF on cell proliferation.Results(1) Serum ANA and dsDNA antibodies concentrations in LEF group significantly decreased comparing to model group (p<0.05). (2) LEF group was significantly different with model group with less sediment of IgM and IgG in kidney and AI declined dramatically, and pathological changes mainly were based on cell membranes and matrix slightly proliferation or diffusely heavy proliferation (p<0.05). (3) TLR9 mRNA expression significantly decreased in LEF group compared with model group (p<0.05). (4) Spleen SI values in LEF+C274 group were significantly decreased compared to C274 group (p<0.05). ConclusionsLEF significantly downregulated the titres of serum ANA, dsDNA antibody, and could restrain the deposition of IC in the kidneys, reduce the AI index, reverse kidney pathological changes, effectively restrain the spleen cells proliferation and TLR9 mRNA expression. LEF significantly inhibit the progression of lupus nephritis mice induced by cGVHD.
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