血红素加氧酶-1的抗肿瘤细胞凋亡作用研究进展

血红素加氧酶-1(heme oxygenase-1,HO-1)是血红素分解的关键酶,其促进血红素分解释放胆绿素、CO和亚铁离子。HO-1在机体内发挥多种生物学功能,主要有减轻炎症反应、降低氧化损伤、抑制细胞凋亡以及调节细胞增殖。随着研究的不断深入,HO-1与肿瘤的相关性研究逐渐增多,表明其与肿瘤关系密切,参与调控肿瘤细胞凋亡、增殖等过程。本文主要就HO-1蛋白与肿瘤细胞凋亡的相关性作一综述。     

血红素氧合酶-1与心血管系统的相关研究

血红素氧合酶-1(NO-1)是细胞受到外界刺激后诱导性表达的一种分解血红素的关键酶,其通过激活p55/肿瘤坏死因子受体-1(Tumor Necrosis Factor Receptor-1,TNFR-1)、p38丝裂原活化激酶(Mitogen-Activated Protein Kinases,MAPK)和磷脂酰肌醇-3激酶/蛋白激酶B(Phosphatidylinositol 3Kinase/Protein Kinase B,PI3K/AKt)等信号通路,抑制多种凋亡相关分子活性,发挥抗氧化、抗炎、抗凋亡、抗血栓与降压等作用,从而保护心血管系统,HO-1基因变异可能引起心血管疾病。     

HO-1与HCV的相关性研究

血红素加氧酶-1(heme oxygenase-1,HO-1)在肝脏和脾脏高表达,可以被包括某些病毒感染在内的多种因素诱导表达,具有抗氧化、抗炎、抗凋亡等保护作用.丙型肝炎病毒(hepatitis C virus,HCV)感染可以造成慢璎肝炎、肝硬化和肝癌等疾病,对人类健康造成很大威胁.研究发现HO-1通过影响HCV的复制发挥其保护作用,同时HCV也可以反向调控HO-1的表达.尽管HO-1与HCV相互作用的分子机制还不明确,但HO-1与HCV感染相关性研究不断取得重大进展,将为HCV感染的治疗提供一种新方法.     

诱导血红素加氧酶-1表达在器官移植中的保护作用

器官移植术中及术后移植器官的缺血再灌注损伤(ischemia-repeffusion injury,IRI)和免疫排斥反应一直困扰着外科医生.血红素加氧酶-1(heme oxygenase-1,HO-1)是血红素代谢过程中的限速酶,广泛分布于哺乳动物的各种组织细胞中.血红素在它的催化下降解代谢为一氧化碳(CO)、胆绿素和游离铁离子.HO-1在氧化应激、炎性反应、低氧和缺血等状态下均能高度表达.HO-1及其催化血红素代谢产物主要通过抗炎性反应、抗氧化反应、调节同种异体反应性T细胞的活性及增殖、抗内皮细胞凋亡、抑制内皮细胞活化等作用机制,对移植器官起到抗IRI和抗免疫排斥作用,从而增加移植器官成活率及延长其存活时间.     

血红素加氧酶-1生物学特性及在心肌梗死中的作用

血红素加氧酶（Heme Oxygenase，HO）是血红素降解过程中的限速酶，其在体内有三种形式：HO-1、HO-2、HO-3。近年来许多研究表明HO-1具有抗炎、抗凋亡等作用以及在心脏缺血／再灌注损伤等应激反应中起着重要的保护作用^[1,2]。     

血红素加氧酶和动脉粥样硬化关系的研究进展

血红素加氧酶(HO)是一种降解血红素的限速酶,它能催化降解血红素生成等摩尔质量的一氧化碳(CO),胆红素和游离铁离子。大量证据表明诱导型HO(HO-1)和其产物具有抗氧化损伤的功能.其具有细胞保护作用,机制可能是通过其降解产生的胆绿素及其随后胆红素的产生而发挥抗氧化剂的特性有关,其产生的CO,具有抗炎、抗增生和血管扩张作用。本综述将重点讨论HO与心血管疾病的关系,特别是粥样动脉硬化。HO-1及其代谢产物作用的研究将为心血管疾病的临床治疗开辟新的途径。     

1,6-二磷酸果糖减轻STZ致大鼠胰岛细胞凋亡

目的 探讨1,6-二磷酸果糖（FDP）对链脲佐菌素（STZ）致胰岛细胞凋亡的保护作用与血红素加氧酶/一氧化碳（HO-1/CO）、抗氧化以及NOS/NO系统的关系。 方法 用离体培养乳鼠胰岛细胞,分别检测STZ、FDP作用后细胞形态、细胞活性、细胞凋亡率、细胞HO-1的活性和细胞培养上清液中胰岛素的基础和高糖刺激分泌量以及CO的含量、超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶（GSH-PX）、诱生型一氧化氮合酶（iNOS）和NO的变化。结果：STZ作用后,胰岛细胞活性降低,基础和高糖刺激胰岛素分泌减少,胰岛细胞凋亡率明显增加（p＜0.01）;细胞HO-1活性有所下降,上清液中CO生成减少（p＜0.01）,SOD、GSH-PX活性水平降低、iNOS增多、NO增加。与FDP孵育后细胞活性显著升高,胰岛素基础和高糖分泌量增多,胰岛凋亡率明显降低, HO-1活性和CO生成明显提高（p＜0.01或p＜0.05,同时 SOD、GSH-PX活性水平明显增强、iNOS活性降低、NO生成减少,并且具有剂量依赖性。 结论 FDP能显著改善STZ致胰岛细胞凋亡,其机制可能与增加HO-1/CO系统水平 ,提高抗氧化酶SOD、GSH-PX的活性以及降低iNOS活性,从而减少NO的生成等途径有关。     

二十二碳六烯酸抑制氧化应激状态下人视网膜色素上皮细胞凋亡

 目的: 观察二十二碳六烯酸(docosahexaenoic acid,DHA)对外源性H₂O₂诱导人视网膜色素上皮细胞凋亡的影响及分子机制。方法: 体外培养人视网膜色素上皮细胞系ARPE-19,加入终浓度为12.5 mol/L的H₂O₂诱导氧化应激,随后用30~100μmol/L DHA作用细胞4~24 h;real-time PCR和Western blot分别检测血红素氧合酶-1(heme oxygenase-1,HO-1) mRNA和蛋白的表达;比色法分析HO-1酶活性;荧光探针检测活性氧簇(reactive oxygen species,ROS)的产生;免疫荧光检测转录因子NF-E2相关因子2(NF-E2-related factor 2,Nrf2)的核转位。最后通过HO-1 siRNA干扰后,流式细胞术观察其对ARPE-19细胞凋亡的影响。结果: DHA能以浓度依赖性方式诱导ARPE-19细胞表达HO-1 mRNA和蛋白,同时,HO-1的酶活性也随着DHA浓度的递增而增强;DHA处理也能诱导Nrf2核转位。此外,H₂O₂处理可促进ARPE-19细胞凋亡,并诱导其产生ROS。同时给予100μmol/L DHA处理后,细胞凋亡率和ROS生成显著降低。转染HO-1 siRNA或用HO-1抑制剂ZnPP处理后,可明显降低DHA对细胞凋亡率和ROS的抑制作用。结论: DHA可能通过Nrf2途径诱导视网膜色素上皮细胞表达HO-1,从而发挥对细胞的保护作用。     

嵌合抗原受体T细胞与病毒相关的应用进展

嵌合抗原受体T细胞(chimeric antigen receptor T cell, CAR-T)疗法是近年来新兴的细胞免疫疗法，它通过基因工程技术使T细胞表达特异性嵌合抗原受体(chimeric antigen receptor, CAR),以非MHC依赖形式识别并杀伤表达相应抗原的靶细胞。CAR-T疗法在血液系统恶性肿瘤的治疗中取得了令人振奋的结果，但在实体瘤治疗领域仍然存在很多困难。溶瘤病毒可直接裂解实体瘤细胞，逆转肿瘤局部的免疫抑制微环境，与CAR-T疗法联用可以增强抗实体瘤效果。此外，CAR-T疗法也显示出一定的抗病毒作用，主要表现在抗人类免疫缺陷病毒(human immunodeficiency virus, HIV)、HBV等方面。文章从CAR-T与病毒的关系入手，主要就CAR-T疗法抗病毒的研究现状及溶瘤病毒与CAR-T疗法联合抗实体瘤机制等方面展开综述，以期为拓宽CAR-T抗病毒治疗领域以及CAR-T的联合治疗方案制定提供新思路。     

点地梅提取物saxifragifolinB体外抗实体瘤活性研究

目的: 初步探讨点地梅提取物saxifragifolin B体外抗实体瘤活性及其作用机制。方法: Saxifragifolin B作用于体外培养的人肝癌细胞BEL-7402、肺癌细胞A549和胃癌细胞SGC7901。光镜下观察肿瘤细胞形态学变化;AO/EB双染法观察细胞死亡;MTT法检测细胞增殖水平,计算IC₅₀;流式细胞术检测细胞凋亡和坏死。结果: Saxifragifolin B诱导3种实体瘤细胞出现凋亡样和坏死样改变。Saxifragifolin B以浓度依赖的方式抑制肿瘤生长,作用24 h IC₅₀分别为13.13 μmol/L、9.61 μmol/L和11.64 μmol/L。Saxifragifolin B可以诱导肿瘤细胞凋亡和坏死。结论: 点地梅提取物saxifragifolin B具有抗多种实体瘤的活性,其作用机制可能与诱导细胞凋亡和促进细胞坏死有关。     

Heme oxygenase-1 in tumors: is it a false friend?

Heme oxygenase-1 (HO-1) catalyzes the oxidation of heme to biologically active products: carbon monoxide (CO), biliverdin, and ferrous iron. It participates in maintaining cellular homeostasis and plays an important protective role in the tissues by reducing oxidative injury, attenuating the inflammatory response, inhibiting cell apoptosis, and regulating cell proliferation. HO-1 is also an important proangiogenic mediator. Most studies have focused on the role of HO-1 in cardiovascular diseases, in which its significant, beneficial activity is well recognized. A growing body of evidence indicates, however, that HO-1 activation may play a role in carcinogenesis and can potently influence the growth and metastasis of tumors. HO-1 is very often upregulated in tumor tissues, and its expression is further increased in response to therapies. Although the exact effect can be tissue specific, HO-1 can be regarded as an enzyme facilitating tumor progression. Accordingly, inhibition of HO-1 can be suggested as a potential therapeutic approach sensitizing tumors to radiation, chemotherapy, or photodynamic therapy.     

Heme oxygenase-1 has antitumoral effects in colorectal cancer: Involvement of p53

The expression of heme oxygenase-1 (HO-1) has been shown to be up-regulated in colorectal cancer (CRC), but the role it plays in this cancer type has not yet been addressed. The aims of this study have been to analyze HO-1 expression in human invasive CRC, evaluate its correlation with clinical and histo-pathological parameters and to investigate the mechanisms through which the enzyme influences tumor progression. We confirmed that HO-1 was over-expressed in human invasive CRC and found that the expression of the enzyme was associated with a longer overall survival time. In addition, we observed in a chemically-induced CRC animal model that total and nuclear HO-1 expression increases with tumor progression. Our investigation of the mechanisms involved in HO-1 action in CRC demonstrates that the protein reduces cell viability through induction of cell cycle arrest and apoptosis and, importantly, that a functional p53 tumor suppressor protein is required for these effects. This reduction in cell viability is accompanied by modulation of the levels of p21, p27, and cyclin D1 and by modulation of Akt and PKC pathways. Altogether, our results demonstrate an antitumoral role of HO-1 and points to the importance of p53 status in this antitumor activity.     

小剂量丙泊酚对食管鳞癌细胞Eca109生物学行为的影响

目的:探讨低剂量丙泊酚对食管鳞癌细胞Eca109的增殖、凋亡、迁移及侵袭等生物学行为的影响。方法:根据预实验结果设置低剂量丙泊酚组,干预食管鳞癌细胞Eca109后,利用MTT、流式细胞术、transwell检测对增殖、凋亡、迁移及侵袭的影响,应用实时荧光定量PCR检测血红素氧合酶1(HO-1)的表达变化。结果:低剂量Eca109可以促进Eca109的增殖、迁移与侵袭并抑制凋亡,实时荧光定量PCR显示干预后HO-1的表达增加且随浓度的增加而增加。结论:低剂量丙泊酚可以促进Eca109的增殖、迁移与侵袭并抑制凋亡,可能与促进HO-1的表达相关。     

Probucol抑制氧化低密度脂蛋白诱导的大鼠主动脉平滑肌细胞增殖

目的:研究Probucol抑制ox-LDL诱导RASMCs增殖与信号蛋白分子ERK1/2、MKP-1、HO-1和Trx-1表达之间的关系。方法:采用MTT、流式细胞术和Western blotting观察ox-LDL刺激条件下probucol对细胞周期、细胞增殖和凋亡、ERK1/2、MKP-1、HO-1和Trx-1表达的影响。结果:(1)Probucol抑制ox-LDL刺激RASMCs增殖:100μmol/Lprobucol+35mg/Lox-LDL组与35mg/Lox-LDL组比较,A值下降了34.9%(P0.01);(2)Probucol通过使RASMCs停滞在G0/G1期和诱导细胞凋亡2种方式抑制ox-LDL刺激细胞增殖。(3)ox-LDL显著抑制MKP-1的蛋白表达,与对照组比较下降了60.0%(P0.01),同时使p-ERK1/2表达增加了34.7%;Probucol使MKP-1蛋白表达显著增加2倍,p-ERK1/2表达降低了15.7%(P0.01);(4)35mg/Lox-LDL使细胞内Trx-1蛋白表达下降28.9%(P0.05),HO-1蛋白表达轻度增加(P0.05)。与ox-LDL组比较,probucol使Trx-1蛋白表达增加了91.6%(P0.01),HO-1表达增加31.9%(P0.01)。结论:Probucol通过增强MKP-1和HO-1蛋白表达、抑制细胞周期运转和诱导细胞凋亡的机制抑制RASMCs增殖。     

Heme-induced heme oxygenase-1 (HO-1) in human monocytes inhibits apoptosis despite caspase-3 up-regulation

Monocyte activation, apoptosis and differentiation are hallmarks of most inflammatory vascular disorders. We studied the effects of heme oxygenase-1 (HO-1) induced by its substrate hemin on apoptosis, caspase-3 expression and the differentiation of freshly isolated human monocytes. Hemin induced HO-1 in a dose- and time-dependent fashion as measured by semi-quantitative RT-PCR and flow cytometry. Apoptosis was markedly suppressed by hemin in cells rendered apoptotic by serum deprivation or dexamethasone as determined by flow cytometric detection of annexin V binding or transmission electron microscopy (TEM). The specific HO-1 inhibitor zinc protoporphyrin (ZnPP) reversed the effects of hemin on monocyte apoptosis and diminished cell lifespan. Surprisingly, the cytoprotective effects of hemin were positively correlated with caspase-3 up-regulation. Hemin-induced apoptosis suppression was enhanced by the caspase-3 inhibitor DEVD-CHO, indicating that caspase-3 was active in a pro-apoptotic fashion. Hemin inhibited CD95 as a putative cytoprotective mechanism. Morphological studies and detection of CD86 showed that monocytes differentiated into macrophages in response to hemin after relatively long incubation times, a phenomenon that might be provoked by caspase-3-regulated pathways. Our results confirm a similar cytoprotective effect of hemin/HO-1 for monocytes as has been shown for other cells, despite caspase-3 up-regulation. The fact that HO-1 may adversely affect monocyte survival and differentiation could be of particular significance in future therapies for occlusive vascular diseases or transplant rejection.     

Significance of heme oxygenase in prolactin-mediated cell proliferation and angiogenesis in human endothelial cells

Our objectives were to determine whether heme oxygenase-1 is a second messenger for prolactin-mediated angiogenesis. Endothelial cell proliferation and angiogenesis assay demonstrated that cell number and capillary formation were increased by prolactin (10 and 25 ng/ml). Both protein synthesis and mRNA analysis confirmed that HO-1 expression was induced by prolactin in cultured endothelial cells and occurred in a concentration-dependent manner. Endothelial cells transduced with retrovirus-mediated delivery of HO-1 gene in sense and antisense orientation were used to further determine whether HO-1 overexpression or underexpression modulated prolactin-mediated endothelial cell proliferation and angiogenesis. Incubation of human microvessel endothelial cells transduced with HO-1 in sense orientation resulted in enhancement of prolactin-mediated increase in endothelial cell proliferation and angiogenesis, whereas inhibition of HO-1 by transduction of HO-1 in antisense orientation prevented prolactin increase in endothelial cell proliferation. Similarly, addition of stannic mesoporphyrin, the inhibitor of HO activity, prevented PRL-mediated increase in endothelial cell proliferation. Our results demonstrated for the first time, that prolactin-mediated angiogenesis and cell proliferation was dependent on HO-1 gene expression.     

Overexpression of heme oxygenase-1 in murine melanoma: increased proliferation and viability of tumor cells, decreased survival of mice

Heme oxygenase-1 (HO-1), a cytoprotective enzyme, can be induced in tumors in response to anti-cancer therapies. We investigated the role of HO-1 in B16(F10), S91, and Sk-mel188 melanoma cells. Overexpression of HO-1 after transduction with adenoviral vectors increased cell proliferation, resistance to oxidative stress generated by H2O2, and angiogenic potential as determined by induction of endothelial cell divisions. Likewise, cells stably transfected with HO-1 cDNA (B16-HO-1) showed higher proliferation, stress resistance, and angiogenic activity than the wild-type line (B16-WT). HO-1 overexpression in tumors significantly shortened survival of mice after subcutaneous injection of cancer cells (38 and 22 days for B16-WT and B16-HO-1, respectively; P=0.017). This also resulted in development of more packed tumors, with more melanoma cells, and reduced inflammatory edemas. Mice injected with B16-HO-1 had lower levels of tumor necrosis factor and higher serum concentrations of its soluble receptor tumor necrosis factor-RI, whereas tumors overexpressing HO-1 displayed augmented vascularization and stronger production of vascular endothelial growth factor. Finally, B16-HO-1 cells injected intravenously formed more metastases in lungs. Thus, HO-1 overexpression increased viability, proliferation, and angiogenic potential of melanoma cells, augmented metastasis, and decreased survival of tumor-bearing mice, suggesting that induction of HO-1 may be detrimental in anti-cancer therapy of melanoma.     

花姜酮通过抑制氧化应激减轻MPP~+诱导的SH-SY5Y细胞损伤

目的:探讨花姜酮(zerumbone,ZER)对1-甲基-4-苯基-吡啶鎓离子(1-methyl-4-phenylpyridinium,MPP~+)诱导的人神经母细胞瘤SH-SY5Y细胞损伤的作用及其可能的分子机制。方法:采用CCK-8法检测MPP~+对SH-SY5Y细胞的毒性作用以及ZER的保护作用,采用流式细胞术检测活性氧簇(reactive oxygen species,ROS)含量和细胞凋亡情况,采用短发夹RNA(short hairpin RNA,shRNA)干扰技术抑制人帕金森病蛋白7(Parkinson disease protein 7,PARK7)基因的表达,采用Western blot法检测PARK7、核因子E2相关因子2(nuclear factor E2-related factor 2,Nrf2)和血红素氧合酶1(heme oxygenase-1,HO-1)的蛋白表达水平。结果:MPP~+可显著抑制SH-SY5Y细胞活力(P0.05),并呈剂量和时间依赖性;ZER可使经600μmol/L MPP~+处理24 h的SH-SY5Y细胞活力升高,PARK7和Nrf2蛋白表达水平显著升高(P0.05),ROS含量和细胞凋亡明显减少(P0.05),抗氧化剂N-乙酰-L-半胱氨酸(N-acetyl-L-cysteine,NAC)具有相似的作用;沉默PARK7基因后,Nrf2和HO-1蛋白的表达水平明显下降(P0.05),ROS含量和细胞凋亡显著增加(P0.05)。结论:ZER可在体外剂量依赖性地减轻MPP~+对SH-SY5Y细胞的毒性作用,这可能与ZER激活PARK7/Nrf2/HO-1通路,继而抑制MPP~+诱导的ROS生成和细胞凋亡有关。     

A protective role for heme oxygenase-1 in INS-1 cells and rat islets that are exposed to high glucose conditions

Heme oxygenase-1 (HO-1) has been described as an inducible protein that is capable of cytoprotection via radical scavenging and the prevention of apoptosis. Chronic exposure to hyperglycemia can lead to cellular dysfunction that may become irreversible over time, and this process has been termed glucose toxicity. Yet little is known about the relation between glucose toxicity and HO-1 in the islets. The purposes of the present study were to determine whether prolonged exposure of pancreatic islets to a supraphysiologic glucose concentration disrupts the intracellular balance between reactive oxygen species (ROS) and HO-1, and so this causes defective insulin secretion; we also wanted to evaluate a protective role for HO-1 in pancreatic islets against high glucose levels. The intracellular peroxide levels of the pancreatic islets (INS-1 cell, rat islet) were increased in the high glucose media (30 mM glucose or 50 mM ribose). The HO-1 expression was induced in the INS-1 cells by the high glucose levels. Both the HO-1 expression and glucose stimulated insulin secretion (GSIS) was decreased simultaneously in the islets by treatment of the HO-1 antisense. The HO-1 was upregulated in the INS-1 cells by hemin, an inducer of HO-1. And, HO-1 upregulation induced by hemin reversed the GSIS in the islets at a high glucose condition. These results suggest HO-1 seems to mediate the protective response of pancreatic islets against the oxidative stress that is due to high glucose conditions.