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Objective:To explore the effect of electromyography biofeedback therapy combined with oxiracetam on peripheral blood heme oxidase-1 (HO-1), soluble apoptotic molecules and cognitive function in patients with senile vascular dementia (VaD).Methods:One hundred and fourteen elderly patients with VaD from May 2018 to May 2020 in Xingtai Third Hospital were selected and divided into two groups according to the random number table method, with 57 cases in each group. Both groups were given conventional treatment. On this basis, the control group was given oxiracetam, and the observation group was given electromyography biofeedback therapy combined with oxiracetam. The treatment effects after treated for 1 month was compared between the two groups. The levels of serum HO-1, soluble apoptotic molecules sFas, sFasL before and after treatment were compared between the two groups. Cognitive function evaluated by Wechsler Memory Scale (WMS), Mini Mental State Examination (MMSE). The scores of Chinese Stroke Scale (CSS), Ability of Daily Living (ADL) before and after treatment and adverse reactions were compared between the two groups.Results:After treated for 1 month, the total effective rate in the observation group was higher than that in the control group: 93.0%(53/57) vs. 77.2%(44/57), the difference was statistically significant ( P<0.05). After treated for 1 month, the level of serum HO-1 in the two groups was higher than that before treatment, and the level of serum HO-1 in the observation group was higher than that in the control group: (30.21 ± 4.05) μg/L vs. (24.19 ± 3.47) μg/L, the difference was statistically significant ( P<0.05). The levels of serum sFas and sFasL in two groups after treatment were lower than those before treatment, and the levels of serum sFas and sFasL in the observation group were lower than those the control group after treatment: (81.57 ± 16.23) ng/L vs. (118.49 ± 25.09) ng/L, (135.47 ± 24.41) ng/L vs. (200.71 ± 30.29) ng/L, the differences were statistically significant ( P<0.05). After treated for 1 month, the CSS scores in the observation group was lower than the control group: (13.48 ± 2.15) scores vs. (17.22 ± 3.02) scores; the WMS, MMSE, and ADL scores in the observation group were higher than those in the control group: (97.75 ± 10.27) scores vs. (88.43 ± 9.16) scores, (23.82 ± 2.50) scores vs. (21.38 ± 2.19) scores, (60.16 ± 6.24) scores vs. (51.29 ± 5.52) scores, the differences were statistically significant ( P<0.05). There was no significant difference in the incidence of adverse reactions between the two groups ( P>0.05). Conclusions:Electromyography biofeedback therapy combined with oxiracetam has a significant effect in the treatment of elderly patients with VaD. It can significantly improve vascular endothelial function, regulate apoptosis factors, strengthen cognitive function, promote recovery of nerve function and daily living ability without increasing adverse reactions.
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Objective To investigate the inhibitory effect oflentivirus-mediated polypyrimidine bundle binding protein-associated splicing factor (PSF) on retinal neovascularization (RNV) in mice model of oxygeninduced retinopathy (OIR).Methods One hundred and twelve 5-day-old C57BL/6J mice were randomly divided into normal control group,simple OIR model group,OIR model + lentivirus empty vector treatment group (Vec group) and OIR model + PSF lentivirus treatment group (PSF group),with 16,32,32 and 32 mice,respectively.When the mice were 7 days old,the mice in the normal control group were fed in a routine environment,and the mice in the OIR model group,Vec group and PSF group were established OIR model.The mice in the Vec group and PSF group were given an intravitreal injection of 1 μl of lentiviral vector and PSF lentivirus (titer 1 × 10~ TU/ml) at the age of 12 days.No injection was performed in the normal control group and simple OIR group.RNV was evaluated by counting the number of pre-retinal neovascular cells and analysis of non-perfusion area by immunofluorescent staining of the mouse retina.Real-time quantitative PCR was applied to detect the mRNA expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and hemeoxygenase1 (HO-1).Western blot analysis was applied to detect the protein expression ofNrf2,HO-1 and PSF.Results Of the normal control group,simple OIR model group,Vec group and PSF group,the number of pre-retinal neovascular cell nuclei were 0.00,14.36 ± 5.50,15.67 ± 4.96,8.13 ± 2.09,the non-perfusion area were 0.00％,(35.71 ± 2.81)％,(36.57 ± 4.53)％,(15.33 ± 4.75)％,respectively.The differences of the number of pre-retinal neovascular cell nuclei and non-perfusion area among 4 groups were significant (F=24.87,165.70;P＜0.05).Compared with the normal control group,there were more pre-retinal neovascular cell nucleis and larger nonperfusion area in the simple OIR model group and Vec group (P＜0.05).Compared with the simple OIR model group and Vec group,there were lower pre-retinal neovascular cell nucleis and smaller non-perfusion area in the PSF group (P＜0.05).Real-time quantitative PCR and Western blot showed that the mRNA expression of Nrf2,HO-1 (F=53.66,83.54) and protein expression ofNrf2,HO-1 and PSF (F=58.38,52.69,24.79) among 4 groups were significant (P＜0.05).The rnRNA expression ofNrf2,HO-1 and protein expression of Nrf2,HO-1 and PSF in the simple OIR model group and Vec group decreased significantly than those in the normal control group (P＜0.05).The mRNA expression ofNrf2,HO-1 and protein expression ofNrf2,HO-1 and PSF in the PSF group were increased significantly than those in the simple OIR model group and Vec group (P＜0.05).model group and Vec group (P＜0.05).Conclusion Intravitreal injection of lentivirus-mediated PSF inhibits RNV in mice model of OIR possibly through up-regulating the expression of Nrf2 and HO-1.
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Objective To observe the protective effect of polypyrimidine bundle-binding proteinrelated splicing factor (PSF) over-expression on RPE cell injury induced by advanced glycation end products (AGEs).Methods The human RPE cells cultured in vitro were divided into three groups:normal control group (N group),blank control group (N + AGEs group),empty vector control group (Vec + AGEs group),and PSF high expression group (PSF + AGEs).group).RPE cells in N group were routinely cultured;RPE cells in N + AGEs group were only transfected but did not introduce any exogenous genes combined with AGEs induction;Vec +AGEs group and PSF + AGEs group were transfected with pcDNA The empty vector or pcDNA-PSF eukaryotic expression plasmid was introduced into RPE cells and induced by AGEs.Except the N group,the other 3 groups of cells were transfected accordingly,and were stimulated with 150 μg/ml AGEs for 72 h after 24 h.HE staining and Hoechst 33258 staining were used to observe the effect of high PSF expression on the morphological changes of RPE cells;ROS level detection was used to analyze the effect of PSF high expression on the ROS expression of RPE cells induced by AGEs;MTT colorimetric method was used to detect the high PSF expression Effects on the viability of RPE cells;Western blot was used to detect the effects of different time and dose of PSF on the expression of heme oxygenase 1 (HO-1).Results HE staining and Hoechst 33258 staining observation showed that the cells in group N were full in shape,the nucleus was round,the cytoplasm was rich,and the staining was uniform;the cells in N + AGEs group and Vec + AGEs group were reduced in size,the eosinophilic staining was enhanced,and the nucleus was densely densely stained.Pyrolysis and even fragmentation;the morphology of cells in the PSF + AGEs group was still full,the cytoplasm staining was more uniform,and the nucleus staining was uniform.The results of MTT colorimetry showed that high expression of PSF can effectively improve the viability of RPE cells,but this effect can be effectively antagonized by ZnPP,and the difference is statistically significant (F=33.26,P＜0.05).DCFH-DA test results showed that compared with the N + AGEs group and Vec + AGEs group,the ROS production in PSF + AGEs group decreased,the difference was statistically significant (F=1 1.94,P＜0.05).Western blot analysis showed that PSF protein upregulated HO-1 expression in a time-and dose-dependent manner.The relative expression level of HO-1 at 24,48,and 72 h after PSF protein was significantly higher than that at 0 h,and the difference was statistically significant (F=164.91,P＜0.05).The relative expression level of HO-1 under the action of 0.1,0.5,1.0,1.5,and 2.0 μg PSF protein was significantly higher than 0.0 μg,and the difference was statistically significant (F=104.82,P＜0.05).Conclusion PSF may inhibit the production of ROS by up-regulating the expression of HO-1,thus protecting the RPE cells induced by AGEs.
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Objective To observe the effect oftert-Butylhydroquinone (tBHQ) on the expression of nuclear factor erythroid 2-related factor 2 (Nrf2),heme oxygenase (HO)-1 and phosphatidylinositol 3-kinase (PI3K) in high glucose cultured retinal Müller cells;and to investigate the anti-oxidative stress and anti-apoptotic effects oftBHQ.Methods Retinal Müller cells were divided into normal glucose group (5.5 mmol/L,N group),high glucose group (45 mmol/L,HG group) and tBHQ intervention group (HG+tBHQ group).After retinal Müller cells were cultured with high glucose for 48 hours,the pretreatment with tBHQ (20 μmol/L) induced the expressions of nuclear factor erythroid 2-related factor 2 (Nrf2) and HO-1.The Müller cells were identified by immunofluorescence staining.The expressions of Nrf2,HO-1,PI3K,B-cell lymphoma-2 (Bcl-2) and Bax were detected by Western blot and real-time fluorescence quantitative PCR.Flow cytometry was used to detect the apoptosis of retinal Müller cells in rats.Results Müller cytoplasm and nucleus GS showed strong positive,large cell body,abundant cytoplasm,uniform green fluorescence;nuclear DAPI staining round or oval,clear boundary.The expression of Nrf2 protein (t=4.114,P=0.006),HO-1 protein (t=9.275,P=0.000),Nrf2 mRNA (t=7.292,P=0.000) and HO-1 mRNA (t=15.014,P=0.000) in the HG group were higher than those in the N group.The expressions of Nrf2 protein (t=7.847,P=0.000),HO-1 protein (t=7.947,P=0.000),PI3K protein (t=5.397,P=0.002),Bcl-2 protein (t=6.825,P=0.000),Nrf2 mRNA (t=18.046,P=0.000),HO-1 mRNA (t=39.458,P=0.000),PI3K mRNA (t=4.979,P=0.003) and Bcl-2 mRNA (t=9.535,P=0.000) in the HG+tBHQ group were significantly higher than those in the HG group.The protein and mRNA expressions of Bax protein in the HG+tBHQ group were significantly lower than those in the HG group (t=14.998,16.520;P=0.000,0.000).Flow cytometry showed that the apoptosis rate of Müiller cells in the HG group was significantly higher than that in the N group (t=39.905,P=0.000).The apoptosis rate of Müller cells in the HG+tBHQ group was significantly lower than that in the HG group (t=21.083,P=0.000).Conclusion tBHQ can inhibit the apoptosis of retinal Müller cells by up-regulating the expression ofNrf2,HO-1 and PI3K.
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Objective To investigate the cellular viability and mitochondrial reactive oxygen species (ROS) production of the Müller cells under high glucose condition,and explore the protection role of the 5,6-dihydrocyclopenta-1,2-dithiole-3-thione (CPDT) on Müller cells.Methods Müller cells from Sprague Dawley rats were divided into 5 groups randomly,including 25 mmol/L normal glucose group (group A) and 65 mmol/L high glucose group (group B).High glucose group with 45,60,70 μmol/L CPDT and cultured them 72 hour was set as group C,D and E.Water soluble tetrazolium salt (WST)-8 was used to measure the cellular viability.Flow cytometry was used to measure the active oxygen and apoptosis index.The expression of nuclear factor erythroid 2-related factor 2 (Nrf2),hemeoxygenase-1 (HO-1),Bcl-2 and Bax protein were measured by Western blot.Results Compared with group A,the WST-8 showed that the viability of Müller cells apparently decreased in group B (t=39.59,P＜0.05).Compared with the group B,the viability of Müller cells had changes in group C (t=0.97,P＞0.05),but recovered in group D and E (t=-4.17,-7.52;P＜0.05).Compared with group A,the FCM showed that the mitochondrial ROS levels was higher in group B (t=-30.99,P＜0.05).Compared with group B,the mitochondrial ROS levels were decreased in group D (t=27.68,P＜0.05).Compared with group A,Bax,Nrf2 and HO-1 increased (t=-11.03,-63.17,-11.44;P＜0.05),while the bcl-2 decreased in group B (t=7.861,P＜0.05).Compared with the group B,Nrf2,HO-1 and Bax decreased (t=15.11,26.59,6.27;P＜0.05),while the bcl-2 increased in group D (t=-6.53,P＜0.05).Conclusions Under the high glucose,CPDT may reduce the mitochondrial ROS levels and the expression of Nrf2,HO-1 and Bax protein of Müller cells.It may inhibit apoptosis through activating the Nrf2/HO-1 pathway and balancing of level of Bcl-2 protein and mitochondrial ROS.
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Objective To compare the difference of serum heme oxygenase-1 (HO-1) and carbon monoxide(CO) levels between the patients with coronary heart disease(CHD) caused chronic heart failure(CHF) and CHD patients with normal cardiac function,and further to explore the protective mechanism of HO-1/CO system during the pathogenesis process of CHF.Methods Ninety-one patients with CHF were selected as the observation group and 72 CHD cases with normal cardiac function were taken as the control group.The concentration of HO-1 was determined by ELISA arid the Chalmer S method was used to detect serum CO concentration.The general clinical data of the two groups were recorded by the using the heart failure questionnaire.And the liver and kidney functions,blood lipids,NT-proBNP,BNP and cardiac echocardiography examination were performed.Results The serum HO-1 level in the observation group was (8.13±0.27)ng/mL,which was higher than (2.80±0.52)ng/mL in the control group;the CO level in the observation group was (0.35±0.06)mg/L,which was lower than(0.59±0.07)mg/L in the control group,the difference was statistically significant(P＜0.01);the HO-1 level in the observation group was gradually increased with the increase of cardiac function grade (P＜0.01);while the CO level was decreased with the increase of cardiac function grade (P＜0.01).Conclusion The serum HO-11evel in the patients with CHF is highly expressed with the heart failure aggravation;endogenous CO is gradually decreased due to consumption after cardiac failure aggravation.
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Objective To investigate the change of HO-1 expression in adipose tissue of obese young SD rats as well as its relationship with macrophage infiltration and polarization. Methods Three-week old SD rats (n=24) were randomly divided into 2 groups, routine diet group (NC) and high fat diet group (FC). After feeding 4 weeks, triglyceride (TG), high den?sity lipoprotein (HDL-C), fasting glucose and insulin were compared between these two groups and the insulin resistance in?dex was calculated. The gene expressions of HO-1, IL-6, IL-10 and MCP-1 were assessed by quantitative PCR. Infiltration and polarization of macrophages and M2 macrophages in the visceral adipose tissue were examined by immunohistochemis?try. Results The levels of FINS, FBG and HOMA-IR in rats of FC group were higher than those of rats in NC group after 4 weeks feeding (P0.05) in MOD value of F4/80 and CD206 between these two groups (P>0.05). Conclusion The infiltration of macrophage in visceral adipose tissue of obese young SD rats significantly increased while HO-1 expression was reactively increased. This insinuated that HO-1 might play an important role in anti-inflammatory mechanism through regulating polar?ization of macrophages.
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Objective To explore the expression of ininor histocompatibility antigen 13 (HM1 3) in gastric carcinoma and the relationship with clinicopathological parameters and prognosis.Methods The expression of HM 13 was detected by immunohistochemistry in a total of 90 pairs of paraffin-embedded gastric cancer tissue specimens and corresponding paraneoplastic tissues.The correlation between clinicopathological parameters and the expression of HM13 in gastric carcinoma were also analyzed.Downstream gene heme oxygenase-1 (HO-1) expression was detected by qRT-PCR after HM13 gene was interfered.Results High expression of HM13 protein was observed in 47％ gastric carcinoma compared with that in 61％ corresponding paraneoplastic tissues (x2 =3.78,P =0.052).HM13 expression has positive correlation with pathological TNM stage (x2 =5.022,P =0.025) and distant metastasis (P =0.033),but not with age (x2 =0.832,P =0.362),gender (x2 =0.779,P =0.378),tumor size (x2 =0.804,P =0.370),tumor differentiation (x2 =0.430,P =0.512),gross types (x2 =2.069,P =0.150),tumor stromal invasion (x2 =0.167,P =0.683) and lymph node status (x2 =0.396,P =0.529).The patients with low HM13 expression had worse overall survival [(OS):(30 ± 5) months vs.(47 ± 5) months,x2 =6.456,P =0.011] and progress free survival [(PFS):(29 ± 5) months vs.(46 ± 5) months,x2 =6.742,P =0.009].Expression of HO-1 was up-regulated after HM13 gene was interfered (0.532±0.013 vs.0.395±0.011,t=13.93,P＜0.05).Conclusion The low expression of HM13 is associated closely with advanced stage and distant metastases of gastric carcinoma.
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Objective To investigate the effects of heme oxygenase-1 (HO-1) transduced by cell penetrating peptide PEP-1 on renal ischemia/reperfusion (I/R) injury in rats.Methods Eighteen male Sprague-Dawley rats,aged 7-9 weeks,weighing 210-260 g,were randomly divided into 3 groups (n =6 each):sham operation group (group S),renal I/R injury group (group I/R) and fusion protein PEP-1/HO-I + I/R group (group HO).I/R injury was produced by occluding bilateral renal arteries for 45 min followed by reperfusion for 6 h.The fusion protein PEP-1/HO-1 was injected via the left iliac vein 30 min prior to ischemia in group HO.Bilateral renal arteries were only exposed but not occluded in group C.Blood samples were collected from the right common carotid artery at 6 h of reperfusion for determination of serum blood urea nitrogen (BUN) and creatinine (Cr) concentrations.The malondialdehyde (MDA) content,superoxide dismutase (SOD) activity and HO-1 expression in renal tissues were measured.Results Compared with group S,the levels of MDA,serum BUN and Cr were significantly increased,the SOD activity was decreased,and HO-1 expression was up-regulated in groups I/R and HO (P ＜0.05).Compared with group I/R,the levels of MDA,serum BUN and Cr were significantly decreased,the SOD activity was increased,and HO-1 expression was up-regulated in group HO (P ＜ 0.05).Conclusion HO-1 protein can be successfully transduced into renal tissues by PEP-1 and transduced HO-1 protein reduces renal I/R injury by inhibiting lipid peroxidation response.
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Objective To investigate the expression of heme oxygenase-1 (HO-1) in type 2 diabetes mellitus (T2DM) as well as hidden diabetes mellitus (HDM) and explore its clinical significance.Methods A total of 384 suspected patients with T2DM having underwent oral glucose tolerance test were enrolled in this study:216 cases of patients were T2DM (T2DM group),and 168 cases of patients were HDM (HDM group).T2DM patients were divided into single-vessel lesion group(122 cases),double-vessel lesion group(54 cases),and more-vessel lesion group(40 cases).Another 60 healthy person were as control group.The intima-media thickness (IMT) of carotid was measured by ultrasonography.The level of serum HO-1 was detected with enzyme-linked immunosorbent assay.Results The serum level of HO-1 in control group,HDM group and T2DM group was (1.24 ± 0.53),(2.12 ± 0.84),(3.46 ± 1.23) μ g/L.The serum level of HO-1 in T2DM group was significantly higher than that in control group and HDM group (P ＜ 0.05),and in HDM group it was significantly higher than that in control group(P ＜ 0.05).There were significant differences in the serum level of HO-1 and IMT of carotid among single-vessel lesion group,double-vessel lesion group,and more-vessel lesion group [(2.94 ± 1.14),(3.72 ± 1.36),(4.64 ± 1.58) μg/L and (1.21 ±0.16),(1.44 ± 0.20),(1.62 ± 0.27) mm] (P ＜ 0.05).The serum level of HO-1 and IMT of carotid in double-vessel lesion group was higher than that in single-vessel lesion group,and in more-vessel lesion group it was higher than that in double-vessel lesion group and single-vessel lesion group (P＜ 0.05 or ＜ 0.01).The serum level of HO-1 was positively related with IMT of carotid (r =0.512,P ＜0.01).Conclusion The high-level expression of HO-1 is found in HDM and T2DM patients,which maybe play a key role in the early diagnosis of T2DM.
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Objective To investigate the relationship between heme oxygenase-1 (HO-1),glutathione S-transferase (GST) and cerebral atherosclerosis.Methods Cerebralvascular status was assessed with color flow Doppler sonography,transcranial Doppler (TCD),magnetic resonance angiography (MRA)or/and digital subtraction angiography (DSA) in patients with cerebral atherosclerosis (mild,moderate,and severity).Serum HO-1 and GST were measured with enzyme-linked immunosorbent assay (ELISA).Results In comparison between case and control groups,there was significant difference in age,hypertension,cerebral infarction,uric acid,and HO-1 (P =0.041,0.008,0.000,0.036,and 0.001).The level of serum HO-1 in the severe atherosclerosis was lower than that in the mild and moderate atherosclerosis (P =0.000 and 0.002).Logistic regression was used to find the association of HO-1 and the degree of cerebral atherosclerosis (P =0.000).Conclusions HO-1 might be related to cerebral atherosclerosis.
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Gaseous molecules continue to hold new promise in molecular medicine as experimental and clinical therapeutics. The low molecular weight gas carbon monoxide (CO), and similar gaseous molecules (e.g., H2S, nitric oxide) have been implicated as potential inhalation therapies in inflammatory diseases. At high concentration, CO represents a toxic inhalation hazard, and is a common component of air pollution. CO is also produced endogenously as a product of heme degradation catalyzed by heme oxygenase enzymes. CO binds avidly to hemoglobin, causing hypoxemia and decreased oxygen delivery to tissues at high concentrations. At physiological concentrations, CO may have endogenous roles as a signal transduction molecule in the regulation of neural and vascular function and cellular homeostasis. CO has been demonstrated to act as an effective anti-inflammatory agent in preclinical animal models of inflammation, acute lung injury, sepsis, ischemia/reperfusion injury, and organ transplantation. Additional experimental indications for this gas include pulmonary fibrosis, pulmonary hypertension, metabolic diseases, and preeclampsia. The development of chemical CO releasing compounds constitutes a novel pharmaceutical approach to CO delivery with demonstrated effectiveness in sepsis models. Current and pending clinical evaluation will determine the usefulness of this gas as a therapeutic in human disease.
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Animals , Humans , Administration, Inhalation , Anti-Inflammatory Agents/administration & dosage , Carbon Monoxide/administration & dosage , Dose-Response Relationship, Drug , Environmental Pollutants/adverse effects , Gases , Heme/metabolism , Heme Oxygenase (Decyclizing)/metabolism , Inhalation Exposure/adverse effects , Risk Assessment , Signal TransductionABSTRACT
Cerebral vasospasm (CVS) is one of the serious complications of subarachnoid hemorrhage (SAH).Pathogenesis of CVS has not been fully clarified,and it may be associated with a variety of factors.With the development of molecular biology techniques,people have more understanding on SAH caused pathogenesis of CVS,and the research has also made considerable progress in the prevention of CVS.
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Objective To investigate the role of HO-1 in inhibition of oxygen-glucose deprivation (OGD)-induced apoptosis in rat hippocampal neurons by sevoflurane preconditioning.Methods Hippoeanlpal neurons of newborn Wistar rats (<48 h) were cultured in vitro.Tne neurons were randomly divided into 6 groups with 108 wells in each group:control group(group C),2% sevoflurane preconditioning group (group S1),OGD group,S1 +OGD group,4% sevoflurane preconditioning+OGD group (group S2+OGD),and 4% sevoflurane preconditioning+ZnPPⅨ+OGD group(group Z).Group C received no treatment.The neurons were cultured for 24 h after 2% sevoflurane preconditioning in group S1.For OGD experiments,the neurons were placed in deoxygenated glucose-free medium and sealed under 95% N2-5% CO2 in an anaerobic chamber equilibrated to 37℃ and 100%humidity for 45 min.then OGD was terminated by replacement of the stored medium and returning the cultures to a standard incubator maintained at 37℃ in 5% C02 and the neurons were cultured for 24 h as described by Ray et al. The OGD model was established after 2% and 4% sevoflurane preconditioning in group S1 + OGD and S2 + OGD respectively. In group Z, when the neurons were preconditioned with 4% sevoflurane, ZnPPⅨ was added to the culture medium at the same time, and the other procedures were the same as those in group S2 + OGD. The neuron viability, apoptesis rate, and expression of HO-I protein and mRNA were detected at 24 h of culture. Results Compared with group C, neuron viability was significantly decreased,apoptosis rate was significantly increased, and expression of HO-1 protein and mRNA was up-regulated in group OGD, S1 + OGD, S2 + OGD and Z, expression of HO-1 protein and mRNA was up-regulated in group S1 ( P < 0.01 ), but no significant change was found in neuron viability and apoptosis rate in group S1 ( P > 0.05). Compared with group OGD, neuron viability was significantly increased, apoptosis rate was significantly decreased, and expression of HO-1 protein and mRNA was up-regulated in group S1 + OGD and S2 + OGD ( P < 0.01), but no significant change was found in the indexes mentioned above in group Z ( P > 0.05 ). Neuron viability was significantly higher, apoptosis rate lower and expression of HO-1 protein and mRNA higher in group S2 + OGD than in group S1 + OGD ( P < 0.01). Neuron viability was significantly lower, apoptosis rate higher and expression of HO-1 protein and mRNA lower in group Z than in group S2+OGD(P<0.01).Conclusion HO-1 is involved in the inhibition of OGD-indueed apoptosis in rat hippocampal neurons by sevoflurane preconditioning.
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Objective To investigate the protective effects of heme oxygenase-1 (HO-1) mediated by cell penetrating peptide PEP-1 on myocardium against ischemia/reperfusion (IR) injury in isolated rat hearts. Methods Healthy male SD rats weighing 220-280 g were anesthetized with intraperitoneal pentobarbital. Their hearts were excised and perfused in a Langendorff apparatus with K-H solution saturated with 95%O2-5% CO2 at 37 ℃. Eighteen isolated rat hearts were randomly divided into 3 groups ( n = 6 each): Ⅰ group sham operation (group S);Ⅱ group IR and Ⅲ group PEP-1/HO-1 + IR (group HO-1). The isolated rat hearts were perfused with an oxygena-ted (95% O2-5% CO2 ) K-H solution at 37 ℃ in a Langendorff apparatus and were subjected to 40 min of global ischemia followed by 50 min of reperfusion after 30 min of stabilization. In group Ⅲ (group HO- 1 ) the isolated hearts were perfused with 50 μmol/L PEP-1/HO-1 for 15 min before ischemia. After 50 min of reperfusion, HO-1expression, MDA content and SOD activity in myocardial tissues were determined. The activities of creatine kinase (CK) and lactic dehydrogenase (LDH) in coronary effluent fluid were measured. Results The HO- 1 expression was significanfly higher in HO-1 group than in group IR. IR induced significant increase in MDA content and decrease in SOD activity in myocardium and CK and LDH activities in coronary effluent in group Ⅱ compared with group S. PEP-1/HO-1 significantly attenuated IR-induced changes. Conciusion HO-1 mediated by PEP-1 has protective effects on myocardium ngainst IR injury in rats.
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Objective To study the mechanism of monophosphoryl lipid A (MPLA) protecting liver ischemia-reperfusion injury in rats, and explore the hemeoxygenase-1-carbon monoxide-cyclic guanosine monophosphate (HO-1-CO-cGMP) pathway whether involved in MPLA enhance calcitonin gene-related peptides (CGRP) releasing or not. Methods Male SD rats were randomly divided into control group, sham-operated group, hepatic ischemia-reperfusion group, MPLA low, medium and high dose groups (hepatic ischemia-reperfusion + MPLA0. 2,0. 5,1.0 mg/ kg). Hepatic isehemia-reperfusion model was constructed, followed by observation of cell ultrastructure through electron microscope. Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH) and liver tissue levels of CO were determined. HO-1 expression in liver tissue was detected by immunohistochemic, CGRP, eGMP concentration in liver tissue was detected by RIA assay. Results Compared with hepatic isehemia-reperfusion group, the cell damage in MPLA group were relatively minor, and ALT, AST, LDH were significantly decreased (P <0.05), while HO-1, CO, cGMP, CGRP levels were signifi-cantly increased (P < 0.05). HO-1 and CO, CO and cGMP, cGMP and CGRP were obviously positive correlated (P <0.05). Conclu-sion MPLA enhanced CGRP synthesis and release through HO-1-CO-cGMP pathway in liver ischemia / reperfusion, which may be one of the mechanisms of MPLA reducing hepatic ischemia-reperfusion injury in rat.
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Objective To investigate the change rule and correlation of heme oxygenase-1(HO-1)/carbon monoxide(CO)and the influence of simvastatin and amlodipine in athemsclemtic progress.Methods The rabbits received 1％cholesterol diet(n=24)for eight weeks.After eight weeks,rabbits were fed with normal diet for eight weeks.The rabbits in model group(n=8)were administrated with cholesterol diet.The rabbits in simvastatin group(n=8)were administrated with simvastatin.The rabbits in amlodipine group(n=8)were administrated with amlodipine.The levels of serum lipids and plasma carbon monoxide were obtained at the beginning,the 8th week and the 16th week.The expression of heme oxygenase-1 in the thoraoia aortic tissue were observed with immunohistochemistry technique.Results By the end of 16th week,the levels ofserum lipids and plasma carbon monoxide in model group were obviously increased,however,the expression of heine oxygenase-1 were markedly decreased.Compared with model group.The levels～rurfl lipi&and plasma carbon monoxide in simvastatin group were significantly decreased,while the expression of heme oxysenase-1 in aortic great reduced.The levels flerum lipids in amlodipine group were not significant ckmged,the levels of plasma carbon monoxide were obviously decreased,while tlle expression ofheine oxygenase-1 in aortic great reduced.Conclusions In atheresclerofic progress,heme oxygenase-1(HO-1)/carbon monoxide(CO)appared the reciprocal relationship,and amlodipine may suppress athemsclemtie progress by decreasing the system.
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Objective To investigate the effects of peritoneal injection of cobaltic protoporphyrin Ⅸ chloride(CoPP)to induce heme oxygenase-1(Ho-1)upregulation in rat islet cells.Mthods Forty rats were divided into 5 groups by management 24 h before islets isolation:group A received inlzaporitoneal injection with 2.5 ms/ks CoPP,group B with 5 ms/ks CoPP,group C with 7.5 ms/kg CoPP,and group D with 10 mg/kg CoPP.In control group NS was used instead.A modified Goth approach was used for islet isolation.the yield and purity of the islets were assessed.The expression of Ho-1 mRNA and protein were detected by real time-PCR and Western blotting respectively.Islet function was tested by glucose stimulation test.Result Severe damage was found in tlle rats in group C and D.There was no difference in islet yield and purity for group A、B、C and control(P＞0.05).Group B had the highest Ho-1 mRNA and protein expression among the 4 groups.Though there was no difference in insulin secretion by low glucose challenge for group A、B and control's islets(P＞0.05),when challenged by hish level of glucose,significant deviation was observed.The imulin secretion level was(172.37±16.4)、(187.68±19.93)and(91.25±Conclusion Peritoneal iajection of 5 mg/kg CoPP can significantly enhance the expression of Ho-1 mRNA and protein in tat islet safely and enhance the function of islet when challenged by hish concentration of glucose.
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Objective To investigate the protein and mRNA expression of HO-1 in the lung tissue of asthmatic rat and the correlation between the percentage of blood carbon monoxide Hb(COHb)and the expressions of HO-1 in the lung tissue of asthmatic rat.Methods Twenty Sprague-Dawley(SD)male rats were randomly divided into 2 groups,control group and asthma group.Each group had 10 rats.The assessment of the percentage content of blood carbon monoxide Hb(COHb)wag performed.The total cell number and differentiation cell numbers in bronchoalveolar lavge fluid(BALF)and the inflammatory cells of airway wall were counted,the protein expressions of HO-1 in airway wall wag detected with immunohistochemistry technique.and the mRNA expressions of HO-1 in airway wall was detected with RTPCR.Results The expression of HO-1 was mainly located in airway epithelium and macrophage.The percentage of the cells in which protein of HO-1was expressed were(5.03±1.22)%,(27.14±4.68)%in two groups.The optical densities of mRNA expression of HO-1 were 0.323±0.05,0.68±0.02.The percent content of blood carbon monoxide Hb(COHb)were(0.45±0.35)%,(3.89±1.15)%.The protein and mRNA expressions of HO-1 and the percent content of COHb in asthmatic group were higher than those of the control group (P＜0.01 respectively).There was a significant positive correlation araong the protein expression of HO-1 in airway wall and the percent content of COHb(r=0.971,P＜0.001),mRNA expressions of HO-1 in lung tissue and the percent content of COI-Ib(r=0.897,P＜0.001).Conclusion The protein and mRNA expressions of HO-1 in the lung tissue,and the percent of COHb in blood were significantly mcreaged in asthmatic rat,which suggested HO-1 may play a significant role in the pathogenesis of asthma.
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Objective To clone the rat heme oxygenase-1(RHO-1) from rat spleen and express RHO-1 in E.coli BL-21.Methods The total RNA was extracted from rat spleen and amplified by reverse transcription polymerase chain reaction(RT-PCR).PCR products were cloned into pMD18-T(TA)vector followed by DNA sequencing.RHO-1 cDNA fragments in TA vector were subcloned into the prokaryotic expression vector pET28a(+).The recombinant pET28a(+)/RHO-1(rRHO-1) plasmid was transformed into E.coli.The rRHO-1 was induced with IPTG and characterized by SDS-PAGE.Results The cloned RHO-1 gene was composed of 870 nucleotides,and was accordance with the sequence reported in GenBank.The prokaryotic expression vector was constructed successfully.The RHO-1 protein was successfully expressed in E.coli.Conclusion The prokaryotic expression vector of rRHO-1 has been constructed,and the fusion protein has been successfully expressed.