【Abstract】 Objective To establish a prediction formula for screening atherosclerotic renal artery stenosis (ARAS). Methods Clinical data of 892 patients who suffered from coronary artery disease (CAD) with renal artery angiography were reviewed. The clinical characters related to ARAS were analyzed and Logisitc regression was used to reveal the ratio of every clinical character. Based on the ratio, a prediction formula was established. Then sensitivity and specificity of this prediction formula was examined back to the patients. Results The patients who suffered from ARAS in CAD population accounted for 12.7%. The risk factors included age, BMI, Scr, hypertension history, diabetes mellitus (DM)，stroke and refractory hypertension. Age, BMI and Scr were used as continuous variables and other four as dichotomized variables. According to the prediction formula, the score was from 5.5 to 20.5. With the score increasing, the probability of ARAS increased as well. Conclusion This prediction formula is helpful to select the ARAS patients from CAD population, in order to provide them the evidence for further examination.

Abstract】 Objective To investigate the effects of geranylgeranylacetone (GGA) on tubulointerstitiai fibrosis in unilateral ureteral obstruction (UUO) rats. Methods Fifteen male Sprague-Dawley rats weighing 200～250 g were randomly divided into three groups: sham group, model group and GGA group. Rats subjected to UUO operation were treated with daily oral dose of vehicle(5% gum Arabic and 0.008% tocopherol) or 400 mg/kg GGA one day before UUO operation until being sacrificed. Five rats were sacrificed at day 7 in each group. Routine kidney pathology was examined. Expression of E-cadherin and α-SMA protein was assessed by indirect immunofluorescence and Western-blot. Tubular apoptosis was detected by determinal deoxynucleotidyl transferase (TdT)-mediated deoxyurideine triphosphated (dUTP) nick-end labeling (TUNEL) assay. Proliferating cell nuclear antigen (PCNA) was examined by immunohistochemistry. Results As compared to model group, treatment with GGA markedly elevated HSP72 level in kidneys, significantly reduced tubular lesions and attenuated interstitial fibrosis(48.7%±1.3% vs. 65.8%±7.3%, score 0.40±0.08 vs. 1.36±0.50, P<0.05); EMT was reduced by up-regulation of E-cadherin, down-regulation of α-SMA (P<0.05), and apoptotic tubular cells as well as proliferative tubular cells were decreased(6.78±1.25 vs. 2.81±0.63，57.61±5.42 vs. 17.66±1.38， respectively, all P<0.05) in GGA group. Conclusion GGA can attenuate tubulointerstitial fibrosis in rat UUO model, at least in part, by inhibiting tubular apoptosis.

【Abstract】 Objective To construct eukaryotic expression vectors carrying the small hairpin RNA (shRNA) for NG2 mRNA and to study the effect on proliferation of rat mesangial cells (RMC) induced by high glucose stimulation through transfection and expression of shRNA targeting NG2 gene. Methods RMC was cultured in vitro and treated with high glucose (30 mmol/L) for 24 h. The change of NG2 expression was observed. Oligonucleotide hairpin sequences targeting NG2 gene were designed by the internet tool and was then inserted into the plasmid Pgenesil-1 containing enhanced green fluorescence protein (EGFP) to form plasmid Pgenesil-siNG21 and Pgenesil-siNG22. Plasmid expressing irrelevant shRNA was used as negative control named Pgenesil-siNC. RMC was transfected with above three kinds of plasmids respectively. Then the cells were stimulated with high glucose, and the changes of cell proliferation were observed by MTT and flow cytometry (FCM) methods. Results The expression of NG2 mRNA in RMC was significantly increased (P<0.01) after 24 h treatment with high glucose. The expression of EGFP gene was (50±10)%. The expression of NG2 in RMC was significantly inhibited by Pgenesil-siNG21 and Pgenesil-siNG22. Compared with the control group, cell proliferation levels were markedly decreased in siNG2 following high glucose stimulation (P<0.01). Conclusion shRNA targeting NG2 gene can inhibit the cell proliferation of RMC induced by high glucose stimulation.

【Abstract】 Objective To investigate the effect of connective tissue growth factor (CTGF) on integrin-linked kinase (ILK) expression and its relationship with activation of MAPK, PI3-K signaling pathways in renal proximal tubular epithelial cells (PTEC). Methods Time and dose response of CTGF-stimulated ILK protein expression was measured in PTEC. After 24 h of serum depletion, the cells were incubated in serum-free medium containing different concentrations of CTGF (0, 1, 5, 20, 50 μg/L) for 24 h or were incubated in serum-free medium containing 50 μg/L CTGF for different times (0, 6, 24, 48, 72 h). The cells were pretreated with MEK inhibitor PD98059 (10 μmol/L), PI3-K inhibitor LY294002 (5 μmol/L) and P38 MAPK inhibitor SB203580 (10 μmol/L) respectively for 45 min before adding CTGF, then the cells were incubated for additional 12 h (real-time PCR) or 24 h (Western blotting). Results CTGF up-regulated ILK expression in a concentration-dependent manner, with a plateau at 50 μg/L by 5.638-fold as compared to the control (P<0.05). CTGF stimulated expression of ILK in a time-dependent manner as well. After exposure to CTGF (50 μg/L), the maximal level of ILK expression was reached at 48 h by 5.740-fold as compared to the control (P<0.05). Blockade of MAPK pathway and PI3-K pathway with PD98059 and LY294002 respectively could markedly inhibit CTGF-induced ILK expression (ILK mRNA CTGF：5.130±0.920， CTG＋PD98059：3.006±0.477， CTGF＋LY294002：3.700±0.387， all P<0.05). Inhibition of p38 MAPK pathway by SB203580 did not exert any effect on CTGF-induced ILK expression. Conclusion CTGF can induce the expression of ILK in time- and dose-dependent manner in PTEC, which is partially dependent on MEK/ERK1, 2 and PI3-K signaling pathways and independent on p38 MAPK signaling.

【Abstract】 Objective To evaluate the effects of exercise training combined with β1 adrenoceptor antisense gene therapy on blood pressure, renal function, preprorennin mRNA, β1 adrenoceptor mRNA and protein in Goldblatt hypertensive rats. Methods Liposome and β1-AS-ODNs were injected intravenously in rats with 2 kidney 1 clip (2K1C) hypertension. Renal function and blood pressure were measured. Levels of renal β1 adrenoceptor and preprorenin mRNA were tested by RT-PCR and the level of β1 adrenoceptor protein was tested by Western blotting. Results On the basis of the magnitude and duration of hypotension, exercise training combined with β1-AS-ODN decreased blood pressure by up to 41 mm Hg for 4 weeks. Compared with the 2K1C group, the levels of urine protein [(45.82±6.56 ) vs. (29.12±5.22) mg/L, P<0.01] and BUN [(13.10±2.62) vs. (9.05±1.84) mmol/L, P<0.05] were significantly decreased respectively and Ccr was significantly increased (Ｐ<0.01) in exercise training combined with β1-AS-ODN group. Exercise training combined with β1-AS-ODN significantly decreased the levels of preprorenin mRNA, myocardial β1 adrenoceptor mRNA and protein (P<0.05, P<0.05, P<0.05, respectively). Conclusion Exercise training combined with β1-AS-ODN can significantly ameliorate blood pressure and renal function in hypertensive rats, which is associated with the reducing levels of renal β1 adrenoceptor mRNA and protein. The inhibition of overactive β1-adrenoceptor expression probably occurs in transcription and post-transcriptional stages.

【Abstract】 Objective To investigate the effects of α-MSH on kidney inflammation and renal function of rhabdomyolysis-induced acute renal failure(ARF) by glycerol in rats. Methods Forty-eight Sprague-Dawley rats were divided into four groups: normal control group received 10 ml/kg saline by i.m.. ARF group rats were injected with 50% glycerol (10 ml/kg, i.m.). α-MSH immediate group and α-MSH delayed group were respectively given α-MSH (200 μg/kg, i.p.) immediately or 6 h afterward in addition to 50% glycerol injected(10 ml/kg, i.m.), and repeated after 12 h. Blood samples and kidney tissues were taken from anaesthetised rats at 24 h. Plasma creatine kinase (CK), BUN, Scr, IL-6 and TNF-α levels, as well as kidney acute tubular necrosis scores, ED-1 cell, and MCP-1 mRNA expression were investigated. Results Kidney tissues of ARF rats had more ED-1 positive cells （1.46±1.24 vs. 16.8±7.0, P<0.05）and higher MCP-1 mRNA expression (7.8±1.9 vs. 11.0±3.8, P<0.05) compared with normal control rats. α-MSH immediate group had less ED-1 positive cells(9.5±8.2) and lower MCP-1 mRNA expression(7.8±5.1) in kidney compared with ARF group(P<0.05). Also compared with ARF group, α-MSH immediate group had a lower BUN [(23.8±9.3) vs. (56.0±10.0) mmol/L, P<0.05], lower Scr [(152±76) vs. (333±60) μmol/L, P<0.05] and lower ATN score（1.7±0.4 vs. 2.7±0.4， P<0.05）. There were no significant differences of above parameters between α-MSH delayed group and ARF group. Conclusion There is obvious inflammation in kidneys of rhabdomyolysis-induced ARF rats, and α-MSH immediately administration can attenuate the inflammation and protect acute renal injury.

【Abstract】 Objective To investigate the influence of immunosuppressor mycophenolate mofetil(MMF) alone and in combination with valsartan (the angiotensinⅡ receptor blockers, ARB), on prevention of podocyte loss in streptozotocin(STZ) induced diabetic model. Methods Diabetes was induced in uninephrectomized male Wistar rats by peritoneal injection of STZ 65 mg/kg. Rats were randomly separated into five groups: control group(NC), diabetic without treatment group(DM), Valsartan treated group(V,40 mg&#8226;kg-1&#8226;d-1), MMF treated group(M,15 mg&#8226;kg-1&#8226;d-1), and combined treatment group(V+M). This study lasted for 8 weeks. Serum biochemistry, 24 h urinary protein and the ratio of kidney weight/body weight were determined after 8 weeks. The renal tissue morphology was observed by light microscopy and electron microscope. Expressions of nephrin, desmin and MCP-1 protein were examined by sem-quantitative immunohistochemical assays. Real-time quantitative PCR was used to detect the mRNA levels of nephrin and MCP-1 in renal tissue. Results Compared with NC, serum glucose level, 24 h urinary protein and the ratio of kidney weight/body weight in DM were significantly increased （P<0.01）. Treatment with either MMF or valsartan or combined treatment with valsartan and MMF significantly decreased 24 h urinary protein and the ratio of kidney weight/body weight, suppressed glomerulosclerosis and interstitial fibrotic lesions in diabetic rats. Nephrin mRNA expression in diabetic rats was lower than that of control rats(78%, P<0.05). Treatment with MMF in diabetic rats significantly restored nephrin mRNA expression. MCP-1 mRNA level in renal cortex was significantly higher in diabetic rats than that of control rats(251%, P<0.01).Over-expression of MCP-1 mRNA in diabetic rats was significantly suppressed by MMF(126%, P<0.01). There were significant negative correlations between nephrin and MCP-1 mRNA(r=-0.86, P<0.01), 24 h urinary protein and mRNA levels of MCP-1(r=0.56, P<0.01) or nephrin(r=-0.78, P<0.01). Conclusions MMF and valsartan can suppress MCP-1 and desmin expression, increase the nephrin expression, and decrease proteinuria level in diabetic rats. Combination of valsartan and MMF has no superiority over monotherapies on renal protection. These results suggest that MMF may have renoprotective effects on the early stages of diabetic nephropathy through an ant-inflammatory activity thus preventing podocytes loss.

【Abstract】　 Objective To investigate the effects of Rhizoma paridis pharmacologic serum on rat mesangial cell(MC) proliferation and apoptosis, and to explore the underlying mechanism. Methods Sera containing Rhizoma paridis(low, moderate and high dosages) and the positive control medicine such as multiglycosides Trioterygil wilfordii (MTW) and prednisone were prepared and put together with lipopolysaccharide (LPS). Rat MC was cultured in vitro for 12 h, 24 h and 36 h respectively. The effects of the drugs-containing sera on the proliferation of MC were observed by Cell Counting Kit-8(CCK-8) colorimetric assay. Apoptosis and apoptosis rate of MC were detected respectively by Hoechst 33258 fluorescence staining method and flow cytometry. The expression of bcl-2 mRNA of MC was evaluated by reverse transcription polymerase chain reaction (RT-PCR) method. Results Compared with LPS group, A amounts of Rhizoma paridis groups, MTW group and prednisone group were significantly decreased(P<0.01 or P<0.05). Rhizoma paridis pharmacologic serum inhibited MC proliferation in a dose-dependent and time-dependent manner. MC apoptosis was obviously induced by Rhizoma paridis pharmacologic serum, in a dose-dependent manner. Apoptosis rates of MTW group were significantly higher than those of all Rhizoma paridis groups. Prednisone pharmacologic serum did not induce MC apoptosis. Rhizoma paridis pharmacologic serum inhibited expression of MC bcl-2 mRNA(P<0.01). Compared with MTW, the inhibitory effect of Rhizoma paridis was less significant. Prednisone pharmacologic serum did not inhibit expression of MC bcl-2 mRNA. Conclusions Rhizoma paridis pharmacologic serum can partially inhibit LPS-induced MC proliferation and induce MC apoptosis. Rhizoma paridis pharmacologic serum can down-regulate the expression of bcl-2 mRNA, which partly explains the mechanism of Rhizoma paridis pharmacologic serum induced apoptosis. Rhizoma paridis can improve proliferative glomerulopathy possibly through inhibiting proliferation and promoting apoptosis.

【Abstract】 Objective To investigate the mechanism of erythropoietin（EPO）inhibiting the apoptosis of renal tubular epithelial cells stimulated by aristolochic acid（AA）. Methods LLC-PK1 cells were stimulated by different concentrations of AA（5, 10, 20 mg/L）and EPO（5, 10, 20 U/ml） was added as intervention. The media of the control contained neither of the agents. Serum concentration was uniform in all groups. In situ cells apoptosis were detected by TUNEL and the proportions of apoptosis were assessed by flow cytometry. The expression levels of caspase-3 and bcl-xL were assessed with immunoblotting after 24 h incubation. Results Compared with the control group, the expression of caspase-3 was increased slightly in AA 5 mg/L group and was increased remarkably in AA 10 mg/L group, whereas the expression was decreased in AA 20 mg/L group. EPO 10 U/ml and EPO 20 U/ml remarkably down-regulated the protein expression of caspase-3 induced by AA 10 mg/L. The expression of bcl-xL was increased remarkably in AA 5 mg/L group compared with the control group and the expression was decreased in AA 10 mg/L group, especially in AA 20 mg/L group. Compared with the AA 10 mg/L group, the expression of bcl-xL was increased in AA 10 mg/L+EPO 5 U/ml group and it was increased markedly in AA 10 mg/L+EPO 10 U/ml group. The expression in AA 10 mg/L+EPO 20 U/ml group was similar to that in AA 10 mg/L+EPO 10 U/ml group. Conclusion EPO can inhibit the excessive apoptosis of renal tubular cells stimulated by aristolochic acid through promoting the expression of bcl-xL and reducing the over-expression of caspase-3.