ABSTRACT
Objective:To investigate the expressions of transforming growth factor-β1 (TGF-β1), aquaporin (AQP) and osteopontin (OPN) in renal injury induced by fluorosis in rats.Methods:According to body weight (80 - 100 g), forty-eight SD rats were divided into control group (normal saline), low fluoride group (10 mg/kg) and high fluoride group (20 mg/kg) by random number table, 16 rats in each group (half males and half females). The rats were exposed to fluoride by intraperitoneal injection of sodium fluoride. After the rats were treated with fluoride for 12 weeks, the rats were killed by femoral artery bloodletting and the renal tissue was taken. The contents of TGF-β1, AQP and OPN in serum of rats with fluorosis were analyzed by enzyme-linked immunosorbent assay (ELISA). The expressions of TGF-β1, AQP and OPN in renal tissue of rats with fluorosis were analyzed by quantitative real-time PCR (qRT-PCR) and immunocytochemistry.Results:The serum levels of TGF-β1, AQP and OPN in high fluoride group [(26.42 ± 6.09), (378.60 ± 36.84) μg/L, (603.45 ± 64.32) pg/ml] were higher than those in control group [(2.41 ± 0.42), (157.41 ± 15.26) μg/L, (182.45 ± 30.63) pg/ml] and low fluoride group [(13.15 ± 3.26), (245.65 ± 23.21) μg/L, (359.47 ± 55.26) pg/ml, P < 0.05]. The levels of TGF-β1, AQP and OPN in low fluoride group were higher than those in control group ( P < 0.05). The expressions of TGF-β1, AQP and OPN mRNA in high fluoride group were higher than those in control group and low fluoride group ( P < 0.05), and the expressions of TGF-β1, AQP and OPN mRNA in low fluoride group were higher than those in control group ( P < 0.05). The positive expression scores of TGF-β1, AQP and OPN in high fluoride group were higher than those in control group and low fluoride group ( P < 0.05), and the positive expression scores of TGF-β1, AQP and OPN in low fluoride group were higher than those in control group ( P < 0.05). Conclusion:TGF-β1, AQP and OPN are highly expressed in the renal tissue of rats with fluorosis, which can be used as indicators to judge the damage of renal aggregation system caused by fluorosis.
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Objective To investigate the effects of ch1D1, an anti-CD86 chimeric antibody, on autoreactive B lymphocytes isolated from patients with systemic lupus erythematosus ( SLE) . Methods Flow cytometry analysis was performed to measure the expression of CD86 on the surface of B cells isolated from patients with SLE and to analyze the effects of ch1D1 on the activation of CD4+T cells. The method of magnetic bead sorting was used to separate B cells, NK cells and CD4+T cells from PBMC collected from healthy subjects and patients with SLE for subsequent experiments. Antibody-dependent cell-mediated cyto-toxicity (ADCC) and complement-dependent cytotoxicity (CDC) that were mediated by ch1D1 were meas-ured with LDH release assay. Effects of ch1D1 on the secretion of auto-antibodies and the proliferation of CD 4+ T were detected by ELISA and 3 H -thymidine ( 3 H-TdR) incorporation assay, respectively. Results The levels of CD80 (68. 08±14. 28 vs 46. 10±12. 14, n=24, P<0. 000 1) and CD86 (44. 72±14. 90 vs 13. 99±10. 74, n=24, P<0. 000 1) expressed on the surface of B cells isolated from patients with SLE were significantly higher than those from the healthy subjects, suggesting the abnormal activation of B cells. Com-pared with the negative control group and the murine monoclonal antibody 1D1, ch1D1 was more effective in mediating the ADCC and CDC responses (P=0. 017 2, P=0. 038 8). Activated T cells significantly en-hanced the secretion of auto-antibodies by B cells isolated from patients with SLE. Compared with the nega-tive control group, the enhanced secretion of auto-antibodies was significantly inhibited by treatment with ch1D1 (P=0. 001 9). Moreover, ch1D1 significantly inhibited the proliferation and activation of CD4+T cells induced in patients with SLE (P=0. 002 4, P=0. 049 5). Conclusion ch1D1, the anti-CD86 chim-eric antibody, could effectively mediate the ADCC and CDC responses against autoreactive B cells isolated from patients with SLE, inhibit the secretion of auto-antibodies and suppress the proliferation and activation of auto-reactive CD4+T cells. It might be a potential immunotherapy agent for the treatment of SLE.
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Objective To explore the effects of zinc finger protein A20(A20) on the expression of CD40 and the phosphorylation of IκBαas well as the local inflammatory reaction of abdominal cavity in Sprague Dawley (SD) rats with peritoneal dialysis-related a-cute peritonitis induced by lipopolysaccharide (LPS ) .Methods 24 male SD rats were equally randomized to four groups (n= 6 , each) .Control group:injected with 4 .25% dextrose peritoneal dialysate(PDF) via abdominal cavity (90 mL/kg);LPS group :injec-ted with LPS(1 mg/kg) via abdominal cavity 4 hour later followed by PDF injection ;transfection A20 plasmid group and empty plasmid group:after transfer pGEM-T easy-A20 or pGEM-T easy plasmid via intraperitoneally using an ultrasound-microbubble-mediated system for 3 days ,then injected with LPS and PDF via abdominal cavity .The rats were killed 4 hours after PDF injection . Peritoneum tissue was stained using Masson and HE .Leucocytes count in abdominal dropsy was performed .The proteins expres-sion of A20 ,p-IκBα,IκBα,and CD40 in peritoneum tissue were analyzed by western blot ;the expression of CD40 mRNA in peritone-um tissue were determined by RT-PCR;IL-6 level in abdominal dropsy was determined by ELISA .Results LPS could induce the protein expression of A20 in rats peritoneum tissue .Compared with LPS group and empty plasmid group ,the degree of edema ,in-flammatory cells infiltration ,and the ratio of p-IκBα/IκBα,mRNA and protein expression of CD40 in rats peritoneum as well as leu-cocyte counts and IL-6 level of abdominal dropsy were also significantly decreased in transfection A20 plasmid group(P0 .05) ,but the mRNA and protein expression of CD40 were significantly higher than that of control group (P<0 .05) .Conclusion Over expression of A20 in SD rats peritoneum tissue could down-regulate the inflammatory reaction in peritonitis induced by LPS ,which might be involved in modulating the expression of associated functional protein during LPS signal pathw ay .
ABSTRACT
Objective: To study the relation of plasma Se to oxidative stress in the patients with primary glomerular diseases (PGS) and its clinical significance. Methods: Plasma Se concentration ,GSH-Px and SOD activities and contents of MDA in 45 patients with PGS and 20 normal control (NC)were measured by catalytic polarography and colorimetric assay. Serum creatinine, urea nitrogen and uric acid concentrations in these patients were determined simultaneously. Results: 1. Plasma Se contents of both PGS patients of normal(groupⅠ) and impaired renal function (groupⅡ) were significantly lower than those in NC;2. PGS patients of groupⅠandⅡhad significantly lower plasma GSH-Px and SOD activities than NC and their MDA contents were siginificantly higher. 3. Plasma GSH-Px and SOD activities and MDA contents between groupⅠand Ⅱ were not different; 4. Multiple logistic regression analysis showed that plasma Se content in patients with PGS was positively related to plasma GSH-Px , SOD activities and negatively correlated with MDA. Serum creatinine concentrations were negatively correlated significantly with plasma Se, GSH-Px, SOD and positively related to MDA. Conclusion: Plasma Se deficiency may be an important non-immune factor resulting in or worsening PGS through weakening body抯 antioxidative defence. It is suggested that sufficient Se supplementation in practice may be obviously beneficial to oxidative stress in the patients with PGS.
