ObjectiveThis study is designed to investigate the relationship between STR polymorphism in HO-1 gene promoter and susceptibility of CAD,in order to provide a new strategy for prevention and treatment of CAD by using HO-1.Methods200 patients who were diagnosed as CAD by coronary angiography were selected in this study.100 subjects without evidence of CAD under coronary angiography with their sex and age similar to CAD patients were selected as controls.Genotyping was performed using polymerase chain reaction followed by capillary electrophoresis automated DNA sequencer.Each size of the (GT) n repeat was calculated using the GeneMapper Analysis software.ResultsA (GT)n polymorphism was found in the HO-1 gene promoter with n =16 ～39.Subjects with n≤29 expressed much more HO-1 protein than those with n ＞29( P ＜0.01 ).The alleles were then classified into two subgroups,S'allele (n 29 ) and L'allele (n ＞29),the subjects were then classified as having an S/S,S/L,or L/L genotype.Subjects with the L allele ( L/L + L/S genotypes) had more chance to get CAD than those with S/S genotype ( adjusted OR =1.83,95 ％ CI =1.04 - 3.24).Stratified analysis further showed that L allele ( L/L + L/S genotypes) was susceptive to CAD in patients who smoke (adjusted OR =2.59,95％ CI =1.16 -5.80).ConclusionsThe (GT)n polymorphism in HO-1 gene promoter is related to susceptibility to CAD,especially in those patients who smoke.

Objective To investigate the relationship between hypoxia-inducible factor-1α(HIF-1α) and construction of collateral circulation in patients with coronary artery disease. Methods Collateral vessels were determined in 96 patients with≥70% narrowing of at least one coronary artery without prior revascularization,42 patients with coronary artery collaterals and 54 patients with no coronary artery collaterals.Another 50 cases with normal coronary arteries were selected as control.The levels of HIF-lα expression in monocytes and lymphocytes were tested by immunohistochemistry (IHC) and Western blot.Results Compared to the controls,the patients had higher expression of HIF-1α(P<0.01).Higher HIF-1α expression was found in patients with collaterals than in those without collaterals (P<0.01).Positive correlation was observed between the expression of HIF-1α protein and collateral score (r1=0.78,r2=0.84,P<0.01).Conclusion HIF-1α expression in circulating monocytes and lymphocytes are associated with collateral circulation.Detection HIF-1α might be helpful in the prognosis of patients with coronary artery disease.

Objective To observe the effects of statins, angiotensin-converting enzyme inhibitor/adrenergic receptor binder ( ACEI/ARB ) treatment on Heme Oxidase-1 ( HO-1 ) expression in patients with coronary artery disease. Methods HO-1 expression in periphery white blood cells was detected using Western blot before and after 3 months treatment of the above antioxidants in 60 patients who were diagnosed as coronary artery disease by coronary angiography. Results HO-1 expression measured by gray scale peak by western blot was significantly higher after 3 months antioxidant treatment than that before antioxidant treatment( 168. 9 t6. 8vs 112.7 ± 6.3, P = 0.008 ). Conclusion Effects of statins and ACEI/ARB in treating coronary artery disease may be due to its promoting expression of HO-1.

Objective To detect the signaling pathways involved in aldosterone (ALDO)induced mesangial cell (MC) proliferation. Methods The incorporation of 3H-thymidine (3H-TdR)and cell count were used as the measure of mesangial cell (MC) proliferation. Reactive oxygen species (ROS) production was determined by DCFDA fluorescence. Epidermal growth factor receptor (EGFR) activation was assayed by Western blotting. Results ALDO induced MC proliferation.When incubation with 100 nmol/L ALDO for 24 h, the 3H-TdR incorporation and cell number increased by 2.63- and 2.15-fold, respectively. Mineralocorticoid receptor (MR) antagonist EPLE almost completely blocked ALDO-induced MC proliferation (P<0.01), however, glucocorticoid receptor (GR) antagonist RU-486 had no effect on MC proliferation. ALDO increased intracellular ROS production in cultured human MCs. When incubation with ALDO (100 nmol/L) for 60 min,ROS production increased by 2.14-fold. ALDO-induced ROS generation was completely blocked by EPLE as well as mitochondrial complex Ⅰ inhibitor rotenone (P<0.01=, NADPH oxidase inhibitors diphenyleneiodonium sulfate (DPI) and apocynin inhibited ALDO-induced ROS production by 30%to 35% (P<0.05=. In contrast, inhibitors of other oxidant-producing enzymes, including allopurinol,indomethacin, nordihydroguiaretic acid, ketoconazole and G-nitro-L-arginine methyl ester (L-NAME)had no effect on ALDO-induced ROS production. Antioxidant N-acetyl-L-cysteine (NAC) and ROT inhibited ALDO-induced MC proliferation by 75% to 80%, whereas the inhibition of NADPH oxidase inhibitor apocynin and DPI on ALDO-induced MC proliferation was 25% to 30%. ALDO induced EGFR transactivation. When incubation with 100 nmol/L ALDO for 60 min, EGFR phosphorylation was increased by 4.95-fold, which was completely inhibited by EPLE and antioxidant NAC (P<0.01=. NAC and EGFR antagonist AG1478 significantly blocked ALDO-induced MC proliferation (P<0.01=. Conclusions ALDO-induced MC proliferation is mediated by ROS-dependent EGFR transactivation. ALDO-stimulated ROS is mainly generated by mitochondria.

AIM: To investigate the role of NF-?B/I?B signal pathway in the regulation of (cyclooxygenase-2) (COX-2) expression in human mesangial cells (HMC). METHODS: The PGE_2 concentration in supernatants of HMC was measured by radioimmunoassay. COX-2 mRNA and protein expression were determined by RT-PCR and Western blot. Electrophoretic mobility shift assay (EMSA) and Western blot were used to detect the activity of NF-?B and degradation of I?B. RESULTS: IL-1? significantly upregulated COX-2 expression and PGE_2 production in HMC. Significant up-regulation of NF-?B activation, nuclear translocation of p65 subunit, and degradation of I?B ? and I?B ? were observed in IL-1?-induced HMC. CONCLUSION: Expression of COX-2 in IL-1?-induced HMC is mediated by NF-?B/I?B signal pathway. [

Objective To investigate the role of oxidative stress-dependent Rasextracellular signal-regulated kinase (ERK1/2) signaling in aldosterone (ALDO)-induced mesangial cell proliferation. Methods The incorporation of 3H-thymidine (3H-TdR) and cell count were used as the measure of mesangial cell (MC) proliferation.Western blotting was used to detect the activation of Ki-RasA,c-Raf,MEK1/2,ERK1/2 and PI3K. Results Aldosterone significantly induced human mesangial cell proliferation,and anti-oxidant N-Acetylcysteine (NAC),catalase,and super oxide dismutase (SOD) significantly inhibited ALDO-induced mesangial cell proliferation (P＜0.01,respectively).Stimulation by ALDO for 3 h,Ki-RasA,c-Raf,MEK1/2,and ERK1/2 activity increased by 4.05-, 3.62-, 4.52-, and 3.40-fold compared with control group (P ＜0.01,respectively).NAC almost completely blocked ALDO-induced Ki-RasA,c-Raf,MEK1/2,and ERK1/2 activation (P＜0.01,respectively).Ki-RasA siRNA dose-dependently inhibited Ki-RasA expression, ALDO-induced Ki-RasA activation, and mesangial cell proliferation (P ＜0.01,respectively).c-Raf inhibitor GW5074 and MEK1/2 inhibitor PD98059 also reduced ALDO-induced mesangial cell proliferation by 65％ respectvely (P＜0.01).Ki-RasA siRNA had no effect on ALDO-induced PI3K phosphorylation.Combining LY294002 and PD98059 completely blocked ALDO-induced mesangial cell proliferation (P＜0.01). Conclusions ALDO-induced Ki-RasA-c-Raf-MEK-ERK signaling activation is dependent on reactive oxygen species (ROS) production,which mediates ALDO-induced mesangial cell proliferation.Inhibition of both ERK1/2 and PI3K signaling simultaneously completely blocks ALDO-induced mesangial cell proliferation.

Objective To elucidate whether Ang Ⅱ indnces the proliferation of mesangial cells through ROS-EGFR-JNK-AP-1 signaling pathway. Methods The incorporation of 3H-thymidine (3H-TdR) and cell count were used to measure mesangial cell (MC) proliferation. ROS production was determined by DCFDA fluorescence. EGFR and JNK activation was assayed by Western blot. Results Ang Ⅱ significantly enhanced ROS production in mesangial cells, which was up-regulated by 2.26 folds of control group after incubation with Ang Ⅱ for 60 min. Ang Ⅱ induced EGFR phosphorylation in dose- and time-dependent manner, with the peak (3.96 folds increase) at 30 min. EGFR phosphorylation was significantly blocked by AT1R antagonist losartan, antioxidant NAC, and NADPH oxidase inhibitor apocynin and DPI. EGFR antagonist AG1478 significantly inhibited Ang Ⅱ-induced mcsangial cell proliferation. Losartan, NAC, apocynin, DPI, and AG1478 ahnost abolished Ang Ⅱ-induced JNK activation. Conclusions ROS-EGFR-JNK-AP-1 signaling pathway is involved in Ang Ⅱ-induced mesangial cell proliferation. Apocynin and AG 1478 may be used as new therapy.

AIM: To investigate the role of CD2AP and F-actin in the pathogenesis of puromycin aminonucleoside nephrosis in rats. METHODS: Puromycin aminonucleoside nephrosis was induced by single intraperitoneal injection of puromycin aminonucleoside (PAN). Renal tissues were studied at 3, 7, 10 and 20 days after PAN injection by means of immunohistochemistry, RT-PCR, Western blotting and fluorescence. RESULTS: At day 3, CD2AP expression in podocytes began to decrease, and significantly decreased at day 7 and 10 (P

OBJECTIVE: To observe whether curcumin could inhibit the accumulation of the collagen IV and fibronectin in the glomeruli in nephrotoxi sera nephritis rats. METHODS: Seventy-two healthy male Sprague-Dawley rats were divided into three groups, with 24 animals in each group. For normal control group, normal saline (0.5 ml/d) was injected through intra-caudal-vein for two days, and at the same time normal saline (0.5 ml/kg) was also daily administered intraperitoneally. For nephrotoxic sera nephritis group, nephrotoxic sera (0.5 ml/d) was injected through the tail vein for two days and dimethyl sulfoxide (0.5 ml/kg) was given intraperitoneally daily. For curcumin group, nephrotoxic sera was injected as above and meanwhile curcumin (50 mg.kg(-1).d(-1)) was administered intraperitoneally every day. Six rats in each group were killed on the 3rd, 7th, 14th and 28th day. Their renal tissue was fixed in 10% formalin for examining the expression of collagen IV and fibronectin. RESULTS: Minimal staining of collagen IV and fibronectin was detected in the basement membrane of normal control rats glomeruli. In the nephrotoxic sera nephritis rats and curcumin treated nephrotoxic sera nephritis rats, the accumulation of collagen IV and fibronectin was increased progressively, with significant difference in the accumulation of collagen IV (P<0.01) between these two groups at the same time points, while the significant difference in fibronectin accumulation (P<0.05) appeared only after the 7th days. CONCLUSION: Curcumin can reduce the accumulation of collagen IV and fibronectin in the glomeruli. Hence we postulated that curcumin might have beneficial effect for retarding glomerulosclerosis.

Objective To explore the effects and mechanism of hydrogen sulfide on myocardial ischemia reperfusion in rats. Methods With sodium hydrogen sulfide (NaHS) as a donor of hydrogen sulfide ( H2S), we established myocardial ischemia-reperfusion injury model in rats. The SD rats were randomly divided into control group,myocardial ischemia reperfusion group (I/R group), H2S group,and H2S and glibenclamide (H2S + GLI)group. We monitored the hemodynamics index of rats, including heart rate, arterial pressure, left ventricular pressure. The rate of ventrical arrhythmia was also observed in each group. Results H2 S significantly reduced the ventricular arrhythmia (VA) occurrence (H2S group 66.5% vs I/R group 33.5% (P ＜0.05) and score in myocardial ischemia reperfusion rats (H2S group 2. 6 ±0. 7 vs I/R group 4. 5 ±0. 8(P＜0.05). The KATP channel blocker,glibenclamide,could weaken the antiarrhythmic effects of H2S ( H2S group 2. 6 ±0. 7 vs. H2S + GLI group 4. 0 ± 0. 6, P ＜ 0.05 ). Conclusions H2S has the protective effect against myocardial ischemia reperfusion damage. This function may be associated with the KATP signal transduction pathway in cells.