| Objective:SLE (Systemic Lupus Erythematosus, SLE) is a kind of etiology with diverse clinical symptoms is unknown, involving multiple organ complex autoimmune disease. Its pathogenesis is unclear. Lupus nephritis (lupus nephritis,LN) is the most common clinical manifestations of SLE, accounting for the most common clinical manifestations of 60% ?70%,and its etiology and pathogenesis are still not very clear. Abnormal cytokine expression or cytokine network imbalance has been considered as one of the important factors of the LN.Mesenchymal stem cells (UC-MSC) has been reported as an effective cells in the treatment of LN.Some experiments suggest the expression of osteopontin (OPN) in blood or urine and LN damage degree and the condition changes have a close relationship, which has important diagnostic value and curative effect judgment value.This paper's purpose is to explore the potential therapeutic effect and possible new mechanisms of umbilical cord-derived mesenchymal stem cells (UC-MSC) treatment in lupus nephritis (LN);To investigate the function of osteopontin (OPN) in treatment of LN using UC-MSC and the underlying mechnism.Methods:1. 13 patients with LN, who were type III / IV according to WHO, were assigned to two groups: HUC-MSC transplantation group and placebo treatment group. (solution by the ethics committee)Two or six months after treatment, the protein levels of OPN, AP-1, SP-1,NF-KB, MMP-2 and MMP-9 in serum from the two groups were detected by enzyme linked immunosorbent assay (ELISA).2. (1) Twenty four 22-week-old male MRL/lpr mice were divided into two groups as followings: the group 1 had been treated with 0.5ml PBS as control, and the group 2 had been transplanted with HUC-MSC labelled by SPIO-PLL (super paramagnetic iron oxide polylysine) . The distribution of HUC-MSC in mice was detected by MRI. (2) The bodyweight were assessed every two weeks. The levels of urine and relevant immunological parameters were determained by ELISA. (3) HUC-MSC had been transfected with lentivirus carrying GFP (green fluorescent protein ) . MRL/lpr mice were transplanted with GFP-HUC-MSC through caudal vein. The expressions of OPN in the kindey were detected.3. Twenty four 22-week-old male MRL/lpr mice were divided into two groups as following:the group 1 had been treated with 0.5ml PBS as control, and the group 2 had been transplanted with 1×106 P3 HUC-MSC. Another group was including twelve 22-week-old male C57BL/6 mice as the negative control. The mRNA expressions of OPN,AP-1,SP-1,NF-?B, MMP-2 and MMP-9 in renal tissues were determained by Q-PCR. The protein expressions of OPN, AP-1, SP-1, NF-?B, MMP-2 and MMP-9 in kindney were determained by western blot and immunohistochemistry.Results:1.The expression levels of OPN, MMP-2 and MMP-9 in serum of LN 'patients were significantly down-regulated in renal tissues from Huc-MSC-transplantated patients compared with that from the control. All have significant difference (P < 0.05)2.The expression levels of AP-1,SP-1,and NF-?B in serum of LN patients were down-regulated in renal tissues from Huc-MSC-transplantated patients compared with that from the control.3.In Huc-MSC transplantation group, proteinuria concentrations, serum ds-DNA titer and serum ANA titer were down-regulated compared with the control group. Body weight of mice decreased obviously4.The levels of OPN, MMP-2 and MMP-9 in serum of MRL/lpr mice, were significantly down-regulated from Huc-MSC transplantation group compared with that from the control mice. (P < 0.05)5.Both of mRNA expression and protein expression levels of transcription factors AP-1,SP-land NF-?B were significantly downregulated in renal tissues from Huc-MSC-transplantated mice compared to that from the control group. (P < 0.05)6. By immunohistochemical staining, the positive staining of transcription factor AP-1, SP-1 and NF-?B were much weaker in renal tissues from Huc-MSC-transplantated mice than the control renal tissues. The expression levels of OPN, MMP-2 and MMP-9 were also significant lower in renal tissues from Huc-MSC-transplantated mice than that from the control group.Conclusion:HUC - MSC transplantation can effectively reduce the OPN in serum of LN patients, AP - 1,SP - 1, /P65, the expression of MMP - 2 and MMP - 9 levels, the treatment of LN patients may have a certain role;HUC - MSC transplantation treatment called simply MRL/LPR mice have certain curative effect with lupus nephritis efficacy: transplantation treatment group serum decreased significantly, the expression of OPN content in serum of transcription factors,SP - 1, AP - 1 MMP - 2 and MMP - 9 and NF-?B expression level in protein and RNA and lupus mice in the glomerular mesangial cells and renal tubular expression significantly reduced .The results provide a new mechanism that treatment of Huc-MSC in MRL/lpr mice mainly inhibited the expression of OPN through reducing AP-1 and SP-1 expressions. The down-regulation of OPN subsequently induced the reduction of downstream molecules MMP-2 and MMP-9 via NF-1aB pathway. These findings will aid our understanding of the treatment of SLE with MSC. |
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