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Objective To observe the effect of recombinant adenovirus-mediated the fms-like tyrosine kinast-1 (FLT-1) gene promoter carrying protein C (PC) gene (Ad-FLT-1/PC) on apoptosis and oxidative stress in rat with diabetic nephropathy atherosclerosis. Methods High sugar, high fat diet and streptozotocin by intraperitoneal injection were used to establish diabetic nephropathy atherosclerosis rat model. The rats were randomly divided into Ad-FLT-1/PC group (n=23), Ad-GFP group (n=23) and normal saline group (n=22) and injected with 300 ul Ad-FLT-1/PC, Ad-GFP and normal saline by caudal vein. Another healthy rats (n=23) were control group. At day 1, 3, 7 and 14 after transfection, rats of each group were randomly executed to observe vascular apoptosis and assay the level of SOD and MDA in plasma by the kit method of WST and TBA. Results Vascular cell apoptosis was observed in Ad-FLT-1/PC group, Ad-GFP group and NS group, the apoptosis index showed no statistical difference between three groups at each time point (P>0.05). At day 14 after transfection, Ad-FLT-1/PC group rats had higher concentration of the plasma SOD and lower MDA than Ad - GFP group and NS group. The difference were statistically significant (all P<0.05). Conclusions The recombinant adenovirus Ad-FLT-1/PC can effectively regulate oxidative stress but not apoptosis.
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Objective To explore the role of liver X receptor (LXR) agonist T0901317 on thrombomodulin (TM) expression in human glomerular endothelial cells and the possible mechanisms.Methods Different concentrations of T0901317 were used to stimulate human glomerular endothelial cells for different time,then LXRα,LXRβ expression were detected by using Western blotting analysis;the roles of T0901317 on TM mRNA and TM protein expression were observed by using real-time PCR,Western blotting and immunofluorescence assay.LXRα,LXRβ gene interference segment Si-hLXRα,Si -hLXRβ were transfected into human glomerular endothelial cells with the concentration of 100 nmol/L respectively,then the roles of Si-hLXRα,Si-hLXRβ on the TM protein and TM mRNA expression were assayed by Western blotting and real time PCR.Results Human glomerular endothelial cells expressed LXRα and LXRβ.Compared to the normal cells and DMSO group,T0901317 could significantly promote TM expression in human glomerular endothelial cells (P ＜ 0.05) and showed a time -and dose-dependent manner.TM expression in Si-hLXRα transfected group was significantly lower than that in the control group (P ＜ 0.05),while TM expression in Si-hLXRβ transfected group had no significant difference compared to the control group.Conclusions Human glomerular endothelial cells express LXRα and LXRβ.LXR agonist T0901317 promotes TM expression in human glomerular endothelial cells,which may be mainly through activating LXRa.
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ObjectiveTo investigate the effects and molecular mechanism of endoplasmic reticalam stress (ERS) on albumin-induced apoptosis in renal proximal tubular cells (HKCs). MethodsWestern blot was performed to detect the relationship of the expression of glucose-regulated protein 78 (GRF78) and CCAAT/enhancer-binding protein-homologous protein (CHOP) with the action time and concentration of haman serum albumin (HSA). Expression levels of CHPO mRNA and protein in HKCs after CHOP siRNA transfection were examined by real-time fluorescence quantitative PCR and Western blot respectively. Annexin-V-FITC and PI doable staining cytometry was used to detect the apoptosis of HKCs induced by HSA and influenced by CHOP siRNA. Results(1)After HKCs were stimulatde by 0, 5, 10, 20 g/L albumin for 24 hours respectively, the expression of GRP78, CHOP and HKCs apoptosis were increased with the albumin concentration (P<0.01). After HKCs were stimulated by 20 g/L albumin for 0, 6, 12, 24, 36 hours respectively, the expression of GRP78 was up-regulated at 6-hour, while CHOP and HKCs apoptosis were increased at 12-hour, and significant differences were found among groups (P<0.01). (2) CHOP siRNA significantly inhibited albumin-induced HKC CHOP mRNA and protein expression, as well as HKC apoptosis (P<0.01). ConclusionsRenal tubular cells exposed to high protein load result in EBS. ERS may subsequently lead to tubular damage by activation of pro-apoptosis factor CHOP.
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Objective To explore the influence of albumin-activated renal tubular epithelial cells (RTECs)on peritubular capillaries in co-culture system and its potential mechanism. Methods Endocytosis of TRITC labeled bovine scrum albumin (TRITC-BSA) by HKC was detected by laser scanning confocal fluorescence microscope. HKC or HKC transfected with cubilin (endocytic receptor of albumin) siRNA or pre-treated with rotenone was incubated with albumin(20 g/L) for 24 h respectively. Fluorescence probe technique and spectrometry were applied for determination of intracellular superoxide anion O2-and H2O2 in supematant. Then, the albumin-aetivated-HKC, pretreated-HKC with cubilin siRNA or rotenone, was cultured with HUVEC for 24 h in co-culture system respectively. HUVEC proliferation was determined by MTT and cellular apoptosis was analyzed by flow cytometry. Tabular morphogenesis of endothelial cells was examinedby microscopy. Results TRITC-BSA uptake was obviously lower in HKC transfected with cubilin siRNA. Intracellular generation of O2-and H2O2 in culture supernatant was increased in dose-and time-dependent manner after stimulating with albumin. The levels of O2-and H2O2 were suppressed by cubilin siRNA and rotenone. In co-culture system, albumin-activated-HKC induced endothelial cells apoptosis and inhibited their capillary tubular morphogenesis. Pretreatment of HKC with cubilin siRNA or rotenone could suppress endothelial cells apoptosis and promote capillary tubular morphogenesis. Conclusions There may be a crosstalk between RTECs and peritubular microvascular endothelial cells in renal proteinurie diseases. The generation of ROS by albumin-activated RTECs may play an important role in this process.
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Objective To explore the role of delivering programmed death (PD)-1 negative costimulation pathways in the prevention of murine BXSB lupus. Methods A recombinant adenovims containing the full-length mouse PD-L1 gene (AdPD-L1) was constructed and was introduced to the BXSB mice. The effect of immunoinhibitory receptor PD-1 on activated lymphocytes was investigated to evaluate its effect on prevention of lupus nephritis in BXSB mice. Results This intervention dramatically delayed the onset of proteinuria, effectively inhibited IgG autoantibody production, and significantly reduced hypercellularity and deposition of IgG in glomeruli, resulting in almost complete amelioration of lupus nephritis in these animals.Conclusion Our results suggest that simultaneous stimulation of PD-1-mediated pathway can potentially revent human lupus nephritis.
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OBJECTIVE: To investigate the curative effects and side effects of hirudin in treating immunoglobulin A nephropathy (IgAN) with hematuria and minimal proteinuria in a short-term. METHODS: Two hundred and sixty-two histologically confirmed cases of IgAN with hematuria and minimal proteinuria from 1998 to 2007 were randomly divided into hirudin-treated group (peroral administration of Maixuekang capsules) and dipyridamole-treated group (peroral administration of dipyridamole). In the two groups, contrast analysis of conformation and counts of erythrocytes in urine, urine protein quantitation in 24 hours, levels of serum creatinine (Scr) and creatinine clearance rate (Ccr), blood lipid, five items of blood clotting and side effects was performed. RESULTS: After six-month treatment, the anisotrophy rate and the counts of erythrocytes in urine, and the urine protein quantitation in 24 hours in hirudin-treated group were decreased distinctly as compared with pre-treatment (P<0.01) and dipyridamole-treated group (P<0.05). On the other hand, Ccr was increased obviously in hirudin-treated group as compared with pre-treatment and dipyridamole-treated group (P<0.01). The blood lipid was also ameliorated in hirudin-treated group, but there was no significant difference. The anticoagulation effect of hirudin was better than dipyridamole (P<0.01). Efficacy assessment showed that the total response rate, complete remission rate and predominance remission rate in hirudin-treated group were higher than those in dipyridamole-treated group. Few side effects were found in both groups, and the rate of adverse reaction in gastrointestinal tract was lower in hirudin-treated group as compared with that in dipyridamole-treated group (P<0.05). CONCLUSION: Compared with dipyridamole, hirudin has superiority in kidney protection and decreasing the anisotrophy rate, counts of erythrocytes in urine and the urine protein.
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Objective To investigate the expression of programmed death-ligand 1 (PD-L1) and programmed death-1(PD-1) in the kidney tissue of acute allograft rejection (AR) and the correlation between PD-L1 expression and PD-1 expression and the degree of tubulointerstitial lesions.Methods Immunohistochemistry and in situ hybridization were used to detect the expression of PD-L1 and PD-1 in renal tissue of normal control and AR confirmed by biopsy.Results Both PD-L1 and PD-1 expression were significantly higher in AR than in the normal renal tissue (P
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Objective To investigate the role of PD-L1 in the pathogenesis of acute allograft rejection (AR) and its correlation with PD-1 and the degree of tubulointerstitial lesions. Methods A total of 20 patients admitted by our hospital and Daping hospital from 1999 to 2003 were included, 12 male and 8 female, aged 18-40 years old (average 32.2?4.5). All of them were diagnosed as acute rejection by renal biopsy. Fifteen specimens from the mismatched donor kidney and the normal tissues from the resected kidney served as normal control. Immunohistochemistry was used to detect the expression of PD-L1 and PD-1 in renal tissues of AR and normal control. Results Both PD-L1 and PD-1 expression were significantly higher in AR than that in normal control. The intensity of PD-L1 positive in renal tissues of AR showed a positive correlation with PD-1 expression and a negative correlation with the degree of tubulointerstitial lesions. Conclusion Increased expression of PD-L1 and PD-1 in renal tissues of AR has an important effect on the degree of tubulointerstitial lesions.