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    基础研究

  • WANG Xin;YU Xue-qing;JIA Zhan-jun;CHEN Wen-fang;LI Xiao-yan;DOU Xian-rui;HAO Wen-ke;PENG Wen-xing;ZHANG Shi-hong;SUN Liao;HUANG Feng-xian;NIE Jing;LAN Hui-yao
    2007, 23(2): 76-81.
    Abstract ( 5246 ) PDF ( 889 ) Knowledge map Save
    Objective To explore the effect of overexpression of Smad7 on the peritonitis and peritoneal function. Methods Smad7 was upregulated in rat peritoneum by using ultrasound-microbubble system, and acute peritonitis was induced by intraperitoneal injection of E.Coli. Eighteen male SD rats were randomly divided into 3 groups: normal group, control group and model group. Three days after the transfection of empty plasmid or Smad7, acute peritonitis was induced by intraperitoneal injection of E.Coli ATCC 25922 109 CFU/kg. Rats were sacrificed at day 2(5 days after the transfection). WBC counting of the blood and peritoneal fluid, peritoneal pathological change and peritoneal function(PET) were measured. The expression of Smad7 was determined by Western blot and immunostaining respectively. The infiltration of inflammatory cells was determined by immunostaining of CD45. The bacterial clone forming unit(CFU) of peritoneal fluid was also detected. The relationships between peritoneal function and WBC or bacteria in peritoneal fluid were studied by SPSS 11.0. Results Smad7 overexpressed group had an increased WBC in the the peritoneal fluid and peritoneum with significant ultrafiltration failure and increased solute transport rate over peritoneum(glucose and total protein). Bacteria CFU in peritoneal fluid decreased significantly. Further analysis showed that peritoneal function was negatively correlated to the WBC counting of peritoneal fluid, without correlation to the amount of bacteria in the peritoneal cavity. Conclusion Overexpression of Smad7 in rat peritoneal membrane enhances peritoneal local inflammatory reaction, which helps to eliminate bacteria and hampers peritoneal function.
  • YE Kun;LIU Fu-you;LI Ying;PENG You-ming;DUAN Shao-bin;XU Xiang-qing;ZHOU Le-tian;LI Zhi-lan
    2007, 23(2): 82-86.
    Abstract ( 4480 ) PDF ( 843 ) Knowledge map Save

    Objective To investigate the effects of treatment with norcantharidin(NCTD) on proteinuria and renal pathology in protein-overload nephropathy rats. Methods Uninephrectomized SD rats received intraperitoneally injections of bovine serum albumin (BSA). The nephropathy rats were randomly divided into model group(BSA by intraperitoneally injection ) and NCTD group(BSA and NCTD 0.1 mg·kg-1·d-1 by intraperitoneally injection). An additional set of uninephrectomized rats only received saline as control group. Each group consisted of 8 rats. Twenty-four-hour urinary protein was measured weekly. At the end of the fifth week, the rats were sacrificed after anticoagulant blood and serum were collected and the remnant kidney of each rat was harvested and weighed for the examination of light microscopy, electron microscopy and immunofluorescence staining. Results Compared with control group, the ratio of kidney weight/body weight, urinary protein excretion, average score of glomerulosclerosis and tubulointerstitial lesion increased significantly(P < 0.01), but serum albumin(Alb) decreased obviously (P < 0.01)in model group. Electron microscope showed that mesangium region enlarged with obvious electron dense deposits, and foot process fusion was seen extensively in model group. Immunofluorescence of IgG and C3 with the intensity of 3+~4+ was also found in mesangium region of model group. After intervention with NCTD, the above-mentioned parameters, except that Alb enhanced significantly(P <0.01), all decreased significantly(P < 0.05 or 0.01). In NCTD group, few electron dense deposits and local foot process fusion were found, while immunofluorescence staining intensity of IgG and C3 weakened. There were no significant differences of serum creatinine(Scr), BUN and blood routine among all groups(P > 0.05). Conclusion NCTD may decrease the proteinuria of protein-overload nephropathy rats and attenuate tubulointerstitial lesion without severe side effects of liver, kidney or bone marrow.

  • JIN Ying-shun;JIN Xiu-nan;JIN Hua;CUI Zhen-hua;CHEN Ying;LI Can
    2007, 23(2): 87-90.
    Abstract ( 4169 ) PDF ( 761 ) Knowledge map Save
    Objective To investigate the renoprotective mechanism of losartan in a rat model of chronic cyclosporine A(CsA) nephrotoxicity. Methods Adult Sprague-Dawley rats kept on a salt-depleted diet (0.05% sodium) were treated daily for four weeks with vehicle (olive oil, 1 ml/kg), CsA (15 mg/kg) or both CsA and losartan (10 mg/kg in drinking water). The effect of losartan on NF-κB and βig-h3 was evaluated using electrophoretic mobility shift assay (EMSA) and immunoblotting. Histopathology and renal function were compared among three groups. Results Compared with the vehicle-treated rats, rats given CsA showed an increase of NF-κB activity [(201±15)% vs(104±7)%, P < 0.01], a decline of IκB expression, and an increase of βig-h3 protein expression [(300±35)% vs(105±15)%,P < 0.01). Concurrent administration of losartan reversed all of above parameters (P < 0.05). There were no significant differences in renal function among the tree groups. Conclusion Losartan is capable of abrogating the activation of NF-κB and βig-h3, and this is associated with attenuating tubulointerstitial fibrosis in chronic CsA nephrotoxicity.
  • LI Yan-bo;HAN Jun-yong;LI Wei-min
    2007, 23(2): 91-94.
    Abstract ( 3555 ) PDF ( 1161 ) Knowledge map Save
    Objective To investigate the relationship between p38 mitogen-activated protein kinase (p38MAPK) and transforming growth factor-β (TGF-β), and to explore the effect mechanism of p38MAPK on diabetic nephropathy (DN). Methods Rat mesangial cells (RMC) were separately incubated with high glucose, advanced glycosylation end products (AGE) and H2O2 after pre-reatment of SB203580 (p38MAPK special inhibitor) and simvastatin. The protein expression of p-p38MAPK and TGF-β was examined by Western blot. Results Compared to the control group, high glucose, AGE and H2O2 significantly activated p38MAPK and increased the protein expression of p-p38MAPK and TGF-β (P < 0.05). The protein expression of TGF-β was significantly inhibited by SB203580 (P < 0.05). Compared to RMC incubated with corresponding stimulations, p-p38MAPK and TGF-β were expressed at a lower level in RMC pre-treated with simvastatin (P < 0.05). Conclusions p38MAPK may be one of the initial onset signals of diabetic nephropathy. Simvastatin may effectively attenuate DN progression through down-regulating the expression of TGF-β via p38MAPK.
  • ZHANG Xin-ping;YI Zhu-wen;HE Xiao-jie;HE Qing-nan;ZHU Min;ZHOU Gang
    2007, 23(2): 95-100.
    Abstract ( 3262 ) PDF ( 887 ) Knowledge map Save
    Objective To examine the influence of antisense MCP-1 on MCP-1 production and mesangial cell proliferation. Methods The rat glomerular mesangial cells were infected with retroviral supernatants of pLXSN-A-MCP-1 containing antisense MCP-1 and verified by Sorthern blot analysis and PCR. Lipopolysaccharide(LPS) stimulated the infected mesangial cells. Cell growth was measured by cell counting method. MCP-1 expression was detected by RT-PCR and enzyme-linked immunosorbent assay(ELISA). Results Southern blot analysis and PCR demonstrated that antisense MCP-1 cDNA-infected mesangial cells were established successfully. Treated with LPS, the growth rate of infected cells was significantly lower than that of normal cells. The level of MCP-1, CCR2 mRNA formation and the level of MCP-1 protein secretion decreased compared with those of normal cells. Conclusions pLXSN can transfer MCP-1 cDNA into mesangial cells. Antisense MCP-1 decreases MCP-1 transcription and translation. Antisense MCP-1 decreases the proliferation of mesangial cells in inflammation. CCR2 also expresses in mesangial cells. MCP-1 is a autocrine chemokine.
  • LI Ying;DUAN Hui-jun;ZHANG Tao;LIU Mao-dong;SHI Yong-hong;LIN Hai-ying;WANG Bao-xing
    2007, 23(2): 101-105.
    Abstract ( 3170 ) PDF ( 951 ) Knowledge map Save
    Objective To observe the activity of extracellular signal-regulated kinase(ERK) in glomerular mesangial cells(GMC)exposed to high glucose and the effect of valsartan on ERK activity, and explore the possible renoprotection mechanism of valsartan.Methods GMC were cultured in vitro and the groups were designed as follows: low glucose group(NG, 5.5 mmol/L d-glucose), high glucose group(HG, 30 mmol/L d-glucose), valsartan group(HG+val, 30 mmol/L d-glucose+valsartan 10 μmol/L) and mannitol group(MG, 24.5 mmol/L mannitol +5.5 mmol/L d-glucose). The expression of phosphate (p-) ERK1/2 protein was detected by immunocytochemical technique and Western blot. RT-PCR and radioimmunoassay were respectively used to measure the expression of TGF-β1 mRNA in mesangial cells and the content of type Ⅳ collagen in supernatant. Results The strongly positive staining of p-ERK1/2 in mesangial cells exposed to high glucose was observed not only in cytoplasm, but also in nuclei at 12 h, with an increasing tendency to 72 h (P < 0.01). The level of TGF-β1 mRNA in mesangial cells and the content of type Ⅳ collagen in the supernatant of HG group were also significantly higher than those of NG group (P < 0.01). Compared with those in HG group, the above parameters were significantly down-regulated in HG+val group (P < 0.01), whereas there were no significant differences of above parameters between MG group and NG group. Conclusion The expression of p-ERK1/2 is up-regulated in GMC exposed to high glucose medium, which can be inhibited by valsartan.
  • 透析与移植

  • TANG Li-jun;CHEN Hui-min;CHENG Li-tao;GU Yue;WANG Tao
    2007, 23(2): 106-109.
    Abstract ( 4556 ) PDF ( 1137 ) Knowledge map Save
    Objective To investigate the association between E/T (extracellular water to total body water ratio) and pulse wave velocity (PWV) in patients on continuous ambulatory peritoneal dialysis(CAPD). Methods Clinical stable CAPD patients (n=56,26M/30F) in one single center were included. Carotid-femoral PWV was measured with a validated automatic device and was used as an index of large arterial stiffness. Multiple-frequency bioelectrical impedance analysis was used to record the values for ECW, intracellular water (ICW), and total body water. Based on these data, E/T was also calculated. In addition, some biochemical indices, such as serum albumin, blood urea nitrogen, serum creatinine, serum triglycerides, total cholesterol lipoprotein, low-density lipoprotein, high-density lipoprotein, alanine and aspartate transaminase, etc were determined with standard methods. Pearson’s correlation and multiple regression analysis were performed to identify the relationship between E/T and PWV. Results PWV was strongly associated with E/T (r = 0.454, P = 0.001), E/I(r = 0.456, P = 0.001), pulse pressure(PP)(r =0.649, P < 0.01),age(r = 0.404, P = 0.002), serum albumin(r= -0.346, P = 0.01)and CRP(r =0.327, P = 0.025), respectively. Multiple regression analysis showed that PWV was independently determined by E/T(β= 0.472, P = 0.001), PP(β= 0.442, P = 0.001) and CRP(β= 0.246,P = 0.05). They accounted for 58.1% of the total variance and E/T alone represented 37.8% of the explained variance. Conclusions E/T is closely associated with PWV in CAPD patients. E/T, in addition to PP and CRP, is an independent risk factor for elevating PWV in CAPD patients, suggesting that the increase of arterial stiffness may be the link between fluid overload and cardiovascular events and mortality in CAPD patients.
  • RONG Shu;YE Chao-yang;SUN Li-jun;CHEN Jing;ZHANG Bin;MEI Chang-lin
    2007, 23(2): 110-112.
    Abstract ( 6200 ) PDF ( 1141 ) Knowledge map Save
    Objective To observe the efficacy and safety of 46.7% sodium citrate versus heparin catheter lock for permanent hemodialysis catheters.Methods Forty-one maintenance hemodialysis patients with permanent catheters were divided into 2 groups randomly. The patients in citrate group received 46.7% sodium citrate lock for catheters after each dialysis session for 6 months. The patiency, catheter-related infection, bacterial culture of catheter lock solution and predialysis serum electrolytes were compared between two groups. Results The impatiency rate and catheter-related infection rate of citrate group were significantly lower than those of heparin group and pretreatment(P < 0.05). The serum electrolytes did not change markedly. In citrate group, the positive rate of catheter lock solution bacterial culture decreased after 2 months, and became negative after 4 months. The main side effect in citrate group was light oral lips numbness (1.93%). Conclusions Catheter filling with a solution containing 46.7% sodium citrate may significantly reduce the incidence of impatency and catheter-related infection. It appears to be effective and safe and does not carry the risk of severe side effects of high concentrations of citrate.
  • 新技术与方法

  • JIN Xiao-ming;ZHANG Lei;SHI Jiong;HUANG Li-juan;TONG Dan-dan
    2007, 23(2): 115-120.
    Abstract ( 6114 ) PDF ( 1386 ) Knowledge map Save
    Objective To establish an optimal experimental animal model of IgA nephropathy (IgAN). Methods Three kinds of proteins, Dextran G200, outer membrane proteins (OMPs) of Bacillus coli (E.coli) and 20 peptide cellular antigen determinant of staphylococcus aureus, were used to induce IgAN animal model respectively. Animal models were identified and their clinicopathological features were compared by molecular biological and pathological methods. Results (1) In dextran G200 group, urinary protein elevated obviously accompanied by hematuria; immunofluoresence staining revealed a great quantity of IgA depositing in some glomeruli; light microscopy showed a remarkable proliferation of mesangial cells in kidney and diffuse pink-dyed substance deposition in liver and spleen; electron microscopy demonstrated a low electron dense deposition in glomerular mesangial region and amylonloid material in liver and spleen. (2) In OMPs group, urinary protein elevated obviously accompanied by hematuria; immunofluoresence staining revealed less amount of IgA deposition in glomeruli; light microscopy showed a slight proliferation of mesangial cells and obvious interstitial infiltration of inflammatory cells; no electron dense deposition was found in glomerular mesangial region by electron microscope. (3) In 20 peptide cellular antigen determinant of staphylococcus aureus group, urinary protein elevated obviously accompanied by hematuria as well; immunofluoresence staining revealed a large quantity of IgA deposition in most glomerui; light microscopy showed a slight proliferation of mesangial cells and matrix; electron microscopy demonstrated a high electron dense deposition in subendothelial cells and mesangial region. Conclusion The clinicopathological characteristics of IgAN animal model induced by 20 peptide antigen determinant of staphylococcus aureus are similar to human IgAN, which is the best among these three animal models.