ABSTRACT
Objective To investigate the regulation of mesenchymal stem cells (MSCs) on the expression of tumor necrosis factor-α-induced protein 8-like 2 (TNFAIP8L2, TIPE2) in the peripheral blood-derived macrophages of systemic lupus erythematosus (SLE) patients. Methods The expression of TIPE2 mRNA in peripheral blood mononuclear cells (PBMCs) and macrophages from SLE patients were detected by Quantitative polymerase chain reaction (qPCR). The TIPE2 protein levels in SLE PBMCs and peripheral blood-derived macrophages were detected by western blot, flow cytometry and immunofluorescence staining, respec-tively. Data were analyzed by t test and Spearman correlation analysis. Results The TIPE2 mRNA expres-sion in PBMCs of SLE patients was significantly lower than that of healthy controls [(0.41 ±0.14) vs (1.06±0.39), t=5.376, P<0.01], as well as the TIPE2 protein level [(0.40 ±0.21) vs (1.09 ±0.26), t=2.963, P<0.05]. The expression of TIPE2 mRNA in peripheral blood-derived macrophages from SLE patients was significantly decreased [(0.56±0.24) vs (1.07±0.38), t=5.203, P<0.01). Moreover, TIPE2 mRNA level of peripheral blood-derived macrophages was negatively correlated with systemic lupus erythematosus disease activity index (SLEDAI) score (r=-0.60, P<0.01), 24-hour urinary protein (r=-0.46, P<0.05) and erythrocyte sedimentation rate (r=-0.46, P<0.05) in SLE patients. The percentage of TIPE2+cells in peripheral blood-derived macrophages (TIPE2+/CD14+)％ from SLE patients was significantly lower than that in healthy controls [(51.4 ±18.5)％ vs (82.4 ±7.5)％, t=2.679, P<0.05]. After 24 hours co-cultured with MSCs, the TIPE2 mRNA expression in SLE per-ipheral blood-derived macrophages was significantly increased [(2.2 ±0.7) vs (1.0 ±0.3), t=3.729, P<0.05). Immunofluorescence results showed the same increase of TIPE2 protein in SLE peripheral blood-derived macrophages [(0.112 ±0.020) vs (0.074 ±0.016), t=3.268, P<0.05]. Conclusion The TIPE2 level in peripheral blood-derived macrophages of SLE patients are decreased. MSCs upregulate the TIPE2 expression in vitro, suggesting that TIPE2 can be a new target for MSCs in the treatment of SLE.
ABSTRACT
Objective To investigate the effects of diammonium glycyrrhizinate (DG) on Toll-like receptor 4 in rat alveolar epithelial cells induced by paraquat (PQ).Methods Rats in the PR and PDR groups were induced 2 h by RU486 (100 nmol/L).Then rats in the DG and PDR groups were induced 2 h by DG (0.6 mg/mL).Finally,PQ (0.6 mg/mL) were administered and induced 24h in the PQ,DG,PR and PDR groups,while,the NE group was induced 24 h by absolute ethyl alcohol (0.33 μ mol/L),and the NS group was induced 24 h without drugs.MTT assay was used to measure the cell growth and inhibitioneffects of PQ and DG on cells.The ELISA assay was applied to measure the levels of the TLR-4,Myd88,NF-κB P65 and GR.The gene expressions ofTLR-4,Myd88,NF-κB P65 and GR were detected by RT-PCR.Results The survival rate of rat alveolar epithelial cells was decreased by PQ (200,400,600,800,1 000,1 500,2 000 μmol/L),and the IC50 value for 24 h was 927.045 μmol/L.The inhibition rates were (11.74±1.44)％,(18.76±1.30)％,(28.74±0.54)％,(40.30±0.55)％,(51.24±0.76)％,(68.19±1.10)％,(83.16±0.59)％ in the 200,400,600,800,1 000,1 500,2 000 pmol/L PQ treatment groups,respectively.And the inhibition rates were (48.01±1.37)％,(40.68±2.33)％,(32.76±4.11)％,(34.12±4.3)％,(39.22±2.23)％,(51.26±-0.39)％ in the 0.2,0.4,0.6,0.8,1,and 2 mg/mL DG treatment groups,respectively.The levels of TLR4,Myd88,NF-κB P65,and TNF-a in the PQ and PR groups were higher than those in the NS group (all P＜0.01).While,the levels of GR in the PQ and PR groups were lower than that in the NS group (all P＜0.01).And,the levels of TLR4,Myd88,NF-rκB P65,and TNF-α in the DG and PDR groups were lower than those in the PQ group (all P＜0.01).But the levels of GR in the DG and PDR groups were higher than that in the PQ group (all P＜0.01).Conclusions Diammonium Glycyrrhizinate can attenuate the injury of rat alveolar epithelial cells induced by paraquat,can decrease the levels of TLR-4,Myd88,NF-KB and TNF-α,and increase the GR level.