To explore the expression of heme oxygenase-1 (HO-1) in the peripheral blood mononuclear cells (PBMCs) and its relationship with pulmonary ventilation function in asthmatic patients, 18 asthmatic patients and 18 healthy subjects were selected. HO-1 protein and mRNA levels in PBMCs were measured by immunohistochemical staining and reverse transcription-polymerase chain reaction (RT-PCR), respectively. Blood carbon monoxide Hb (COHb), serum total IgE and pulmonary ventilatory function were observed. Our results showed that the percentage of cells positive for immunohistochemical staining of HO-1 were significantly higher in asthmatic patients (41.72 +/- 7.44) % than that in with healthy subjects (10.45 +/- 4.36) % (P < 0.001) and the optical density of PBMC HO-1 mRNA was higher in asthmatic patients (26.05 +/- 4.14) than that in healthy subjects (10. 82 +/- 4.26) (P < 0.001). The relation analysis showed that PBMC HO-1 protein and mRNA levels had significantly negative relation with FEV1%, PEFR, MEFR50%, respectively (r = -0.51-0.89, P < 0.05-0.001, respectively) and a positive relation with COHb and serum total IgE (r = 0.48-0. 85, 0.05-0.001, respectively). It is concluded that the expression of PBMC HO-1 protein and mRNA increased significantly in asthmatic patients, and HO-1 may play a significant role in the pathogenesis of asthma. The expression of HO-1 may bear a relation with severity of asthma.
Asthma/blood ; Asthma/*enzymology ; Carbon Monoxide/blood ; Heme Oxygenase-1/*biosynthesis ; Heme Oxygenase-1/blood ; Immunoglobulin E/*blood ; Leukocytes, Mononuclear/*enzymology ; RNA, Messenger/blood

AIMTo explore the expression of heme oxygenase-1 (HO-1) in the peripheral blood mononuclear cell (PBMC) and relationship to ventilatory function in asthmatic patients.

METHODSEighteen asthmatic patients and eighteen healthy subjects were selected. HO-1 protein levels in PBMC were measured by immunohistochemical staining and PBMC HO-1 mRNA were determined by reverse transcription-polymerase chain reaction (RT-PCR), blood carbon monoxide Hb (COHb) percent value, serum total IgE concentration and pulmonary ventilatory function were observed in asthmatic patients and healthy subjects.

RESULTSThe percentage of cells in immunohistochemical staining positive staining of HO-1 were significantly higher in asthmatic patients (41.7 +/- 7.44%) compared with that of healthy subjects (10.5 +/- 4.36%, P < 0.01), the optical densities of PBMC HO-1 mRNA were higher in asthmatic patients (26.05 +/- 4.14) compared with that of healthy subjects (10.82 +/- 4.26, P < 0.01). The relation analysis showed PBMC HO-1 protein levels had significantly negative relation with FEV1, PEFR, MEFR50%, respectively (r = -0.89, -0.56, -0.51, P < 0.01, respectively) and positive relation with COHb percent value, serum total I gE concentration (r = 0.80, 0.48, P < 0.05, respectively), and PBMC HO-1 mRNA levels had significantly negative relation with FEV1, PEFR, MEFR50%, respectively (r = -0.89, -0.65, -0.67, P < 0.05, respectively) and positive relation with COHb percent value, serum total IgE concentration (r = 0.85, 0.62, P < 0.01, respectively).

CONCLUSIONThe expression of PBMC HO-1 protein and mRNA are increased significantly in asthmatic patients, HO-1 may play a significant role in the pathogenesis of asthma. The expression of HO-1 has relation with severity of asthma.

Adult ; Asthma ; blood ; pathology ; Case-Control Studies ; Female ; Heme Oxygenase-1 ; blood ; Humans ; Leukocytes, Mononuclear ; metabolism ; Male ; Middle Aged

PURPOSE: Carbon monoxide (CO), a product of heme catalysis by heme oxygenase (HO-1, HO-2, HO-3), induces cytoprotection against ischemia/reperfusion (I/R) injury in a variety of organs such as the heart, lungs, kidneys, and liver. I examined whether CO would prevent chronic allograft nephropathy (CAN) associated with renal transplantation in rats. METHODS: Kidneys from male Fisher rats were perfused and harvested for transplantation. Lewis rats were used as recipients. After reperfusion of the implanted kidney, the recipient's remaining kidney was removed promptly. Recipients were then immediately treated with the indicated regimen of CO in a Plexiglas exposure chamber. At 90 days after transplantation, the animals were sacrificed for graft histopathology, serum creatinine, and blood urea nitrogen (BUN) as markers of kidney function. RESULTS: CAN in rats was achieved using a model of Fisher-to-Lewis transplants and evaluating kidney function over the 90 days following transplantation. CO administered at 100 ppm for 1 hr/day for 7 days prevented CAN at 90 days post-transplant. CO also decreased histopathological alterations, including leukocyte infiltration and cell death. CONCLUSION: These data expand our understanding of the protective effects of low-dose CO inhalation in preventing the development of chronic fibro-inflammatory changes associated with chronic allograft nephropathy and allow us to devise methods for improving long-term renal allograft function.
Animals ; Blood Urea Nitrogen ; Carbon ; Carbon Monoxide ; Catalysis ; Creatinine ; Cytoprotection ; Heart ; Heme ; Heme Oxygenase (Decyclizing) ; Heme Oxygenase-1 ; Humans ; Inhalation ; Kidney ; Kidney Transplantation ; Leukocytes ; Liver ; Lung ; Male ; Polymethyl Methacrylate ; Rats ; Reperfusion ; Transplantation, Homologous ; Transplants

Malaria is one of the most important public health problems in tropical areas on the globe. Several factors are associated with susceptibility to malaria and disease severity, including innate immunity such as blood group, hemoglobinopathy, and heme oxygenase-1 (HO-1) polymorphisms. This study was carried out to investigate association among ABO blood group, thalassemia types and HO-1 polymorphisms in malaria. The malarial blood samples were collected from patients along the Thai-Myanmar border. Determination of ABO blood group, thalassemia variants, and HO-1 polymorphisms were performed using agglutination test, low pressure liquid chromatography and polymerase chain reaction, respectively. Plasmodium vivax was the major infected malaria species in the study samples. Distribution of ABO blood type in the malaria-infected samples was similar to that in healthy subjects, of which blood type O being most prevalent. Association between blood group A and decreased risk of severe malaria was significant. Six thalassemia types (30%) were detected, i.e., hemoglobin E (HbE), β-thalassemia, α-thalassemia 1, α-thalassemia 2, HbE with α-thalassemia 2, and β-thalassemia with α-thalassemia 2. Malaria infected samples without thalassemia showed significantly higher risk to severe malaria. The prevalence of HO-1 polymorphisms, S/S, S/L and L/L were 25, 62, and 13%, respectively. Further study with larger sample size is required to confirm the impact of these 3 host genetic factors in malaria patients.
Agglutination Tests ; Blood Group Antigens ; Chromatography, Liquid ; Healthy Volunteers ; Heme Oxygenase (Decyclizing) ; Heme Oxygenase-1 ; Heme ; Hemoglobin E ; Hemoglobinopathies ; Hemoglobins ; Humans ; Immunity, Innate ; Malaria ; Plasmodium vivax ; Polymerase Chain Reaction ; Prevalence ; Public Health ; Sample Size ; Thalassemia

OBJECTIVETo study the impact of carbon monoxide (CO) on expression levels of inducible nitric oxide synthase (iNOS) mRNA in guinea pigs with allergic rhinitis (AR).

METHODSTwenty four guinea pigs were divided randomly into four study groups with 6 guinea pigs in each. The guinea pigs in the first group were treated with saline only (Group 1, the healthy controls). The remaing guinea pigs were sensitized by ovalbumin and thus establishing the AR models. After sensitization, the animals in the second group remained untreated (Group 2, AR control group). The third group was treated with Hemin as the induction group, and the fourth group was treated with Zinc protoporphyrin (ZnPP) as the suppression group. The plasma concentration of carboxyhemoglobin (COHb) was measured, which represents the concentration of CO. The expression levels of Heme oxygenase-1 (HO-1) and NOS mRNAs in nasal mucosa were determined by fluorescent quantitative RT-PCR.

RESULTSAR models were established successfully in all study guinea pigs. The concentrations of COHb (x(-) +/- s) in plasma of the second group (2.27% +/- 1.13%) were significantly (q = 4.10, P < 0.01) higher than those of healthy controls (1.08% +/- 0.24%). The plasma concentration of COHb in the third group (3.17% +/- 0.68%) were also significantly higher (q = 3.12, P < 0.05) than those in the second group. The expression levels of HO-1 and iNOS in nasal mucosa of the second group [(7.80 +/- 1.60) x 10(-3) and (5.81 +/- 0.05) x 10(-3), respectively] were also significantly (q equals 5.52 and 7.21, respectively, P < 0.01) higher than those of controls [(1.96 +/- 0.71) x 10(-3) and (0.97 +/- 0.05) x 10(-3), respectively]. The expression levels of HO-1 and iNOS in the nasal mucosa of the third group [(11.89 +/- 4.78) x 10(-3) and (7.42 +/- 0.70) x 10(-3), respectively] were significantly (q equals 3.86 and 2.22, P < 0.05) higher than those of the second group. The expression levels of HO-1 and iNOS in nasal mucosa of the fourth group [(3.82 +/- 0.98) x 10(-3) and (2.34 +/- 0.04) x 10(-3), respectively] were significantly (q equals 3.76 and 5.18, P < 0.05) lower than those in the second group.

CONCLUSIONSEndogenous carbon monoxide influenced the expression levels of iNOS in nasal mocusa in guinea pigs with AR.

Animals ; Carbon Monoxide ; blood ; Guinea Pigs ; Heme Oxygenase-1 ; Nitric Oxide Synthase Type II ; RNA, Messenger ; Rhinitis, Allergic

AIMTo explore the effect of ligustrazine injection on the expression of heme oxygenase-1 (HO-1) in rabbits with pulmonary ischemia/reperfusion injury after.

METHODSSingle lung ischemia/reperfusion injury animal model was used in vivo. Twenty rabbits were randomly divided into two groups( n = 10, in each), pulmonary ischemia/reperfusion injury (I/R) group and I/R + ligustrazine injection (LGT) group. The tissue slides were stained by immunohistochemistry (IHC) and in situ hybridization (ISH) for HO-1 to detect the expression of HO-1 in lung and to analyze the absorbance, wet to dry ratio of lung tissue weight (W/D) and the injured alveoli rate (IAR) were measured at 180 minutes after lung reperfusion. Meanwhile the lung tissue slide was prepared for electron microscopic observation at 180 minutes after reperfusion.

RESULTSHO-1 expression was upregulated in two groups in the pulmonary endothelial cells, part of pulmonary vascular smooth muscle cells, extima of vessels and epithelial cells of airway, the absorbance was 0.168 +/- 0.016 (0.148 +/- 0.013), 0.186 +/- 0.014 (0.158 +/- 0.012) respectively.The LGTI group showed higher absorbance than those of the I/R group (P < 0.01), lower W/D and IAR values than those of the I/R group (P < 0.01) significantly and lighter abnormal changes of the lung tissue in morphology than those of the I/R group.

CONCLUSIONLigustrazine injection possesses notable protective effects on I/R in rabbits by increasing the expression of HO-1 in lung.

Animals ; Disease Models, Animal ; Heme Oxygenase-1 ; metabolism ; Lung ; blood supply ; metabolism ; Pyrazines ; pharmacology ; Rabbits ; Reperfusion Injury ; metabolism

BACKGROUNDVascular smooth muscle cells (VSMCs) can express heme-oxygenase (HO), a rate-limiting enzyme in the degradation of heme to bilirubin, ferritin and carbon monoxide (CO). VSMC-derived CO can suppress VSMC proliferation and may serve as an antiproliferation factor. The promoter region of HO-1 shows a polymorphism with different (GT) n repeats that has been reported to differently induce gene expression. The objective of this study was to examine the effect of this variation on the occurrence of restenosis after in-stent treatment in patients with coronary artery disease.

METHODSCandidates who underwent coronary stent implantation were genotyped for the HO-1 promoter polymorphism using polymerase chain reaction (PCR) and automated DNA capillary sequencer. Serum levels of IL-6 and C-reactive protein (CRP) were obtained at baseline, 24 hours and 48 hours after stenting. The primary end point for the study was angiographic evidence of in-stent restenosis at 6 months. All parameters for evaluation of restenosis were analysed by quantitative computer-assisted angiographic analysis (QCA).

RESULTSOne hundred and eighty-seven patients who underwent coronary stent implantation were studied of whom 27.8% showed > or = 50% restenosis after 6 months. The distribution of (GT) n repeats of all patients in the promoter region of HO-1 genotype ranged from 22 to 42, with (GT) 25 and (GT) 32 being the two most common alleles. The allelic repeats were divided into the short class (S) with 29 (GT) n, the middle class (M) with 30-37 (GT) n and the long class (L) with 38 (GT) n. There was no significant difference in the restenosis between the genotype groups or between post operation levels of inflammation markers, but carriers of the S allele (n = 120) had 33.3% lower baseline IL-6 compared with non-S carriers (n = 67, P = 0.0008).

CONCLUSIONSAlthough no association was observed between the HO-1 promoter polymorphism and coronary in-stent restenosis following the stent procedure, the association with plasma IL-6 levels suggests that HO-1 S allele might protect from the atherosclerotic inflammatory process.

Adult ; Aged ; Angioplasty, Balloon, Coronary ; C-Reactive Protein ; analysis ; Coronary Restenosis ; blood ; enzymology ; genetics ; Female ; Genotype ; Heme Oxygenase (Decyclizing) ; genetics ; Heme Oxygenase-1 ; Humans ; Interleukin-6 ; blood ; Male ; Membrane Proteins ; Microsatellite Repeats ; Middle Aged ; Polymorphism, Genetic ; Promoter Regions, Genetic ; Stents

OBJECTIVETo study the effect of carbon monoxide (CO) on hydrogen sulfide (H2S) in guinea pigs with allergic rhinitis (AR) through intervention treatment.

METHODSAR model in guinea pigs was established by using ovalbumin. The animals were divided into three groups. Group one was sensitized continuously by ovalbumin, group two was treated with Hemin as induction group, and group three was treated with zinc protoporphyrin (ZnPP) as suppression group. The guinea pigs treated with saline were used as control. The behavior science scores, eotaxin concentration of nasal lavage, IgE in blood serum were recorded, and the plasma concentrations of CO and H2S were determined, then the expression of hemeoxygenase (HO)-1, cystathionine-beta-synthase (CBS) and cystathionine-gamma-lyase (CSE) were measured in nasal mucosa by fluorescent quantitative RT-PCR.

RESULTSThe behavior science scores, concentration of eotaxin in nasal lavage, IgE in blood serum and concentration of CO in plasma of sensitized group were higher than those of control (P<0.01), and the expression of HO-1 in nasal mucosa was also higher than control [(7.61+/-2.80)x10(-3) vs (2.32+/-1.14)x10(-3), P<0.05]. All these items were higher when treated with Hemin and lower when treated with ZnPP (P<0.05). The concentration of H2S in plasma was lower than control with significant differences [(14.80+/-1.60) micromol/L vs (18.90+/-1.00) micromol/L, P<0.01], the expression of CSE was also lower than control (P<0.05), and both of them were lower with Hemin induced and higher with ZnPP (P<0.05). The expression of CBS was very low and had no significant differences between groups (P>0.05), so it indicated that the CSE was the key enzyme for endogenous H2S product in nasal mucosa. Moreover the concentration of H2S was negatively correlated with CO (r=-0.702, P<0.001).

CONCLUSIONSEndogenous CO and H2S play a significant role in the pathogenesis of AR, and HO-1 and CSE are the main speed-relate enzymes respectively. The H2S is also influenced by CO.

Animals ; Carbon Monoxide ; blood ; Disease Models, Animal ; Guinea Pigs ; Heme Oxygenase-1 ; metabolism ; Hydrogen Sulfide ; blood ; Immunoglobulin E ; blood ; Male ; Rhinitis, Allergic, Perennial ; blood ; immunology

BACKGROUNDHeme-oxygenase 1 (HO-1) is a rate-limiting enzyme in the degradation of heme to bilirubin, ferritin and carbon monoxide (CO) and may have significant anti-inflammatory function. The HO-1 gene promoter region shows microsatellite polymorphism with different (GT)n repeats, reported to differently induce gene expression, with the short allele associated with higher gene expression. We measured the acute inflammatory response using coronary artery bypass surgery (CABG) as a well-characterized and uniform stimulus and examined the correlation between levels of IL-6, C-reactive protein (CRP) and fibrinogen and their relationship to HO-1 genotype.

METHODSTwo hundred and seventy-five consecutive patients undergoing CABG were genotyped for the HO-1 promoter polymorphism using PCR and automated DNA capillary sequencer. IL-6, CRP and fibrinogen were measured at baseline and 6, 24, 48, 72, 96 and 120 hours after CABG.

RESULTSComplete IL-6, CRP and fibrinogen measures were available in 220 patients. Before surgery IL-6 levels showed a strong correlation with CRP and fibrinogen (r = 0.48, P < 0.0001; r = 0.41, P < 0.0001 respectively), with a significant correlation between CRP and fibrinogen (r = 0.61, P < 0.0001). All three acute phase reactants showed a significant increase after CABG. After surgery, peak IL-6 was strongly correlated with peak CRP (r = 0.34, P = 0.0009) but not with peak fibrinogen (r = 0.15, P = 0.13), while peak CRP and peak fibrinogen were significantly correlated (r = 0.415, P < 0.0001). HO-1 allelic repeats ranged from 22-42, with (GT)25 and (GT)32 being the two most common alleles, and subsequently divided into three groups according to previous published work: <30 (GT)n were designated as S (short), 30-37 (GT)n as M (middle) and long repeats with >37 (GT)n as L (long); allele frequency 0.35, 0.58 and 0.07 respectively. Baseline CRP differed by genotype: those carrying at least one long allele having higher CRP than those with no long allele (3.76 +/- 0.79 vs. 2.07 +/- 0.17, P = 0.013). Conversely, those carrying at least one short allele had higher fibrinogen levels than those with no short allele (3.83 +/- 0.79 vs. 3.51 +/- 0.88, P = 0.006).

CONCLUSIONSThere is a strong correlation between the measured acute phase reactants both at baseline and after the inflammatory response to CABG in patients with coronary disease. There was an association between the HO-1 microsatellite polymorphism and CRP and fibrinogen levels at baseline but there was no similar association following CABG. This may indicate that HO-1 is associated with chronic atherosclerotic inflammatory processes rather than acute.

Adult ; Aged ; C-Reactive Protein ; analysis ; Coronary Artery Bypass ; Fibrinogen ; analysis ; Genotype ; Heme Oxygenase (Decyclizing) ; genetics ; Heme Oxygenase-1 ; Humans ; Inflammation ; etiology ; Interleukin-6 ; blood ; Membrane Proteins ; Microsatellite Repeats ; Middle Aged ; Polymorphism, Genetic

OBJECTIVETo explore the protein expression of heme oxygenase-1 (HO-1) and platelet endothelial cell adhesion molecules-1 (PECAM-1) in human coronary artery endothelial cells induced with Zinc Oxide Nanoparticle (ZnO-NPs).

METHODSMTT assay was used to determine the cell viability of ZnO-NPs. Levels of HO-1 and PECAM-1 protein in culture supernatants were measured using ELISA after human coronary artery endothelial cells were treated with different concentrations (0, 10, 20, 40µg/ml) of ZnO-NPs for 24 h.

RESULTSThe cell viability of human coronary artery endothelial cells in each group was 89.76%, 83.61%, 63.10%, 53.20%, 48.11%, 42.35%, 38.06%, 25.44% respectively when treated with different concentrations of ZnO-NPs (12.5, 25, 50, 70, 80, 90, 100, 200µg/ml). Protein levels of HO-1 (ng/L) in each group were 0.041±0.011, 0.512±0.076, 0.906±0.059, 1.062±0.089 respectively after the stimulation of different concentrations of ZnO-NPs (0, 10, 20, 40µg/ml). Comparisons in each group were statistically significant (P < 0.05). Protein levels of PECAM-1 (µg/L) in each group were 7.966 ± 0.046, 7.993 ± 0.036, 8.629 ± 0.052, 8.811 ± 0.039 respectively after the stimulation of different concentrations of ZnO-NPs (0, 10, 20, 40 µg/ml). Compared with the control group, protein levels of PECAM-1 increased (P < 0.05) when the concentration of ZnO-NPs was 20µg/ml or 40 µg/ml.

CONCLUSIONZnO-NPs stimulation could inhibit the viability of human coronary artery endothelial cells and upregulate the protein expression of HO-1 and PECAM-1.

Blood Platelets ; Cell Survival ; Coronary Vessels ; Endothelial Cells ; drug effects ; Heme Oxygenase-1 ; metabolism ; Humans ; Nanoparticles ; toxicity ; Platelet Endothelial Cell Adhesion Molecule-1 ; metabolism ; Zinc Oxide ; toxicity