姜黄素诱导血红素氧合酶-1表达抗大鼠主动脉平滑肌细胞增殖作用的研究

目的:探讨姜黄素诱导大鼠主动脉平滑肌细胞(RASMC)血红素氧合酶-1(HO-1)表达效果及其抗细胞增殖的作用.方法:RASMC株经复苏、传代培养后分组实验,分别加用不同浓度的姜黄素或姜黄素 ZnPPⅨ孵育,用Western blot法检测HO-1表达效果;用四唑氮法及3H-Td掺入法检测RASMC增殖情况;同时还观察了姜黄素的细胞毒性作用.结果:姜黄素能呈剂量依赖性诱导RASMC高表达HO-1蛋白并显著抑制血清刺激的RASMC增殖(P＜0.05或P＜0.01),加入ZnPPⅨ共同孵育能使姜黄素诱导HO-1表达明显降低并消除其对RASMC增殖的抑制作用(均P＜0.01),实验浓度的姜黄素无明显细胞毒性作用(P＞0.05).结论:姜黄素能显著诱导RASMC表达HO-1蛋白并以此有效抑制RASMC增殖.     

血红素加氧酶-1与糖尿病心肌病变

糖尿病心肌病变是一种独立于其它心血管并发症如高血压、冠心病之外的心脏病,氧化应激主导了糖尿病心肌病变的发病过程.血红素加氧酶(HO)-1,是体内参与亚铁血红素代谢的一种具有多种功能的微粒体氧化酶,在氧化应激过程中起重要的细胞保护作用,它的诱导已成为氧化应激的一种普遍标志.进一步探讨在糖尿病心肌病变中氧化应激与HO-1的关系对于预防和治疗糖尿病心肌病变具有重要价值.     

低氧对间充质干细胞中血红素加氧酶1表达的影响

目的:探讨缺氧对人骨髓间充质干细胞血红素加氧酶-1(HO-1)表达的影响。方法:采用percoll密度离心法自健康人骨髓中分离MSCs并进行纯化和扩增,通过流式细胞术检测其表面标志进行鉴定;用CoCl2建立化学低氧模型,通过实时荧光定量PCR检测低氧下HO-1mRNA表达的量-效和时-效关系。结果:流式细胞术检测MSCs细胞表面抗原结果显示高表达CD13、CD44、CD71,而CD34、CD45表达阴性。CoCl2模拟的化学性低氧促进了人骨髓间充质干细胞中HO-1的表达,并且这种表达呈现出较好的时间、剂量依赖性。结论:CoCl2诱导慢性缺氧可促进人骨髓间充质干细胞的HO-1基因表达。     

血红素加氧酶-1在肾脏疾病中的研究进展

血红素加氧酶（HO）是血红素降解的起始酶和限速酶,其主要功能是代谢血红素成为一氧化碳、胆绿素及铁离子。HO—1是HO的一种同工酶,为诱导型HO。HO-1及血红素代谢产物具有抗炎、抗氧化、抗凋亡、扩张血管等生理作用。HO-1作为一种重要的应激蛋白,在急性肾脏损伤、肾小球肾炎、肾间质纤维化等肾脏疾病的病理生理发展过程中发挥着重要的保护作用,有效地调控其表达有可能成为预防和治疗相关疾病的新策略。     

血红素加氧酶-1在非酒精性脂肪性肝炎进展中的作用

目的 探讨血红素加氧酶-1(HO-1)在非洒精性脂肪性肝炎进展中的作用及其机制.方法 选用健康雄性C57BL/6J小鼠,采用胆碱-蛋氨酸缺乏饮食(MCD)4周建立小鼠非酒精性脂肪性肝炎模型,以胆碱-蛋氨酸充足饮食设立对照组,并以MCD加HO-1激动剂血晶素或抑制剂锌原卟啉进行干预实验.小鼠血清ALT和AST采用全自动生化仪酶法测定.HE染色观察肝脂肪变、炎症活动及纤维化程度;逆转录聚合酶链反应和Western blot检测HO-1、肿瘤坏死因子(TNF)α和白细胞介素(IL)-6 mRNA及其蛋白的表达.结果 MCD喂养小鼠血清ALT及AST明显异常,出现中～重度肝细胞脂肪变性,伴有点状和灶状肝细胞坏死、炎性细胞浸润、轻度汇管区纤维组织增生及窦周纤维化;HO-1、TNF α和IL-6 mRNA及其蛋白的表达较对照组显著增强,相对表达量分别为1.13±0.11、1.74±0.05,0.20±0.01、1.92±0.10,0.58±0.02、2.06±0.05对比0.43±0.02、0.75±0.05,0.08±0.00、0.59±0.02,0.22±0.01、0.91±0.02(P＜0.01);应用血晶素小鼠随肝脏HO-1 mRNA及其蛋白表达的上调及TNF α和IL-6 mRNA及其蛋白表达的下调(P＜0.01),肝脂肪变及炎症活动度均显著减轻;而应用锌原卟啉小鼠,肝脏HO-1 mRNA及蛋白表达明显受抑制,TNF α和IL-6 mRNA及蛋白表达则明显增强(P＜0.01),肝脂肪变及炎症亦随之显著加重.结论 抗氧化基因HO-1靶向性激活可阻止非酒精性脂肪性肝炎的发生及进展.     

血红素加氧酶-1在糖尿病大鼠心肺、血管组织的表达

目的观察和比较糖尿病大鼠血管、心脏和肺脏组织中血红素加氧酶-1(HO-1)表达水平。方法应用RT-PCR和免疫组织化学技术检测链脲佐菌素诱导的糖尿病大鼠心脏、肺脏和胸主动脉HO-1mRNA和蛋白的表达。结果与对照组相比,糖尿病大鼠心脏和肺脏组织的HO-1表达增强,而胸主动脉HO-1表达无差异。结论与心脏、肺脏相比较,血管组织不能在糖尿病状态下有效表达HO-1。     

早产儿血清血红素加氧酶-1与支气管肺发育不良的关系

目的 探讨血清血红素加氧酶-1(HO-1)在支气管肺发育不良(BPD)早产儿早期诊断中的价值.方法 前瞻性选取胎龄≤32周的早产儿,于出生时、生后2周时采集静脉血检测血清HO-1水平.根据是否发生支气管肺发育不良将患儿分为BPD组(包括轻度组、中/重度组)和非BPD组,从非BPD组中筛选出与BPD组一般情况差异无统计学...     

血红素加氧酶-1对糖尿病大鼠胸主动脉功能的影响

目的探讨血红素加氧酶-1（HO-1）对糖尿病（DM）大鼠大血管功能的影响。方法SD大鼠分为4组：对照组、DM组、hemin（HO-1诱导剂）＋DM组、锌原卟啉（ZnPP,HO-1抑制剂）＋DM组。RT-PCR法检测血管组织HO-1表达水平 应用离体血管张力检测技术观察胸主动脉血管的反应性 扫描电镜下观察血管内皮超微结构。结果hemin＋DM组HO-1mRNA水平是对照组的2.01倍。与DM组相比,hemin＋DM组血管环对乙酰胆碱（Ach）舒张百分率增高,对苯肾上腺素（PE）的收缩反应减弱,内皮细胞形态有所改善 而ZnPP＋DM组大鼠对Ach的舒张反应继续下降,内皮细胞破坏严重。结论提高HO-1的表达水平有益于改善DM大鼠大血管反应性失调。     

糖尿病足溃疡皮肤血红素加氧酶1表达的研究

目的 探讨血红素加氧酶1(HO-1)在糖尿病足溃疡皮肤中的表达.方法 RT-PCR检测非糖尿病的足溃疡(NDFU)组、糖尿病足溃疡(DFU)组、正常对照(NC)组皮肤HO-1mRNA的表达;免疫组化检测各组HO-1蛋白表达.结果 NDFU组、DFU组、NC组HO-1mRNA表达量分别为:1.880±0.033、1.797±0.024、1.399±0.039,三组间差异有统计学意义;NDFU组和DFU组比较,差异有统计学意义(P均<0.01).HO-1蛋白在NC组表达于表皮;在NDFU组和DFU组表皮和真皮炎症部位均有HO-1阳性细胞,且前者阳性细胞数量多于后者.结论 糖尿病足溃疡较非糖尿病的足溃疡皮肤组织HO-1表达降低.     

血红素加氧酶-1在肝脏缺血再灌注损伤研究中新进展

肝脏缺血再灌注损伤(hepatic ischemia reperfusion injury,HIRI)的分子机制迄今尚未完全清楚。血红素加氧酶-1(heme oxygenase-1,HO-1)是体内血红素降解的起始酶和限速酶,在体内分解血红素生成一氧化碳(CO)、胆绿素和自由铁。HO-1系统具有抗炎、抗氧化、抗凋亡和促进细胞存活、循环稳定和免疫调节的作用,在HIRI中发挥着至关重要的作用。通过药物或基因工程的方法诱导产生HO-1,能减轻HIRI,目前这一方向已经成为该领域的研究热点。     

Prolactin regulates mammary epithelial cell proliferation via autocrine/paracrine mechanism

Prolactin (PRL) is essential for a number of developmental events in the mammary gland. Work with PRL and PRL receptor knockout mice has shown that PRL indirectly regulates ductal side branching during puberty and directly controls lobuloalveolar development and lactogenesis during pregnancy. Anterior pituitary or placental PRL is thought to be responsible for these functions via an endocrine mechanism; however, PRL is also produced in a number of extrapituitary sites including the mammary gland. The physiologic relevance of mammary PRL remains unknown. In this study we utilized mammary recombination in Rag1^−/− hosts, to determine whether mammary PRL plays a role in the regulation of mammary gland development. Mammary glands formed with the PRL gene deleted from either the epithelium, stroma, or both displayed normal development, on the basis of whole mount and hematoxylin and eosin histology, during puberty and lactation. At the end of pregnancy, a 2.8-fold decrease in bromodeoxyuridine incorporation was observed in the epithelial cells of mammary glands formed using PRL knockout epithelium compared with those formed using wildtype epithelium. No balancing alteration in the rates of apoptosis was detected. Thus, mammary-derived PRL influences mammary epithelial cell proliferation via an autocrine/paracrine mechanism, establishing a physiologic function for mammary PRL during mammopoiesis.     

Pterostilbene Decreases Cardiac Oxidative Stress and Inflammation via Activation of AMPK/Nrf2/HO-1 Pathway in Fructose-Fed Diabetic Rats

Purpose

Oxidative stress has a pivotal role in the pathogenesis of diabetes-associated cardiovascular problems, which has remained a primary cause of the increased morbidity and mortality in diabetic patients. It is of paramount importance to prevent the diabetes-associated cardiac complications by reducing oxidative stress with the help of nutritional or pharmacological agents. Pterostilbene (PT), the primary antioxidant in blueberries, has recently gained attention for its promising health benefits in metabolic and cardiac diseases. However, the mechanism whereby PT reduces diabetic cardiac complications is currently unknown.

Methods

Sprague-Dawley rats were fed with 65% fructose diet with or without PT (20 mg kg^?1 day^?1) for 8 weeks. Heart rate and blood pressure were measured by tail-cuff apparatus. Real-time PCR and western blot experiments were executed to quantify the expression levels of mRNA and protein, respectively.

Results

Fructose-fed rats demonstrated cardiac hypertrophy, hypertension, enhanced myocardial oxidative stress, inflammation and increased NF-κB expression. Administration of PT significantly decreased cardiac hypertrophy, hypertension, oxidative stress, inflammation, NF-κB expression and NLRP3 inflammasome. We demonstrated that PT improved mitochondrial biogenesis as evidenced by increased protein expression of PGC-1α, complex III and complex V in fructose-fed diabetic rats. Further, PT increased protein expressions of AMPK, Nrf2, HO-1 in cardiac tissues, which may account for the prevention of cardiac oxidative stress and inflammation in fructose-fed rats.

Conclusions

Collectively, PT reduced cardiac oxidative stress and inflammation in diabetic rats through stimulation of AMPK/Nrf2/HO-1 signalling.

     

Hydrocortisone promotes survival and proliferation of granulocyte-macrophage progenitors via monocytes/macrophages

We have determined the mechanism by which hydrocortisone (Hc) promotes the survival and proliferation of normal human granulocyte-macrophage progenitors (granulocyte-macrophage colony-forming units, CFU-GM). Peripheral blood mononuclear cells were cultured in a liquid system with 0-10.0 microM Hc over 3 weeks. At 7-day intervals 50% of the culture media along with the cells (suspension cells) present in the media were removed and replaced with fresh media. No CFU-GM or very small numbers of CFU-GM were contained in the suspension cells of the 14- and 21-day-old liquid cultures without Hc; CFU-GM were present and increased with increasing concentrations of Hc. The CFU-GM content in suspension cells of 14- and 21-day-old liquid cultures with 1.0 microM Hc was at least threefold higher compared to liquid cultures without Hc. In a double-layer CFU-GM agar culture system, the suspension cells from liquid cultures with 1.0 microM Hc, but not from liquid cultures without Hc, supported CFU-GM proliferation from normal human bone marrow cells. The CFU-GM proliferation-inducing ability was confined to the monocytes/macrophages (Mo). CFU-GM colony inhibitory and stimulatory activities were detected in cell-free media recovered from liquid cultures without Hc, but only colony stimulatory activity was detected in the media from cultures with 1.0 microM Hc. These results indicate that greater than or equal to physiological concentrations of Hc (0.1-1.0 microM) are required for the persistence and proliferation of CFU-GM, and the effect of Hc is mediated through the Mo, probably by inhibiting the production of one or more of the CFU-GM colony inhibitory molecules.     

CDl63／HO-1通路激活在稳定动脉粥样硬化斑块中的作用

动脉粥样硬化（atherosclerosis，AS）目前被广泛认为是一种慢性炎症性疾病，炎症反应与斑块的不稳定性密切相关。CD163作为血红蛋白特异清道夫受体，近年被研究发现可以调节血红素氧合酶-1（hemeoxygenase-1，HO-1）的表达，而后者已经大量实验证明具有抗炎、抗氧化作用。因此有学者提出，CD163／HO-1通路作为一个新的抗炎途径，有望在稳定动脉粥样硬化不稳定斑块中起着重要作用。     

甘精胰岛素对人结肠癌LoVo细胞作用的研究

目的观察甘精胰岛素在体外对结肠癌LoVo细胞的影响并探讨其可能机制。方法使用不同浓度甘精胰岛素对结肠癌LoVo细胞进行干预,以人胰岛素进行对照,分别通过CCK8法和流式细胞术检测结肠癌细胞的增殖和凋亡,通过蛋白印迹法检测ERK1/2蛋白表达。结果经甘精胰岛素(20、50、100、150、200 n M)处理后LoVo细胞数量明显增加,与空白组比较,甘精胰岛素(100、150、200 n M)和阳性对照组人胰岛素(100 n M)有显著促增殖作用(P0.05)。空白组早期细胞凋亡率为(13.26±1.34)%,甘精胰岛素(100、150、200 n M)处理后凋亡率分别减少到(11.52±2.18)%、(9.11±1.39)%和(6.08±1.92)%。加入ERK1/2抑制剂PD98059后甘精胰岛素未显示出诱导LoVo细胞增殖和减少凋亡的作用。甘精胰岛素上调ERK1/2磷酸化水平并呈时间和剂量依赖性,但ERK1/2总水平没有改变。结论甘精胰岛素在体外有诱导结肠癌LoVo细胞增殖的作用,其机制可能是通过激活ERK1/2信号转导通路实现的。     

Hepatocyte growth factor/Met system promotes endometrial and endometriotic stromal cell invasion via autocrine and paracrine pathways

Endometrial stromal cells reportedly have a role in the initial invasion of endometrial tissue into the peritoneum. Hepatocyte growth factor (HGF), which is a ligand for the c-met protooncogene product (Met), stimulates proliferation and invasion of a large number of cells. In this study we investigated the role of the HGF/Met system in the pathogenesis of endometriosis. HGF concentrations in the peritoneal fluid of patients with endometriosis were significantly higher than in those without endometriosis and correlated positively with revised American Society of Reproductive Medicine scores. We showed that the peritoneum and endometriotic stromal cells may be major sources of HGF in peritoneal fluid. Endometrial and endometriotic stromal cells expressed the Met receptor, which was activated by endogenous and exogenous HGF. HGF enhanced stromal cell proliferation and invasion. We also demonstrated that the HGF-stimulated stromal cell invasion was due in part to the induction of urokinase-type plasminogen activator, a member of the extracellular proteolysis system. In conclusion, the HGF/Met system is involved in the pathogenesis of endometriosis by promoting stromal cell proliferation and invasion of shed endometria and endometrial lesions via autocrine and paracrine pathways.     

SGK1 inhibits cellular apoptosis and promotes proliferation via the MEK/ERK/p53 pathway in colitis

AIM: To investigate the role of serum-and-glucocorticoid-inducible-kinase-1(SGK1) in colitis and its potential pathological mechanisms.METHODS: SGK1 expression in mucosal biopsies from patients with active Crohn's disease(CD) and normal controls was detected by immunohistochemistry. We established an acute colitis model in mice induced by 2,4,6-trinitrobenzene sulfonicacid, and demonstrated the presence of colitis using the disease activity index, the histologic activity index and hematoxylin and eosin staining. The cellular events and potential mechanisms were implemented with small interference RNA and an inhibitor of signaling molecule(i.e., U0126) in intestinal epithelial cells(IECs). The interaction between SGK1 and the signaling molecule was assessed by coimmunoprecipitation.RESULTS: SGK1 expression was significantly increased in the inflamed epithelia of patients with active CD and TNBS-induced colitis model(0.58 ± 0.055 vs 0.85 ± 0.06, P 0.01). At the cellular level, silencing of SGK1 by small interference RNA(si SGK1) significantly inhibited the phosphorylation of mitogen-activated protein kinase kinase 1(MEK1) and the downstream molecule extracellular signal regulated protein kinase(ERK) 1/2, which induced the upregulation of p53 and Bcl-2-associated X protein, mediating the subsequent cellular apoptosis and proliferation in IECs. Cells treated with MEK1 inhibitor(i.e., U0126) before si SGK1 transfection showed a reversal of the si SGK1-induced cellular apoptosis. CONCLUSION: Our data suggested that SGK1 may protect IECs in colitis from tumor necrosis factor-α-induced apoptosis partly by triggering MEK/ERK activation.     

CD163/HO-1通路激活在稳定动脉粥样硬化斑块中的作用

动脉粥样硬化(atherosc lerosis,AS)目前被广泛认为是一种慢性炎症性疾病,炎症反应与斑块的不稳定性密切相关。CD163作为血红蛋白特异清道夫受体,近年被研究发现可以调节血红素氧合酶-1(hem e oxygenase-1,HO-1)的表达,而后者已经大量实验证明具有抗炎、抗氧化作用。因此有学者提出,CD163/HO-1通路作为一个新的抗炎途径,有望在稳定动脉粥样硬化不稳定斑块中起着重要作用。     

利拉鲁肽在体内外调节microRNA促进胰岛β细胞增殖

目的探讨利拉鲁肽对db／db小鼠胰腺及大鼠胰岛细胞瘤细胞系INS-1中microRNA表达的影响。方法将20只4周龄雄性db／db小鼠按随机数字表法分为对照组和利拉鲁肽组（每组10只）。分别给予0．1ml生理盐水或300ng／g利拉鲁肽皮下注射,每E12次。8周后测定糖化血红蛋白,行腹腔注射葡萄糖耐量试验（TPGTT）。8周末处死小鼠,免疫组化法分析小鼠胰腺B细胞增殖水平;实时荧光定量聚合酶链反应（RT-PCR）测定小鼠胰腺microRNA（miR．375和miR-34a）的表达。INS-1细胞分别给予0．5mmol/L棕榈酸培养0、48、72h或100nmol／L利拉鲁肽预处理12h后,给予0．5mmol／L棕榈酸培养72h,RT—PCR测定miR-375和miR-34a表达水平。采用t检验及方差分析进行组间数据比较分析。结果与对照组相比,利拉鲁肽组小鼠糖化血红蛋白水平降低（分别为7．3％±0．3％和4．7％±0．6％,t=16．47,P〈0．01）;IPGTT血糖曲线下面积降低[分别为（4568-4-197）和（1927±127）mmol·L-1·min-1,t=26．53,P〈0．05];胰岛素曲线下面积增加[分别为（1080±247）和（2818±378）μg·L-1·min-1,t=7．73,P〈0．05]。免疫组化法显示,与对照组相比,利拉鲁肽组小鼠胰岛素阳性面积增加（分别为1．40±0．30和0．37±0．09,t=19．14,P〈0．01）,Brdu染色阳性细胞比例增加（分别为2．40％±0．22％和0．73％±0．10％,t=4．97,P〈0．01）。利拉鲁肽组小鼠胰腺miR-375与miR-34a表达较对照组分别降低50％（分别为1．1±0．3和2．2±0．5,t=3．08,P〈0．05）和71％（分别为1．1±0．3和3．8±1．2,t=2．80,P〈0．05）。棕榈酸培养使INS-1细胞miR-375表达呈剂量、时间依赖性增加,利拉鲁肽可抑制棕榈酸诱导的miR-375表达（F=7．20,P〈0．01）。结论microRNA可能是利拉鲁肽调节β细胞增殖的靶点之-。