Hemin, a heme oxygenase-1 inducer, improves aortic

Background Under an insulin resistance （IR） state, overproduction of reactive oxygen species （ROS） may be playing a major role in the pathogenesis of endothelial dysfunction, hypertension and atherosclerosis. Recently, increasing attention has been drawn to the beneficial effects of heme oxygenase-1 （HO-1） in the cardiovascular system. This study aimed to investigate the effects of HO-1 on vascular function of thoracic aorta in IR rats and demonstrate the probable mechanisms of HO-1 against endothelial dysfunction in IR states.
Methods Sprague-Dawley （SD） rats fed with high-fat diet for 6 weeks and the IR models were validated with hyperinsulinemic-euglycemic clamp test. Then the IR rat models （n=-44） were further randomized into 3 subgroups, namely, the IR control group （n=26, in which 12 were sacrificed immediately and evaluated for all study measures）, a hemin treated IR group （n=-10） and a zinc protoporphyrin-Ⅸ （ZnPP-Ⅸ） treated IR group （n=-8） that were fed with a high-fat diet. Rats with standardized chow diet were used as the normal control group （n=-12）. The rats in IR control group, hemin treated IR group and ZnPP-Ⅸ treated IR group were subsequently treated every other day with an intraperitoneal injection of normal saline, hemin （inducer of HO-1, 30 μmol/kg） or ZnPP-Ⅸ （inhibitor of HO-1, 10 μmol/kg） for 4 weeks. Rats in the normal control group remained on a standardized chow diet and were treated with intraperitoneal injections of normal saline every other day for 4 weeks. Systolic arterial blood pressure （SABP） was measured by tail-cuffed microphotoelectric plethysmography. The blood carbon monoxide （CO） was measured by blood gas analysis. The levels of nitdc oxide （NO）, inducible nitric oxide synthase （iNOS）, endothelial nitric oxide synthase （eNOS）, blood glucose （BG）, insulin, total cholesterol （TC） and triglyceride （TG） in serum, and the levels of total antioxidant capacity （TAOC）, malondialdehyde （MD     

Erythroblast differentiation at spleen in Q137E mutant ribosomal protein S19 gene knock-in C57BL/6J mice

We recently found that erythroblast-like cells derived from human leukaemia K562 cells express C5a receptor (C5aR) and produce its antagonistic and agonistic ligand ribosomal protein S19 (RP S19) polymer, which is cross-linked between K122 and Q137 by tissue transglutaminases. RP S19 polymer binds to the reciprocal C5aRs on erythroblast-like cells and macrophage-like cells derived from human monocytic THP-1 cells and promotes differentiation into reticulocyte-like cells through enucleation in vitro. To examine the roles of RP S19 polymer in mouse erythropoiesis, we prepared Q137E mutant RP S19 gene knock-in C57BL/6J mice. In contrast to wild-type mice, erythroblast numbers at the preliminary stage (CD71^high/TER119^low) in spleen based on transferrin receptor (CD71) and glycophorin A (TER119) values and erythrocyte numbers in orbital artery bloods were not largely changed in knock-in mice. Conversely, erythroblast numbers at the early stage (CD71^high/TER119^high) were significantly decreased in spleen by knock-in mice. The reduction of early erythroblast numbers in spleen was enhanced by the phenylhydrazine-induced pernicious anemia model knock-in mice and was rescued by a functional analogue of RP S19 dimer S-tagged C5a/RP S19. These data indicated that RP S19 polymer plays the roles in the early erythroblast differentiation of C57BL/6J mouse spleen.     

血晶素促进缺氧大鼠肺组织诱导型血红素氧合酶基因的表达

目的　研究血晶素 (Hemin)对缺氧大鼠肺组织诱导型血红素氧合酶 (HO 1)表达的影响。方法　常压缺氧 15d复制缺氧性肺动脉高压大鼠模型 ,分为 3组 :正常对照组、单纯缺氧组和血晶素组。应用逆转录聚合酶链反应 (RT PCR)检测大鼠肺组织HO 1mRNA水平 ,双波长分光光度法检测动脉血中一氧化碳血红蛋白 (COHb)的含量 ,并用右心导管检测右心室收缩压。结果　缺氧 15d大鼠肺组织HO 1mR NA水平及血中COHb较正常大鼠升高 (P <0 0 1) ,同时右心室收缩压 (RVSP)升高 (P <0 0 1)。经血晶素干预后 ,使缺氧大鼠HO 1mRNA和COHb进一步增高 ,而RVSP降低(P <0 0 1或P <0 0 5 )。结论　血晶素能够上调HO 1基因在缺氧大鼠肺组织中的表达 ,促进内源性CO生成 ,抑制大鼠缺氧性肺动脉高压的形成     

血晶素对缺氧大鼠肺组织PDGF-B基因表达的影响

目的　研究血晶素对大鼠肺组织血小板源性生长因子B链 (PDGF B)基因表达的影响 ,探讨血晶素降低缺氧大鼠右心室收缩压 (RVSP)和改善缺氧性肺血管重构的机制。方法　用右心导管检测RVSP ;以双波长分光光度法测定动脉血中碳氧血红蛋白 (COHb)的含量 ;用免疫组织化学染色观察PDGF B、增殖细胞核抗原 (PCNA)蛋白的表达 ;用原位杂交检测PDGF BmRNA的定位并用图象分析检测其水平。结果　①正常大鼠肺腺泡内动脉 (IAPA)管壁PDGF BmR NA原位杂交及PDGF B、PCNA免疫组织化学染色结果为阴性 ,而缺氧大鼠的上述实验结果均为阳性 ,同时缺氧大鼠动脉血中COHb的含量较正常对照组大鼠高 1 8倍 (P <0 0 1)。②血晶素可使动脉血中COHb含量较单纯缺氧组进一步增多 (P <0 0 5 ) ,抑制缺氧大鼠RVSP的升高 (P <0 0 1) ,肺组织IAPA管壁PDGFmRNA原位杂交及PDGF B、PCNA免疫组织化学染色虽为阳性 ,但弱于单纯缺氧组 ,经图像分析差异有显著性 (P <0 0 1或 0 0 5 )。结论　血晶素促进内源性CO的产生 ,使缺氧大鼠肺组织PDGF B基因表达下调 ,间接地抑制缺氧状态下血管壁平滑肌细胞过度增殖引起的肺血管重构 ,这可能是血晶素改善大鼠缺氧性肺动脉高压的原因之一     

血红素加氧酶-1诱导对鼠肝缺血再灌注损伤的保护作用

目的研究血红素加氧酶-1(heme oxygenase-1,HO-1)在鼠肝缺血再灌注损伤肝组织中的表达及其作用。方法建立小鼠部分肝脏热缺血再灌注损伤模型,36只清洁级Balb/C小鼠随机分为3组: 假手术组(S组)、缺血/再灌注损伤组(I/R组)、HO-1诱导剂氯化高铁血红素(hemin)预处理组(HM组)。免疫组化半定量分析肝组织HO-1蛋白的表达,检测血清AST和ALT,肝组织丙二醛(MDA)含量及超氧化物歧化酶(SOD)活性,并观察肝组织的病理变化。结果与S组比较,I/R组HO-1蛋白表达显著增强,hemin预处理后,HO-1蛋白表达较I/R组增高(P<0．01)。I/R组AST,ALT活性和MDA的含量显著高于S组,而HM组均显著低于I/R组(P<0．01);I/R组SOD活性下降,而HM组显著高于I/R组(P<0．01)。HM组病理损伤程度明显轻于I/R组。结论 HO-1在鼠肝缺血再灌注损伤肝组织中表达上调,对肝脏具有保护效应。     

脑红蛋白对大鼠局灶性脑缺血组织的保护作用

目的探讨增加脑红蛋白（neuroglobin,Ngb）表达对大鼠在体局灶性脑缺血组织保护作用的影响。方法采用Hemin诱导脑红蛋白表达的方法,将SD大鼠随机分为5组,分别为假手术对照组、手术组、术后1h Hemin干预组、术后2h Hemin干预组、术后6h Hemin干预组。5组大鼠于术后24h检测脑梗死体积比,通过免疫组织化学及Western blot分析脑红蛋白表达情况。结果术后各时相点Hemin干预组脑梗死体积比较手术组有显著性差异（P〈0.05）;各干预组间比较,术后1h Hemin干预组脑梗死体积比较术后2h、术后6h Hemin干预组显著缩小（P〈0.05）。免疫组化分析,各时相点Hemin干预组Ngb阳性神经元数量较手术组增多,以术后1h Hemin干预组最明显。Western blot分析提示手术组、各时相点Hemin干预组Ngb表达均有非常显著升高（P〈0.01）。与手术组比较,术后1h Hemin干预组升高非常显著（P〈0.01）,术后2h Hemin干预组升高显著（P〈0.05）,术后6h组无明显差异（P〉0.05）。Ngb表达与脑梗死体积比呈负相关。结论大鼠急性局灶性脑缺血后增加脑红蛋白表达可以减轻缺血脑组织损伤。     

Effect of the heme oxygenase inducer hemin on blood haemostasis measured by high-frequency ultrasound

1. Heme compounds, like hemin, a heme oxygenase-1 inducer, are used in the treatment of acute porphyria treatment. The side-effects of hemin on haemostasis have been reported. To address those effects, in the present study we used a sensitive, high-frequency ultrasound technique to record acoustic velocity and to investigate whole blood clotting in Wistar rats treated chronically with hemin (50 mg/kg per day). 2. The hemin-induced disturbances in haemostasis measured were comparable to the heparin reference treatment, with a significant decrease in clotting velocity in both groups compared with controls (e.g. the time to clot was 40 +/- 5, 53 +/- 13 and 10 +/- 2 min, respectively; P < 0.05). Precautions must be taken when using high doses of hemin or in the treatment of bleeding diseases. 3. Further investigations are required to explore the effects of hemin in thrombosis models, because it could be a promising 'old drug' for the treatment of venous thrombosis in patients.     

兔急性肺损伤vWF变化及血红素加氧酶的保护作用

目的观察内毒素(ET)致兔急性肺损伤(ALI)时血浆血管性假血友病因子(vWF)含量变化及血红素加氧酶-1(HO-1)对其的影响。方法将24只雄性新西兰白兔随机分为对照组,内毒素组和血晶素组。对照组经颈静脉注射生理盐水;内毒素组经颈静脉注射内毒素(700μg/kg)复制ALI模型;血晶素组于手术前连续2d经腹腔注射血晶素(40μmol/kg,2次/d),其余操作同内毒素组。各组均于实验前(0h),实验后0.5、1、2、4h时点采动脉血行血气分析,ELISA方法检测血浆vWF值;处死动物后,测定肺组织干/湿重比,检测肺组织HO-1蛋白的表达情况,光镜下行肺组织病理学观察。结果与对照组比较,内毒素组静注ET后,PaO2、PaO2/FiO2下降,符合ALI诊断标准,0.5、1、2、4h时点血浆vWF明显升高(P<0.01),肺组织干/湿重比明显降低(P<0.01),病理学观察可见肺组织水肿、出血等改变;血晶素组肺组织HO-1平均光密度值明显高于对照组和内毒素组(P<0.01),4h时点血浆vWF值明显低于内毒素组(P<0.01),肺组织水肿、出血等改变较内毒素组有所减轻。结论ALI时血管内皮细胞受损,作为内皮细胞损伤标志物的血浆vWF含量升高,HO-1抑制内皮细胞受损,对ALI有一定的保护作用。     

Effect of Heme Oxygenase-1 Inducer Hemin on Chronic Renal Failure Rats

The role of HO-1 inducer, hemin, in chronic renal failure (CRF) rats and its possible mechanism of action was studied. 5/6 subtotal nephrectomy was performed to establish chronic renal failure model. Rats were randomly assigned to 4 groups: sham-operated group, CRF group,ferrous gluconate group and heroin group. At the 10th week after operation, serum creatinine,BUN, RBC, HGB and HCT were measured. Renal pathologic changes were observed. RT-PCR and immunohistochemistry were used to detect the expression and distribution of HO-1. RT-PCR and radioimmunoassay was used to determine the expression of ET-1 in the kidney and plasma. The results showed that as compared with CRF group, serum creatinine and BUN in hemin group were reduced significantly and nephrogenic anemia was improved markedly. Glomerular mesangial proliferation and interstitial lesion were also ameliorated significantly. Heroin not only increased the expression of HO-1 but also reduced the expression of ET-1 in the kidney. The level of ET-1 protein in the plasma was also reduced after heroin treatment. Most of these indexes were not obviously changed in ferrous gluconate group. It was suggested that through inducing the expression of HO-1 and reducing the level of ET 1 in the kidney and plasma, heroin plays an important protective role in 5/6 subtotal nephrectomized rats.