全反式维甲酸对免疫细胞调控的研究进展

维生素A是人和动物为维持正常的生理功能而必须从食物中获取的一类微量有机物质。全反式维甲酸是维生素A的一种活性代谢物,它不仅调控固有免疫反应中的巨噬细胞、树突细胞和固有淋巴样细胞的功能,而且也调控获得性免疫反应中的T细胞和B细胞的功能,从而在免疫稳态中发挥重要作用。本文主要综述近年来全反式维甲酸对巨噬细胞、树突细胞、固有淋巴样细胞以及淋巴细胞调控的研究进展。     

全反式维甲酸对内皮细胞增殖和凋亡的影响及其机制

全反式维甲酸(ATRA),是体内一种重要的生物活性物质[1],以往发现在胚胎发育、抑制细胞增殖、促进细胞分化和凋亡以及视觉循环中起着重要的作用[2].有研究表明[3],其在内皮细胞的生长和塑形方面发挥调节作用,目前的研究发现ATRA做为维甲酸受体(RARs)和维甲酸X受体(RXRs)的配体可诱导碱性成纤维细胞生长因子(fibroblast geowthfactor-2,bFGF)的产生,促使内皮细胞的增殖和分化.还发现其人工合成的衍生物4-HPR具有促进内皮细胞凋亡的作用.     

全反式维甲酸对基因免疫应答的调节作用

目的　研究全反式维甲酸 (ATRA)对TR4 2 1 hCGβ质粒基因免疫诱生的特异性细胞免疫与体液免疫应答的调节作用。方法　肌肉注射重组质粒TR4 2 1 hCGβDNA(每只鼠 5 0 μg 10 0 μl)初次免疫小鼠 ,以灌胃的方式给予ATRA ,并以灌溶剂和TR4 2 1质粒免疫为对照 ;3周与 6周后经同样的方式加强免疫各组小鼠 ,采用ELISA方法对基因免疫小鼠血清中IgG抗体水平进行动态观察 ,分析小鼠血清中IgG抗体亚类 ;3H TdR掺入法测定特异性细胞增殖和细胞杀伤功能。结果　ELISA结果表明 ,TR4 2 1 hCGβ质粒基因免疫诱生较高的抗hCGβ抗体水平 ,ATRA增强TR4 2 1 hCGβ质粒基因免疫诱生的抗hCGβ特异性IgG抗体水平并且伴随IgG2a IgG1显著性降低 ;TR4 2 1 hCGβ质粒基因免疫诱生较强的淋巴细胞增殖活性和CTL活性 ,ATRA抑制TR4 2 1 hCGβ质粒基因免疫诱生的特异性细胞增殖和细胞杀伤功能。结论　ATRA促进基因免疫诱生的TH2免疫应答 ,抑制TH1型免疫应答 ,为改变基因免疫诱生的特异性免疫应答类型提供了一条新的途径。     

加替沙星-聚癸二酸酐局部缓释系统的生物相容性

背景：聚酸酐材料具有表面溶蚀性、生物可降解性和释药速率可调性,已被美国食品和药物管理局批准用于人体药物缓释材料。 目的：研究加替沙星-聚癸二酸酐局部缓释系统的制备及生物相容性。 方法：采用熔融缩聚的方法制备聚癸二酸酐,将加替沙星与聚癸二酸酐充分混合,制成载药量20%的加替沙星-聚癸二酸酐局部缓释药棒。切开6只新西兰大耳白兔背部皮肤,随机分组：实验组在脊柱两旁椎旁肌肌袋内植入聚癸二酸酐棒,对照组未植入任何材料。术后第5周取材,观察皮下组织和肌肉组织变化,以及心、肝、肾及肺的变化。 结果与结论：聚癸二酸酐生物相容性良好,植入期间兔未出现任何食欲和行为改变,植入部位未见明显红肿、出血及糜烂,植入材料表面呈多孔状,表明材料在皮下已发生降解吸收,但未发生脆裂和崩解,与周围组织形成轻微粘连;兔肝、肾、肺、心组织结构未发生改变。表明加替沙星-聚癸二酸酐局部缓释制剂无毒、无致畸作用,组织相容性好。     

全反式维甲酸可增强大鼠系膜细胞基质金属蛋白酶-2表达

一、材料与方法１．大鼠系膜细胞（ＭｓＣ）的培养与鉴定：１５０～２００ｇＳＤ大鼠购自本校动物部，取肾皮质，分离肾小球，培养与鉴定ＭｓＣ按本实验室常规方法进行［１］。实验用细胞为１０～１５代。２．Ｎｏｒｔｈｅｒｎｂｌｏｔ法检测基质金属蛋白酶２（ＭＭＰ２）ｍＲＮＡ：当ＭｓＣ长至亚融合状态后，分别在完全培养液中加入全反式维甲酸（ＡＴＲＡ，Ｓｉｇｍａ公司）１０－６、１０－７、１０－８、１０－９、０ｍｏｌ／Ｌ处理２４小时；另在亚融合状态ＭｓＣ加１０－６ｍｏｌ／Ｌ全反式维甲酸分别处理０、６、１２、２４、４…     

全反式维甲酸联合亚砷酸治疗急性早幼粒细胞白血病的临床疗效

目的探讨全反式维甲酸联合亚砷酸治疗急性早幼粒细胞白血病的临床疗效。方法选取我院在2005年03月~2013年02月收治的56例急性早幼粒细胞白血病患者随机分为观察组和对照组,观察组患者给予全反式维甲酸联合亚砷酸治疗,对照组患者给予亚砷酸治疗。结果两组患者在完全缓解率上无明显的差异性（P>0.05）,但是在达到完全缓解的时间上存在显著差异性（P<0.05）;两组患者在不良反应发生率上无明显差异性（P>0.05）。结论全反式维甲酸联合亚砷酸治疗急性早幼粒细胞白血病效果显著。     

全反式维甲酸对人乳腺癌MCF-7细胞凋亡的影响

目的:探讨全反式维甲酸(ATRA)对人乳腺癌MCF-7细胞凋亡的影响及作用途径.方法:在人乳腺癌细胞株MCF-7培养基中加入一定浓度ATRA和PKC-δ的专一抑制剂rottlerin(RO)并分组,通过SubG1assay by FACS、琼脂糖凝胶电泳检测基因组DNA ladder来观察ATRA对MCF-7的影响.结果:在浓度为5μM的ATRA作用下,MCF-7的凋亡率显著高于其它各组(P＜0.01),并可观察到明显梯状DNA.结论:ATRA能够诱导乳腺癌MCF-7细胞凋亡,但受PKC-δ的专一抑制剂RO的抑制.     

全反式维甲酸增加胃癌细胞对放射的敏感性

目的:研究全反式维甲酸(all-trans retinoic acid,ATRA)对胃癌细胞SGC-7901存活率与放射敏感性的影响,并讨论其可能的机制。方法:MTT法检测细胞存活率;平板克隆形成实验和流式细胞术分别检测细胞的放射敏感性和细胞周期;实时荧光定量PCR(RT-q PCR)检测细胞中Bax、Bcl-2、survivin与NF-κB的m RNA表达。结果:ATRA可降低SGC-7901细胞存活率,当浓度到达8μmol/L时,抑制作用达到最大;ATRA联合X射线处理后,与单纯放射处理相比,平均致死剂量(D0)和准阈剂量(Dq)显著变小(P0.05),且拟合的生存曲线明显下移;ATRA能显著降低放射诱导的细胞G_2/M期阻滞,下调SGC-7901细胞Bcl-2与survivin的m RNA表达(P0.05),上调Bax与NF-κB的m RNA表达(P0.05)。结论:ATRA能够增加胃癌细胞SGC-7901的凋亡及放射敏感性,可能与抑制细胞周期G_2/M期的阻滞作用、下调Bcl-2与survivin m RNA表达和上调NF-κB与Bax mRNA表达有关。     

全反式维甲酸对Th2极化的调节作用

为了研究全反式维甲酸(all-trans retinoic acid,ATRA)对Th细胞分化的影响和可能的机制,无菌分离BALB/c小鼠脾细胞后,用2.5μg/ml的ConA刺激脾细胞,在不同时间加入不同剂量的ATRA,72 h后收集细胞,用3H-TdR掺入法测淋巴细胞增殖活性,用RT-PCR方法检测在Th细胞分化中起作用的细胞因子的mRNA表达水平。结果ATRA呈剂量依赖抑制ConA诱导的淋巴细胞的活化,10~(-5)mol/L的ATRA对ConA诱导的淋巴细胞活化的抑制作用最强;此抑制作用与作用时间呈相关性,ConA活化淋巴细胞24 h后加入ATRA对淋巴细胞活化的抑制作用最显著,并且淋巴细胞的活化程度越低,ATRA对其的抑制作用越强。ATRA可增强经ConA活化的淋巴细胞Th2型细胞因子(IL-4、IL-6)mRNA的表达水平,而Th1型细胞因子(IFN-γ、TNF-α)mRNA的表达水平显著性降低(P<0.05)。上述结果表明ATRA总体上可抑制ConA刺激的淋巴细胞增殖活性,但却可增强,Th2细胞的分化。这一研究为Th细胞免疫偏离的基础与临床研究提供了依据。     

全反式维甲酸对小鼠肝癌细胞Hepa1-6增殖、迁移、侵袭的影响及其机制

目的探讨不同浓度的全反式维甲酸(ATRA)对小鼠肝癌细胞Hepa1-6体外增殖、凋亡、迁移和侵袭的影响及肝癌细胞间质标志蛋白和miR200家族的表达情况。方法以Hepa1-6小鼠肝癌细胞为研究对象,给予0、0.1、1.0和10.0μmol/L终浓度的ATRA处理,结晶紫染色检测细胞增殖,锥虫蓝拒染实验计数活细胞。Hoechst检测细胞凋亡,划痕实验检测迁徙能力,Transwell实验检测侵袭能力,荧光定量PCR(real-time PCR)法检测间质标志蛋白N-cadherin、sail和vimentin和0和10μmol/L ATRA处理后的miR200家族的mRNA表达。结果 ATRA处理后Hepa1-6细胞的增殖、迁移、侵袭能力明显下降(P0.05),凋亡率增高(P0.05),间质标志蛋白N-cadherin、sail和vimentin的表达明显下调(P0.05),ATRA的浓度越高,这些作用越明显。10μmol/L ATRA处理后miRNA200a-3p,200c-3p,141-3p显著上调。结论 ATRA呈浓度依赖性促进肿瘤细胞Hepa1-6凋亡,抑制其增殖、迁移及侵袭能力,这可能与ATRA上调microRNA200家族,抑制细胞的间质表型有关。     

All-trans retinoic acid up-regulates Prostaglandin-E Synthase expression in human macrophages

All-trans retinoic acid (ATRA) is a potent retinoid, which has been used successfully in different clinical settings as a potential drug to treat COPD and emphysema. In the present study, we analyzed genes modulated by ATRA by performing mRNA expression array analysis on alveolar macrophages after treatment with ATRA. Here we observed a 375-fold up-regulation of Prostaglandin-E Synthase (microsomal PGES-1, NM_004878 PTGES) which mediates the conversion of prostaglandin H(2) (PGH(2)) to Prostaglandin E(2) (PGE(2)). We furthermore studied the expression of PTGES after treatment with ATRA in human monocyte-derived macrophages (MDMs) and bronchoalveolar lavage (BAL) cells. ATRA up-regulated PTGES mRNA expression in MDMs generated with M-CSF by 2500-fold whereas in M-CSF+IL-13 macrophages the up-regulation was only 20-fold. Similarly, ATRA up-regulated PTGES mRNA expression by factor 1524 in BAL cells. The up-regulation of PTGES mRNA expression by ATRA is both time and dose dependent. IL-13 suppressed the ATRA induced PTGES expression at both mRNA and protein level in MDM and BAL cells. We also observed that LPS acts synergistically with ATRA in MDMs and strongly induces PTGES expression. ATRA had little impact on cyclooxygenase-1 and -2 (COX-1 and -2) expression as compared to PTGES expression under the same experimental conditions. Furthermore, we observed an induction of PGE(2) levels by ATRA in BAL cells. These data indicate that ATRA is a potent inducer of PTGES expression in human macrophages but not in alternatively activated macrophages and suggest that the eicosanoid pathway is important for ATRA action in macrophages.     

All-trans retinoic acid enhances bystander effect of suicide-gene therapy against medulloblastomas

In our previous study we evaluated the antitumor effect of herpes simplex virus-thymidine kinase gene (HSV-tk) on human medulloblastomas (MBs) in a therapeutic delivery system using the immortalized neural stem cell (NSC) line C17.2. However, our findings indicated that the bystander effect between C17.2tk and Daoy MB cells was weak compared to the bystander effect between NSCtk and C6 glioma cells. Gap junction intercellular communication (GJIC) is the main mechanism mediating the bystander effect in HSV-tk gene therapy. All-trans retinoic acid (ATRA) has been shown to up-regulate the expression of Connexin43 and GJIC. In this study we investigated the synergistic effect of ATRA and HSV-tk gene therapy in the treatment of MBs. We found that the expression of Connexin43 in Daoy cells was significantly increased when cells were exposed to 3 μmol/l of ATRA (P < 0.05). After co-culturing C17.2tk cells with Daoy cells at different ratios ranging from 1:1 to 1:16, ATRA significantly increased the bystander anti-tumor effect compared to ATRA-untreated cells (P < 0.05). In intracranial co-implantation experiments, mice co-implanted with C17.2tk/Daoy cells and treated with a combination of ATRA and GCV had significantly smaller tumors compared to the animals treated with GCV alone (P < 0.05). Together, our results show that ATRA enhanced the tumoricidal effect in HSVtk/GCV suicide gene therapy against Daoy MB cells by strengthening the bystander effect in vitro and in vivo.     

All-trans retinoic acid modulates fas expression and enhances chemosensitivity of human medulloblastoma cells

Retinoic acid (RA) can promote human medulloblastoma cells Med-3 toward differentiation but is not sufficient to induce cell death, suggesting its limited effect on medulloblastomas. On the other hand, the differentiated tumour cells have been supposed to be more sensitive to chemotherapeutic drugs. To elucidate this possibility for medulloblastoma cells, 10 microM/l RA, 1.0 microg/ml cisplatin (CP) and their half-dosage combinations were utilized in this study to treat Med-3 cells and their influences in cell proliferation, morphology and death patterns were evaluated. In parallel, the expressions of Fas and its ligand (FasL) were analyzed by immunocytochemical staining and Western blot hybridization. Anti-Fas antibody was used to incubate the Med-3 cells pretreated by 10 microM/l RA or 1.0 microg/ml CP. It was revealed that RA and CP could inhibit cell growth but rarely induce apoptosis. Combination of half doses each of RA and CP effectively caused most of tumour cells to die of apoptosis within 6 days. FasL molecules in 29 kDa and 37 kDa were detected in Med-3 cells with and without the treatments. The Fas molecule around 30 kDa and located in the cytoplasm was found in the normally cultured cells and the cells treated by CP. An additional 45 kDa Fas band with the appearance of its cell surface labeling was detected in the cells treated by 10 microM/l RA and by 5 microM/l RA + 0.5 microg/ml CP. The anti-Fas antibody could efficiently induce apoptosis only in the cell populations pretreated by RA. Our data thus suggest that RA can enhance the chemosensitivity of human medulloblastoma Med-3 cells presumably via modulating the Fas expression pattern. The RA/CP combined regimen would be a potential therapeutic approach for medulloblastomas.     

All-trans retinoic acid syndrome. Case report and a review of the literature

We described a patient with acute promyelocytic leukemia (APL) who developed all-trans retinoic acid syndrome (ATRAS) and reviewed the literature. ATRAS presents in patients with APL treated with all-trans retinoic acid (ATRA). It has an incidence from 5%-27% with mortality of 29%. It is secondary to ATRA effect on promyelocyte differentiation, which causes systemic inflammatory response syndrome, endothelium damage with increase in capillary permeability, microcirculation obstruction, and tissue infiltration. ATRAS clinical manifestations are fever, hypotension, respiratory, renal and hepatic insufficiency, lung infiltrates, pleural and pericardic effusion, and generalized edema. Treatment is based on ATRA suspension, support measures, and steroids.     

全反式维甲酸诱导小鼠碱性磷酸酶基因启动子区染色体重构

背景：碱性磷酸酶基因是成骨细胞分化和骨形成的重要标志。在C3H10T1/2细胞中,全反式维甲酸可通过核受体上调小鼠碱性磷酸酶的表达,与MAPK通路无关。 目的：从染色体结构调控方面揭示全反式维甲酸上调碱性磷酸酶表达的分子机制。 方法：10^-6 mol/L全反式维甲酸处理C3H10T1/2细胞0,1,6,12 h,DNA酶Ⅰ超敏感实验确定全反式维甲酸调控区域的位置,染色质免疫共沉淀实验检验全反式维甲酸处理细胞后一系列转录相关因子与全反式维甲酸调控区域结合的量效关系以及时相分布。 结果与结论：DNA聚合酶Ⅰ超敏感实验表明,小鼠碱性磷酸酶启动子转录起始位点上游约520 bp附近为潜在的全反式维甲酸调控区域;染色质免疫共沉淀实验表明,全反式维甲酸对小鼠碱性磷酸酶的上调作用是通过一系列转录相关因子的时序性共同作用来实现。表明全反式维甲酸诱导小鼠碱性磷酸酶基因转录的过程中伴随着染色质重构和组蛋白的修饰作用。     

All-trans retinoic acid-induced ectopic limb and caudal structures: murine strain sensitivities and pathogenesis.

Treatment of pregnant mice at the egg cylinder stage with retinoic acid (RA) has caused ectopic hindlimbs in the offspring. Proposed causes of ectopic hindlimbs include homeotic transformation or multiple axis formation. Two mouse strains were determined to be divergent in susceptibility to this malformation (C57BL/6N, highly sensitive; SWV/Fnn, less sensitive). Ectopic limbs were hindlimbs (expressing Pitx1 and Tbx4 but not Tbx5), yet they also expressed the predominantly forelimb Hoxb8. Ectopic body axis formation was indicated by gene expression for ectopic primitive streaks, notochords, and nodes, as well as inhibition of anterior visceral endoderm and mesodermal migration. The earlier in development that embryos were examined, the higher the rate of ectopic hindlimb development and axis formation. Ectopic axis formation and cell migration inhibition had the same strain susceptibility as the dysmorphogenesis. We propose that all extra hindlimbs were derived from ectopic axis formation, perturbation of which is genetic background dependent.     

抗Fas抗体诱导维甲酸分化后的髓母细胞瘤细胞凋亡

目的：观察全反式维甲素（ＲＡ）诱导分化前后,人髓母细胞瘤细胞系Ｍｅｄ－３对抗Ｆａｓ抗体的反应特点并探讨其免疫治疗意义。方法：使用１０μｍｏｌ／Ｌ的ＲＡ诱导Ｍｅｄ－３细胞分化并在处理得当时或第２４、４８、７２、９６ｈ,分别加入最终浓度为２５,５０,１００,２００和４００ｎｇ／ｍｌ的抗Ｆａｓ抗体,结合各组细胞Ｆａｓ表达形式,分析其形态学,细胞动力学,克隆形成能力和死亡特点。结果：正常培养的Ｍｅｄ－３细