内、外源性硫化氢在脂多糖所致大鼠急性肺损伤中的作用

目的：探讨内、外源性硫化氢（H2S）在脂多糖（LPS）所致大鼠急性肺损伤（ALI）中的作用并初探其机制。方法：将120只SD大鼠随机分为对照组、IPS组（经气管内滴注LPS复制ALI模型）、NaHS＋LPS组和炔丙基甘氨酸（PPG）＋LPS组。给药后4h或8h处死动物,测定肺系数;光镜观察肺组织形态学改变;化学法检测血浆H2S、NO和CO含量、肺组织丙二醛（MDA）含量、胱硫醚-γ-裂解酶（CSE）、诱导型一氧化氮合酶（iNOS）和血红素加氧酶（HO）活性以及支气管肺泡灌洗液（BALF）中中性粒细胞（PMN）数目和蛋白含量的变化;用免疫组织化学法检测肺组织iNOS、HO-1蛋白表达。再将血浆H2S含量与上述指标进行相关性分析。结果：气管内滴注LPS可引起肺组织明显的形态学改变;肺系数和肺组织MDA含量增加;BALF中PMN数目和蛋白含量增加;血浆H2S含量和肺组织CSE活性下降;肺组织iNOS活性、HO活性和iNOS蛋白表达、HO-1蛋白表达增强,血浆NO含量、CO含量增加。预先给予NaHS可显著减轻LPS所致上述指标的改变;而预先给予PIG可加重LPS所致肺损伤,使BALF中PMN数目和蛋白含量、血浆NO含量、肺组织iNOS活性和iNOS蛋白表达进一步增加,但对血浆CO含量、肺组织HO活性和HO-1蛋白表达无明显影响。HS含量与CSE活性、血浆CO含量、肺组织HO-1活性呈正相关（r值=0．945—0．987,P均〈0．01）;与其他指标呈负相关（r值=-0．994～-0．943,P均〈0．01）。结论：H2S／CSE体系的下调在LPS所致大鼠Ⅲ的发病学中有一定作用,内、外源性H2S具有抗LPS所致Au的作用,该作用可能与其抗氧化效应、减轻PMN所致肺过度的炎症反应以及下调NO／iNOS体系、上调CO／HO—1体系有一定关系。     

一氧化碳在低氧性肺血管收缩反应中的作用

目的和方法 :观察外源性一氧化碳 (CO)对大鼠离体肺动脉环低氧性收缩反应 (HPV)的影响 ,并通过观察血红素氧化酶抑制剂ZnPPIX对HPV的影响 ,探讨内源性一氧化碳在HPV中的作用及机制。结果 :低氧可使苯肾上腺素 (PE)预收缩的肺动脉环出现明显的收缩反应 ,肺动脉cGMP含量下降 ;用ZnPPIX孵育后 ,低氧后的肺动脉cGMP含量增加 ,低氧性肺血管收缩反应 (HPV)受抑 ;外源性CO可明显增加肺动脉cGMP含量 ,HPV明显受抑。结论 :外源性CO及ZnPPIX可增加低氧后的肺动脉cGMP含量 ,抑制HPV ,内源性CO减少导致cGMP含量下降可能是HPV的原因之一     

内源性CO在缺氧性肺动脉高压大鼠肺血重构中的作用

目的和方法：应用逆转录聚合酶链式反应（RT－PCR）、双波长分光光度法,右心导管及维多利亚蓝染色方法,动态观察慢性缺氧不同时间点大鼠肺吕诱导型血红素氧合酶（HO－1）基因表达,内源性CO生成,肺动脉压力及构型的变化,探讨内源性CO大鼠缺氧性肺动脉高压肺血管重构中的作用。结果：（1）正常大鼠肺组织可表达少量HO－1mRNA,缺氧5、10、15d大鼠肺组织HO－1mRNA含量分别增加2．3、3．6、4．0倍（P＜0．01）,动脉血中COHb分别较正常大鼠增加1．2、2．6和2．9倍（P＜0．1或P＜0．05）。同时RVSP升高。光镜下可见IAPA血管壁增厚,管腔变窄。（2）Hemin可使缺氧大鼠肺组织HO－1mRNA和动脉血中COHb保持在高水平（分别高达正常对照组的5．2和3．7倍,P＜0．01或P＜0．05）,能部分地抑制缺氧时大鼠RVSP的升高,减轻IAPA的病理改变。结论：在慢性缺氧性肺动脉高压大鼠肺组织中HO－1基因的表达增加,内源性CO生成增多。Hemin促进HO－1基因表达和内源性CO手成,可抑制肺动脉压升高,阻抑制血管重构,对缺陷氧性肺动脉高压的形成有一定的防治作用。     

葛根素对抗高糖诱导的血管低反应性的作用及其机制

目的：研究葛根素是否可对抗高糖引起的血管低反应性,并探讨其作用机制。方法：采用血管环离体灌流装置,观察SD大鼠胸主动脉环的收缩反应;测定主动脉胆红素生成量反映血红素加氧酶-1（heme oxygenase-1,HO-1）的活性。结果：①与空白对照组（含11 mmol/L葡萄糖）相比,经44 mmol/L葡萄糖（高糖）孵育血管4 h后,主动脉环对苯肾上腺素（PE）引起的血管收缩反应下降;且该作用通过内皮依赖性途径实现。②葛根素（10-10~10-8mol/L）与高糖联合孵育,可剂量依赖性地改善高糖诱导的血管PE收缩反应的下降。③葛根素孵育血管后可引起血管HO-1活性增高;用ZnPP抑制HO-1的活性后,葛根素抗高糖损伤的作用被取消。结论：葛根素具有对抗高糖引起的血管收缩功能下降的作用,其机制可能是通过诱导HO-1活性增加实现的。     

人体血红素加氧酶-1的研究进展

血红素加氧酶（heme oxygenase,HO)是哺乳动物中血红素代谢的限速酶,HO－1是HO同功酶之一,主要分布在肝、脾、肺等多种脏器,具有调节和保护功能。作者拟从人体HO－1蛋白的晶体结构、HO－1的功能和HO－1表达的诱导因素,以及HO－1基因的表达与调控等研究进展做一综述。     

转染人血红素加氧酶-1基因对大鼠血管平滑肌细胞抗氧化损伤的作用

本实验构建含人血红素加氧酶-1（hHO-1)基因的逆转录病毒载体XM-6/hHO-1,将其导入离体培养的大鼠血管平滑肌细胞（vascular smooth muscle cells,VSMC),观察外源性hHO-1基因在VSMC内的表达及其抗活性氧损伤作用,结果表明：（1）hHO-1基因可在靶细胞中明显表达,转染VSMC的HO-1蛋白表达和HO酶活性分别比非转染细胞高1.8倍和2.0倍;（2）转染hHO-1的VSMC可对抗大剂量H2O2对细胞的损伤作用,表现为细胞存活率增加和乳酸脱氢酶（LDH）漏出减少,上述保护作用可被HO特异性抑制剂锌原卟啉IX（Zinc-proto-porphyrinIX,ZnPP-IX)所阻断,研究结果提示,外源性HO-1的过量表达可增加VSMC对抗氧化损伤的能力。     

17β-雌二醇对去卵巢胰岛素抵抗大鼠主动脉舒缩功能损伤的保护作用

为观察17β-雌二醇（17beta-estradiol,17β-E2）对去卵巢胰岛素抵抗（insulin resistance,IR）大鼠主动脉结构和舒缩功能的影响及其可能机制,成年雌性Sprague-Dawley大鼠卵巢切除后,高果糖喂养8周诱导IR,同时给予生理剂量的17β-E2（30μg／kg）,每天皮下注射一次,并检测IR相关指标。大鼠胸主动脉石蜡切片,HE染色,图像分析系统测定其结构。采用血管环灌流法,观察各组大鼠胸主动脉环对新福林（L-phenylephrine,PE）的收缩反应和对ACh、硝普钠（sodium nitroprusside,SNP）的舒张反应以及一氧化氮合酶（nitric oxide synthase,NOS）抑制剂N-硝基-L-精氨酸甲脂（N-nitrl-L-arginine methylester,L-NAME）对卵巢切除＋果糖喂养＋17β-E2组大鼠胸主动脉ACh的舒张反应的影响;检测各组大鼠一氧化氮（nitric oxide,NO）含量。结果显示：（1）17β-E2能防止高果糖诱导的去卵巢IR人鼠收缩压升高、高胰岛素血症和胰岛素敏感性下降;（2）各组火鼠胸主动脉的结构无显著性差异;（3）卵巢切除＋果糖喂养组大鼠与卵巢切除组或果糖喂养组相比,血清NO显著降低,胸主动脉对PE的收缩反应显著增强,对ACh的舒张反应显著降低,17β-E2能逆转上述改变,L-NAME可部分阻断17β-E2的这种作用;（4）各组大鼠胸主动脉对SNP的舒张反应和去内皮后对PE的收缩反应均无显著差异。以上结果表明,17β-E2能抑制高果糖诱导的去卵巢IR大鼠血管舒缩功能的紊乱,其机制一方面可能是部分通过血管内皮细胞NOS途径促进NO的释放,保护内皮细胞;另一方面可能是通过降低血压,血清胰岛素水平,改善IR所致。     

内毒素引起的乳鼠心肌细胞血红素加氧酶—1基因的表达

为了探讨在内毒素作用下的乳鼠心肌细胞（neonatal rat cardiomyocytes,NRCMs）血红素加氧酶－1（heme oxygenase-1,HO-1）基因的表达及其在细胞损伤中的作用,分别用10、30及50μg/ml的脂多糖（lipopolysaccharide,LPS）,10μg/ml LPS 10μmol/ml锌原卟啉Ⅸ（Zn-protoporphyrin-Ⅸ,ZnPPⅨ）和单纯10μmol/ml ZnPPⅨ与培养的NRCMs共同孵育6h,以及10μg/ml LPS与NRCMs共同孵育9h和18h。分别观察细胞HO－1 mRNA表达、MDA含量、LDH释放量与台盼蓝摄取率的变化。结果显示,同样与细胞孵育6h,LPS10μg/ml时HO－1 mRNA表达比对照组增加81.2％,30μg/ml时表达量增加126.3％,50μg/ml时表达量增加92.8％;LPS为10μg/ml时,孵育9h后HO－1 mRNA的表达量比对照组增加93.6％,孵育18h后一增加105.8％。LPS30、50μg/ml,10μg/ml LPS＋10μmol/ml ZnPPⅨ与细胞孵育6h及LPS 10μg/ml孵育18h后,细胞MDA含量、LDH释放量与台盼蓝摄取率明显增加（P＜0.01）;单纯10μg/ml LPS与单纯10μmol/ml ZnPPⅨ孵育6h后,上述指标均无明显升高。结果表明,LPS可诱导NRCMs HO－1 mRNA的表达,且在较低LPS剂量范围内具有时间依赖性和浓度依赖性;NRCMs HO-1 mRNA的表达可减低LPS引起的细胞损伤,这可能是细胞产生的一种自身保护性反应。     

知母宁对慢性低O2高CO2大鼠肺动脉高压的影响及其机制研究

目的：研究知母宁对慢性低氧高二氧化碳性肺动脉高压的抑制作用及其作用机制。方法：将SD大鼠分为正常对照组,4周低O2高CO2组,4周低O2高CO2^ 知母宁组（知母宁组）。用透射电镜、图像分析、免疫组化、组织原位杂交技术等方法,观察各组大鼠肺动脉平均压（mPAP)、颈动脉平均压（mCAP)、肺细小动脉显微和超微结构、血CO浓度、血红素氧合酶－1（HO－1）及其基因表达的变化。结果：①低O2高CO2组mPAP比正常组显著增高,知母宁组mPAP比低O2高CO2组显著降低;3组间mCAP比较差异无显著性。②全血CO浓度低O2高CO2组比正常组显著增高,知母宁组比低O2高CO2组增高。③光镜下低O2高CO2组与正常组相比,肺细小动脉管壁面积／管总面积、肺细小动脉中膜平滑肌细胞核密度、肺细小动脉中膜厚度均显著升高,知母宁组与低O2高CO2组相比以上指标显著降低。④电镜下知母宁组肺细小动脉中膜平滑肌细胞增生、面积增大、染色质增多、外膜胶原纤维增生均较低O2高CO2组明显减轻。⑤免疫组化、原位杂交发现低O2高CO2组大鼠肺细小动脉HO－1及mRNA与正常组比较均明显增加,知母宁组大鼠肺细小动脉HO－1及mRNA表达均较低O2高CO2组为高。结论：知母宁有抑制慢性低氧高二氧化碳性肺动脉高压和肺血管结构重建的作用,上调HO－1及其基因表达、使CO合成增多可能为其重要作用机制。     

血红素加氧酶-1在对抗过氧化氢引起的血管低反应性中的作用

目的：研究HO-1的诱导剂是否可对抗H2O2引起的血管低反应性,并探讨其作用机制。方法：采用血管环灌流装置,观察胸主动脉环的收缩效应。结果：①SD大鼠腹腔注射高铁血红素后,主动脉HO-1活性和血中CO含量增高;同时,H2O2引起的血管收缩功能下降的现象明显改善。②KATP通道阻断剂优降糖,而非GC抑制亚甲蓝,可取消高铁血红素的抗H2O2损伤的作用。③Hemin＋H2O2组与单纯H2O2组的钙收缩曲线无明显差异。④无钙液中,高铁血红素可抑制H2O2引起的咖啡因和PE诱导的收缩幅度的下降。结论：诱导主动脉HO-1活性增加,可对抗氧化应激引起的血管收缩反应的低下,其机制可能是通过激活KATP通道,影响细胞内贮存钙的释放起作用。而与GC信号转导通路无关。     

Changes of heme oxygenase-carbon monoxide system in vascular calcification in rats

The aim of the present study was to investigate the change in heme oxygenase (HO)-carbon monoxide (CO)-cyclic guanosine monophosphate (cGMP) pathway in vascular calcification. Vascular calcification model was established in rats by using vitamin D(3) and nicotine. Vascular calcium content, alkaline phosphatase (ALP) activity, HO activity, HbCO formation and content of cGMP in vessels were measured. Immunochemistry (IH) for HO 1 expression and in situ hybridization (ISH) for HO 1 mRNA were observed. Compared to those of control rats, the aortic calcium content and vascular ALP activity in rats of the calcified group (VDN group) were obviously increased, but HO 1 activity, CO concentration and cGMP content in vessels of rats in VDN group were markedly decreased. Expressions of HO-1 protein and mRNA were significantly decreased compared to control rats. Vascular calcification might induce a down regulation in vascular HO-CO-cGMP pathway.     

Renal vasodilatory influence of endogenous carbon monoxide in chronically hypoxic rats

Chronic hypoxia (CH) attenuates systemic vasoconstriction to a variety of agonists in conscious rats. Recent evidence suggests that similarly diminished responses to vasoconstrictors in aortic rings from CH rats may be due to increased endothelial heme oxygenase (HO) activity and enhanced production of the vasodilator carbon monoxide (CO). Thus we hypothesized that a hypoxia-induced increase in HO activity is responsible for decreased vasoconstrictor responsiveness observed in conscious CH rats. CH (4 wk at 0.5 atm) and control rats were renal denervated and instrumented for the measurement of renal blood flow (RBF) and blood pressure. First, renal vasoconstrictor responses to graded intravenous infusion of phenylephrine (PE) were assessed in conscious rats. CH rats demonstrated significantly diminished renal vasoconstrictor responses to PE compared with control responses that persisted even with acute restoration of normoxia. In additional experiments, CH rats exhibited increased renal vascular resistance and decreased RBF in response to the HO inhibitor zinc protoporphyrin IX (11 micromol/kg iv), whereas renal hemodynamics were unaffected by the inhibitor in control animals. Furthermore, we demonstrated greater HO enzyme activity in renal tissue from CH rats compared with controls. These data suggest that enhanced HO activity contributes a tonic vasodilatory influence in the renal vasculature of CH rats that may be responsible for the diminished sensitivity to vasoconstrictor agonists observed under these conditions.     

Hyperpolarization contributes to vascular hyporeactivity in rats with lipopolysaccharide-induced endotoxic shock

We have examined the role of membrane hyperpolarization in mediating vascular hyporeactivity induced by bacterial lipopolysaccharide (LPS) in endothelial-denuded strips of rat thoracic aorta ex vivo. The injection of rats with LPS caused a significant fall of blood pressure and a severe vascular hyporeactivity to norepinephrine. The membrane potential recording showed that endotoxemia caused a hyperpolarization when compared to the control. This hyperpolarization was fully restored by methylene blue (MB; 10 microM) and partially reversed by Nomega-nitro-L-arginine methyl ester (L-NAME; 0.3 mM), 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ; 1 microM), tetraethylammonium (TEA; 10 mM), charybdotoxin (CTX; 0.1 microM), or glibenclamide (GB; 10 microM), however, this hyperpolarization was not significantly affected by apamin (0.1 microM), 4-aminopyridine (4-AP; 1 mM), or Ba2+ (50 microM). In addition, the basal tension of the tissues obtained from endotoxemic rats was enhanced by the following order: MB > or = ODQ > TEA > or = L-NAME > or = CTX > GB; whereas apamin, 4-AP or Ba2+ had no significant effects on these tissues. In contrast, none of these inhibitors had significant effects on the membrane potential or the basal tension in control tissues. Our electrophysiological results further confirmed previous studies showing that in addition to nitric oxide, the large conductance Ca2+-activated K+-channels and ATP-sensitive K+-channels are, most likely, responsible for endotoxin-mediated hyporeactivity to vasoconstrictor agents in vascular smooth muscle.     

褪黑素改善内毒素血症大鼠血管反应性

观察褪黑素(melatonin,MT)对脂多糖(lipopolysaccharide,LPS)诱导的体循环和肺循环血管反应性失调的影响,并探讨可能的作用机制。实验分为溶剂对照组、LPS组、LPS+MT组和MT组。制备离体胸主动脉环和肺动脉环,应用血管张力检测技术检测各组血管环对苯肾上腺素(phenylephrine,PE)和乙酰胆碱(acetylcholine,ACh)的反应性并绘制累积剂量反应曲线;制备各组血管组织匀浆,测定丙二醛(malondialhyde,MDA)和超氧化物歧化酶(superoxidedismutes,SOD)含量变化。结果显示:与对照组相比,LPS6h后胸主动脉对PE的收缩反应减弱(P<0.01),对PE(1×10–8~1×10–5mol/L)累积剂量反应曲线下移;而肺动脉对ACh的舒张反应显著下降(P<0.01),对ACh(1×10–8~1×10–5mol/L)累积剂量反应曲线下移。加用MT可显著改善LPS诱导的胸主动脉对缩血管剂PE的低反应性,同时可逆转LPS对肺动脉舒张反应的抑制,LPS+MT组胸主动脉对PE的累积剂量反应曲线和肺动脉对ACh的累积剂量反应曲线位于对照组和LPS组之间;MT还可对抗LPS导致的脂质过氧化,使MDA含量减少,提高抗氧化酶SOD的活性。上述结果提示,MT可改善内毒素血症大鼠的血管反应性失调,抗氧化途径可能是其发挥保护作用的机制之一。     

Heme oxygenase modulates selectin expression in different regional vascular beds

Heme oxygenase (HO) catalyzes the degradation of heme to biliverdin, iron, and CO. The inducible isoform (HO-1) has been implicated as a modulator of the inflammatory response. HO-1 activity can be induced by hemin and inhibited with zinc protoporphyrin IX (ZnPP). Using these reagents, we assessed the possibility that HO-1 modulates the inflammatory response by altering the expression of endothelial cell adhesion molecules. Endotoxin (lipopolysaccharide, LPS)-induced expression of P- and E-selectin expression was quantified in different vascular beds of the rat using the dual radiolabeled monoclonal antibody technique. Pretreatment with hemin attenuated, whereas ZnPP treatment exacerbated, the increased selectin expression normally elicited by LPS. Biliverdin, at an equimolar dosage, was as effective as hemin in attenuating LPS-induced selectin expression in the lung, kidneys, liver, and intestines. These findings indicate that the anti-inflammatory properties of HO-1 may be related to an inhibitory action of P- and E-selectin expression in the vasculature. Biliverdin (or its metabolite, bilirubin), rather than CO, may account for this action of HO-1 on endothelial cell adhesion molecule expression.     

Endogenous carbon monoxide is an endothelial-derived vasodilator factor in the mesenteric circulation

Chronic hypoxia (CH) is associated with both blunted agonist-induced and myogenic vascular reactivity, possibly due to an enhanced production of heme oxygenase (HO)-derived carbon monoxide (CO). However, the cellular location of the HO responsible for these effects has not been clearly established. Therefore, we examined the response to administration of the substrate for HO, heme-l-lysinate (HLL), in endothelium-intact and endothelium-denuded small mesenteric arteries from CH male Sprague-Dawley rats. Mesenteric arteries were isolated and mounted on glass cannulas, pressurized to 60 mmHg, and superfused with physiological saline solution. All experiments were performed in the presence of 100 microM N(omega)-nitro-l-arginine. The vasodilator response to HLL or exogenous CO was examined. HLL experiments were performed in the presence and absence of the HO inhibitor zinc protoporphyrin IX (ZnPPIX). HLL administration resulted in a dose-dependent vasodilator response that was abolished in the presence of ZnPPIX or by endothelial removal. Exogenous CO produced a vasodilator response that was independent of an intact endothelium. Cellular localization of HO was verified through immunohistochemistry in sections of the gut and aorta from CH and control animals. Staining for HO-1, HO-2, and endothelial nitric oxide synthase was confined to the endothelium. Thus we conclude that CO is a product of HO located within the endothelium.     

Inhibition of endogenous CO by ZnPP protects against stress-induced gastric lesion in adult male albino rats

Carbon monoxide (CO) has been found to be produced in every living cell in a biochemical reaction catalyzed by heme-oxygenase (HO) enzyme which degrades heme into biliverdin, CO, and iron. Endogenous CO is not a waste product, but acts as a chemical messenger mediating and modulating many intracellular biochemical reactions that regulate physiological functions. This study was designed to investigate the effect of inhibition of endogenous CO production by zinc protoporphyrin (ZnPP), an HO inhibitor, on the gastric secretion and ulceration induced by cold-restraint stress (CRS) in adult male albino rats. Rats were pylorically ligated and divided randomly into the following groups (six rats each): control, ZnPP treated (50 μmol/kg/day, s.c. for 10 days), CRS, and stressed ZnPP treated groups. Blood samples were collected from the retro-orbital sinus of anesthetized rats for determination of CO concentration. We found that ZnPP pretreatment significantly decreased HO-1 level, CO level, and volume of gastric juice as compared to the control non-stressed rats. In the present study, ZnPP pretreatment proved to be protective against development of ulcerative lesions in CRS model as evidenced by reduction of the ulcer index, and this could be mediated through reduction of free and total acidity of gastric secretion and decreased lipid peroxidation but with significantly decreased gastric protective nitric oxide and prostaglandin E(2) levels. In conclusion and according to our results, the protective effect of ZnPP on CRS-induced gastric ulcers despite of inhibition of endogenous CO could be attributed to the presence of zinc which is known to have a protective anti-ulcer effect.     

内源性一氧化碳减轻大鼠双侧后肢缺血再灌注所致的肺损伤

通过观察血红素氧化酶（HO）阻断剂－－锌原卟啉（ZnPP)对肺组织、肺泡间质多形核白细胞数目肺组织丙二醛含量和湿重干重之比的影响,并对肺组织HO活性和血内碳氧血红蛋白水平（COHb）进行检测,以探讨内源性HO／一氧化碳（CO）在肢体缺血再灌注（I／R）所致肺损伤中的作用。结果发现,大鼠双侧后肢I／R可导致急性肺损伤,同时使肺组织中HO活性和血内COHb水平显著升高;应用ZnPP预处理可使HO活性和COHb水平显著降低,但肺损伤却进一步加重。上述实验结果表明,肢体I／R致肺损伤时,肺组织中HO活性和内源性CO生成增多或减轻大鼠肢体I／R所致的肺损伤。     

Zinc Protoporphyrin Regulates Cyclin D1 Expression Independent of Heme Oxygenase Inhibition

Zinc protoporphyrin IX (ZnPP), an endogenous heme analogue that inhibits heme oxygenase (HO) activity, represses tumor growth. It can also translocate into the nucleus and up-regulate heme oxygenase 1 (HMOX1) gene expression. Here, we demonstrate that tumor cell proliferation was inhibited by ZnPP, whereas tin protoporphyrin (SnPP), another equally potent HO-1 inhibitor, had no effect. Microarray analysis on 128 tumorigenesis related genes showed that ZnPP suppressed genes involved in cell proliferation and angiogenesis. Among these genes, CYCLIN D1 (CCND1) was specifically inhibited as were its mRNA and protein levels. Additionally, ZnPP inhibited CCND1 promoter activity through an Sp1 and Egr1 overlapping binding site (S/E). We confirmed that ZnPP modulated the S/E site, at least partially by associating with Sp1 and Egr1 proteins rather than direct binding to DNA targets. Furthermore, administration of ZnPP significantly inhibited cyclin D1 expression and progression of a B-cell leukemia/lymphoma 1 tumor in mice by preferentially targeting tumor cells. These observations show HO independent effects of ZnPP on cyclin D1 expression and tumorigenesis.