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Systemic lupus erythematosus (SLE) is a chronic autoimmune disease characterized by multi-organ and multisystemic involvement and various clinical manifestations.Bone marrow mesenchymal stem cells (BM-MSCs),a kind of non-hematopoietic stem cells originating from the mesoderm,are key components of hematopoietic microenvironment.Recent studies have indicated that SLE is a disorder of stem cells.Both hematopoietic and mesenchymal stem cells (MSCs) are abnormal in SLE,which mainly manifests as changes of biological characteristics,abnormal cytoskeleton and ultrastructure,shortened telomeres,increased telomerase and SA-β-Gal acitivity,decreased differentiative ability,aberrant immunoregulatory effect,and other features of senescence.The mechanism of MSC aging may be related to up-regulated expressions of aging-related genes p53/p21cip1,p16INK4a/Rb and p27kip1/pTEN,elevated levels of reactive oxygen species (ROS),endoplasmic reticulum stress,epigenetic alterations,etc.
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Objective To evaluate biological behaviors of bone marrow mesenchymal stem cells (BMSCs) from patients with systemic lupus erythematosus (SLE),to confirm that the BMSCs are aging stems cells,and to explore mechanisms underlying their aging.Methods BMSCs were isolated from bone marrow of 6 patients with SLE (patient group) and 8 healthy human controls (control group) by density-gradient centrifugation and plastic adherence,and cultured in vitro.Optical microscopy was conducted to observe morphological changes and growth of BMSCs,and growth curves were drawn.The differentiation ability of BMSCs was evaluated through culture of them with adipogenic and osteogenic induction medium.Flow cytometry was performed to identify cellular surface markers and to analyze cell cycle and apoptosis,and scratch assay to assess the migration ability of BMSCs.Immunofluorcscence assay and Western blot analysis were carried out to analyze the distribution and expression of p27kip1/PTEN in BMSCs respectively.Results Patient-derived BMSCs,which had a broad,fiat and polygonal shape,showed decreased growth rate,migration activity as well as adipogenic and osteogenic ability compared with those from the controls.There was a significant increase in the proportion of BMSCs in early stage (17.98％ ± 3.26％ vs.8.23％ ± 3.25％,t =3.91,P =0.011) as well as in middle to late stages (16.80％ ± 9.63％ vs.3.33％ ± 2.21％,t =2.99,P=0.048) of apoptosis in the patient group compared with the control group.Moreover,compared with the control group,the patient group showed a significantly higher proportion of BMSCs arrested in the G0/G1 phase (92.34％ ± 5.80％ vs.78.65％ ± 3.22％,t =3.635,P =0.015),but a lower proportion in the S phase (0.86％ ± 1.72％ vs.5.06％ ± 1.874％,t =3.084,P=0.027).The protein expressions of p27 and PTEN were significantly higher in the patient group than in the control group (p27/β-actin:t =2.784,P=0.039;PTEN/β-actin:t=4.812,P =0.041).Conclusion The BMSCs from SLE patients exhibit senescence-related features,which may be associated with elevated expression levels of p27kip1/PTEN.
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Objective To detect the expression of Blimp1 gene in MRL/lpr mice,so as to provide new ideas for plasma cell-targeting therapy of systemic lupus erythematosus (SLE).Methods Electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation analysis (ChIP) were performed to evaluate the regulatory effect of Blimp1 on B cell maturation antigen (BCMA),and fluorescence-based quantitative PCR was carried out to detect the expression of Blimp1 mRNA in spleen and lymph node tissue of MRL/lpr mice and normal control mice.The intergroup difference in Blimp l mRNA expression was assessed by rank sum test.Results Blimp1 could bind to the BCMA gene promoter.Increased Blimp1 mRNA expression was observed in both the spleen and lymph node tissue of MRL/lpr mice compared with the normal control mice (Z =2.609,3.402,respectively,both P ＜ 0.01).Conclusions BCMA appears to be the target gene of Blimp1,and Blimp1 may plays a certain role in the maintenance of plasma cell survival and antibody production via directly regulating BCMA gene expression.
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ObjectiveTo investigate the action mechanism of Th17 cells in immunoinflammatory response in systemic lupus erythematosus(SLE).MethodsReverse-transcription PCR was performed to measure the mRNA expre ssion of the retinoic acid receptor-related orphan nuclear receptor RORγt in peripheral blood mononuclear cells (PBMCs) from 12 patients with active SLE,9 patients with inactive SLE and 12 normal human controls.Data were statistically analyzed by approximate F test(Welch test) and Dunnett's T3 multiple comparison test (corrected).ResultsIn the case of RORγt mRNA expression in PBMCs,significant differences existed among the 3 groups(F =23.286,P ＜ 0.01 ); in detail,the patients with active SLE were significantly higher than patients with inactive SLE and normal controls ( 1.06 ± 0.44 vs.0.65 ± 0.25,F =2.453,P ＜ 0.05;1.06 ± 0.44 vs.0.22 ± 0.08,F =6.504,P ＜ 0.05),and the patients with inactive SLE were significantly increased compared with the normal controls (F =3.343,P ＜ 0.05).The expression level of RORγt mRN A was significantly positively correlated with SLE disease activity index (rp =0.623,P ＜ 0.01 ).ConclusionsThere is a polarization of Th17 cells in patients with SLE.To antagonize the transcription factor RORγt,which plays an essential role in the regulation of Th17 cell differentiation,may facilitate the control of SLE via attenuating the immunoinflammatory response.
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Objective To study the expression of interferon regulatory factor 1 (IRF-1) in patients with systemic lupus erythematosus (SLE) and the influence of prolactin (PRL) upon it. Methods The level of serum PRL in quiescent condition was examined by electrochemiluminescence-meter in 30 patients with SLE and 20 healthy controls. Peripheral blood mononuclear cells (PBMC) were separated from all the subjects by gradient centrifugation density, and cultured with or without the presence of recombinant human PRL (rhPRL) for 24 hours. The expression of IRF-1 gene in cultured PBMC was detected by reverse transcriptase-PCR (RT-PCR) with gel image scanning. Results The relative value of IRF-1 gene expression was significantly higher in SLE patients than in normal controls (0.89±0.21 vs 0.78±0.18, P=0.026), and in SLE patients with high PRL than in those with normal PRL (1.06±0.26 vs 0.82±0.21, P=0.005). However, there was no significant difference between SLE patients with normal PRL and healthy controls in regard to the expression of IRF-1 gene (P=0.514). The stimulation with rhRPL significantly elevated the relative expression of IRF-1 gene in SLE patients with normal PRL (0.99±0.22 vs 0.82±0.21, P=0.036), but had no obvious effect on that in the normal controls. Conclusion The study reveals a high expression of IRF-1 gene in SLE patients, which may be related to the high level of PRL.
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Objective To study changes of the autoimmune antibody level in patients after autologous hematopoietic stem cell transplantation(CD_(34)~+).Methods Twelve patients with SLE received autologous hematopoietic stem cell transplantation.All they survived and their immune system were recoverd after a period of time.The serum autoimmune antibody levels were measured before and after the transplantation,Results The antibody levels became normal 6 months after transplantation.Conclusion Autologous hematopoietic stem cell transplantation can effectively reduces the level of autoimmune antibody in patients with SLE.
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To investigate the predisposing role of HLA-DR genes to systemic lupus erythematosus (SLE), We used polymerase chain reaction and sequence specific oligonucleotide (PCR/SSO) probe hybridization to type HLA-DR subregion in the patients with SLE of Han nationality from Jiangsu province and matched control subjects. The results indicated that DR2 gene frequency was significantly more frequent in patients than that in controls. Whereas DR4 significantly decreased in patients compared with controls. It suggests that DR2 or the other unidentified genes tightly linked to it might be the susceptible genes of SLE. Whereas DR4 might have a protective effect on SLE.
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Objective Through studing the genotype and quantitatively analysing expression of type Ⅰ complement receptor(CR1) on erythrocytes from the patients with systemic lupus erythematosus(SLE). To investigate the relationship between CR1 and SLE. Methods Polymerase chain reaction(PCR) and Hind Ⅲ restriction enzyme digestion were used to analyse the genotype of CR1, and the quantitative expression of CR1 was demonstrated by FACscan flow cytometry. Results Compared with normal controls, high expression genotype of CR1 was significantly decreased, but low expression genotype was significantly increased in SLE patients. The number of CR1 on erythrocytes in SLE patients was significantly lower than that in normal controls. There was a negative relationship between the expression of CR1 and disease activity index of SLE (SLEDAI). Conclusion The expression of CR1 on erythrocytes might be related to the development of SLE, and it is helpful to detect the CR1 on erythrocytes for the determination of SLE patient′s condition.