关键词： spermatocytes   Ahr   Nrf2   CSC   ATF3   E2F4   p38 MAPK   and ERK  

摘要： Our earlier studies have demonstrated that the cigarette smoke in the form of cigarette smoke condensate(CSC)causes growth arrest of a mouse spermatocyte cell line[GC-2spd(ts)]through activation of the AHR–NRF2 *** present study demonstrates the CSC-activated p38 and ERK MAPK signaling in GC-2spd(ts)via arylhydrocarbon receptor(AHR).Pharmacological inhibition by using AHR-antagonist,or p38 MAPK and ERK(MEK1)inhibitors significantly abrogates CSC-induced growth arrest by AHR and MAPK ***-PCR,western blot,and immunofluorescence of Ahr-target of Nrf2,and stress-inducible growth suppressive Atf3 and E2f4 following treatments indicate a crosstalk among these *** of Atf3 by Nrf2 and Ahr through RNA interference suggests the existence of a cross-regulatory loop between the *** induction of E2f4 via Atf3 and its regulation by pharmacological inhibitors reveal a possible regulatory mechanism of growth inhibitory *** silencing of Ahr,Nrf2,Atf3,and E2f4 genes and downregulation of cyclins by CSC corroborate the growth inhibitory effect of cigarette ***,the data obtained suggest that the CSC-mediated MAPKs and AHR–NRF2 crosstalks lay the molecular basis for the growth arrest and cell death of spermatocytes.

关键词： 活化转录因子3   脊髓损伤   斑马鱼模型   炎症因子   轴突再生   神经发育  

摘要： 背景：脊髓损伤是一种严重的中枢神经系统致残性疾病,目前针对脊髓损伤尚无有效的治疗手段和药物。哺乳动物的神经系统拥有非常有限的修复和再生能力。与哺乳动物不同，斑马鱼在脊髓损伤后具有很强的修复和再生能力，成年斑马鱼在脊髓损伤后六周时间内可基本恢复正常运动水平，但其具体机制尚不明确。已有研究证明活化转录因子3（ATF3）在斑马鱼视神经损伤后可以促进视神经细胞再生和修复。这提示我们，在斑马鱼脊髓损伤后，ATF3分子可能对神经再生和修复起到重要作用。\n 目的：本课题利用成年斑马鱼脊髓横断模型，探究ATF3在脊髓损伤后的mRNA和蛋白水平的变化。同时在抑制ATF3蛋白表达后，研究其对斑马鱼脊髓修复的影响和相关机制。\n 方法：选用4-6月龄的成年斑马鱼进行脊髓横断模型手术，在损伤后不同时间点取损伤点下段4mm的脊髓组织进行实时定量PCR，原位杂交以及免疫荧光，检测ATF3 mRNA和蛋白变化，同时在脊髓损伤后不同时间点用免疫荧光共标ATF3与运动神经元或ATF3与放射状胶质细胞。斑马鱼脊髓损伤后，用ATF3 morpholino（MO）抑制斑马鱼脊髓损伤后ATF3蛋白质表达，检测斑马鱼运动恢复情况以及穿过损伤位点神经轴突修复的水平变化，同时用实时定量 PCR的方法检测肿瘤坏死因子-α（TNF-α）和白介素-1β（IL-1β）mRNA的表达水平变化以探究炎症因子对修复的影响。最后，利用显微注射技术将ATF3 MO注射到斑马鱼胚胎中，探究ATF3分子对斑马鱼幼鱼发育的影响。\n 结果：\n ***3 mRNA水平在斑马鱼脊髓损伤后的4小时、12小时和6天维持在较高的表达水平，而在损伤后11天，其恢复到假手术组的表达水平。\n ***3蛋白水平在脊髓损伤后的4小时、12小时和6天表达水平升高，11天降低到假手术组水平。\n ***3蛋白表达在运动神经元的细胞浆中，并且在脊髓损伤后12小时和6天表达水平增高。\n ***3蛋白毗邻但不表达在放射状胶质细胞中，在脊髓损伤后6天，放射状胶质细胞数量增加。\n ***3 MO抑制ATF3蛋白在斑马鱼脊髓内表达后，脊髓损伤后第5和第6周，抑制组斑马鱼运动能力分别降低了36.4%和35.7%。\n 6.通过神经示踪技术发现，抑制脊髓ATF3表达六周后，神经轴突修复被抑制。\n 7.抑制脊髓ATF3表达11天后，相比MO对照组，抑制组TNF-α和IL-1βmRNA表达水平维持在高水平，在脊髓损伤后21天，恢复到对照组水平。\n ***3 MO抑制ATF3蛋白在斑马鱼胚胎发育中的表达后，幼鱼发育畸形，并且在受精5天后出现死亡率升高和身体长度变短等现象。\n 结论：ATF3分子通过调控炎症因子和轴突再生参与斑马鱼脊髓损伤后的修复，并对幼鱼的正常发育起到重要作用。

关键词： ACRYLAMIDE   MACROPHAGES   CELLULAR aging   ACTIVE oxygen -- Physiological effect   TELOMERASE   JNK mitogen-activated protein kinases   CELL cycle -- Regulation  

摘要： Senescence, which is irreversible cell cycle arrest, is induced by various types of DNA damage, including genotoxic stress. Senescent cells show dysregulation of tumor suppressor genes and other regulators of cellular proliferation. Activating transcription factor 3 (ATF3) plays a pleiotropic role in biological processes through genotoxic stress. In this study, we examined the effects of acrylamide (ACR), a genotoxic carcinogen, on cellular senescence and the molecular mechanisms of ATF3 function in macrophages. Treatment of macrophages with ACR at low concentrations (<1.0 mM) resulted in senescence-like morphology and an increase in senescence-associated beta-galactosidase (SA-beta-gal) activity. Exposure of macrophages to ACR led to stress-induced, telomerase-independent senescence. In addition, ACR treatment for 1, 3, or 5 days showed a concentration-dependent increase in ATF3 expression and G0/G1 phase arrest. To better understand the role of ATF3 in controlling the senescence response to ACR, SA-beta-gal activity was examined using ATF3 knockdown and overexpression. ACR-mediated senescence was significantly decreased by knockdown of ATF3, whereas it was increased with ATF3 overexpression. We found that ATF3 regulated p53 and p21 levels. ATF3 also played an important role in regulating intracellular reactive oxygen species (ROS) production in response to ACR treatment. Moreover, phosphorylation of p38 and JNK kinases, which were activated during ATF3-mediated senescence, was observed in ACR-treated macrophages. Taken together, these results suggest that ATF3 contributes to ACR-induced senescence by enhancing ROS production, activating p38 and JNK kinases, and promoting the ATF3-dependent expression of p53, resulting in regulation of cellular senescence in macrophages.

关键词： Basic (laboratory) research   science   translational research   science   liver transplantation   hepatology   innate immunity   liver disease: immune   inflammatory   macrophage   monocyte biology   activation  

摘要： Activating transcription factor 3 (ATF3) is a stress-induced transcription factor that has been shown to repress inflammatory gene expression in multiple cell types and diseases. However, little is known about the roles and mechanisms of ATF3 in liver ischemia/reperfusion injury (IRI). In warm and cold liver IRI models, we showed that ATF3 deficiency significantly increased ischemia/reperfusion (IR)-stressed liver injury, as evidenced by increased serum alanine aminotransferase levels, histological liver damage, and hepatocellular apoptosis. These may correlate with inhibition of the intrahepatic nuclear factor erythroid-derived 2-related factor 2/heme oxygenase-1 (NRF2/HO-1) signaling pathway leading to enhancing Toll-like receptor 4/nuclear factor kappa beta (TLR4/NF-B) activation, pro-inflammatory programs and macrophage/neutrophil trafficking, while simultaneously repressing anti-apoptotic molecules in ischemic liver. Interestingly, activation of NRF2/HO-1 signaling using an NRF2 activator, oltipraz (M2), during hepatic IRI-rescued ATF3 anti-inflammatory functions in ATF3-deficient mice. For in vitro studies, ATF3 ablation in lipopolysaccharide (LPS)-stimulated bone marrow-derived macrophages (BMMs) depressed levels of NRF2/HO-1 and PI3K/AKT, resulting in enhanced TLR4/NF-B activation. Pretreatment of LPS-stimulated BMMs with M2 increased NRF2/HO-1 expression, promoted PI3K/AKT, which in turn suppressed TLR4/NF-B-mediated proinflammatory mediators. Thus, our results first demonstrate ATF3-mediated NRF2/HO-1 signaling in the regulation of TLR4-driven inflammatory responses in IR-stressed livers. Our findings provide a rationale for a novel therapeutic strategy for managing IR-induced liver injury. This study demonstrates that activating transcription factor 3 regulates toll-like receptor 4-driven inflammatory responses by nuclear factor erythroid-derived 2-related factor 2/heme oxygenase-1, and attenuates liver ischemia reperfusion injury.

关键词： activating transcription factor-3 (ATF3)   pneumococcal infection   pneumolysin (PLY)   S. pneumoniae   Toll-like receptors (TLR)  

摘要： Activating transcription factor-3 (ATF3) acts as a negative regulator of cytokine production during Gram-negative bacterial infection. A recent study reported that ATF3 provides protection from Streptococcus pneumoniae infection by activating cytokines. However, the mechanism by which S. pneumoniae induces ATF3 after infection is still unknown. In this study, we show that ATF3 was upregulated via Toll-like receptor (TLR) pathways in response to S. pneumoniae infection in vitro. Induction was mediated by TLR4 and TLR2, which are in the TLR family. The expression of ATF3 was induced by pneumolysin (PLY), a potent pneumococcal virulence factor, via the TLR4 pathway. Furthermore, ATF3 induction is mediated by p38 mitogen-activated protein kinase (MAPK) and c-Jun N-terminal kinase (JNK). Thus, this study reveals a potential role of PLY in modulating ATF3 expression, which is required for the regulation of immune responses against pneumococcal infection in macrophages.

关键词： Inflammation   Prostaglandins   Lipid mediators  

摘要： By generating prostaglandins, cyclooxygenase-2 (Cox-2/Ptgs2) plays a critical role in regulating inflammatory responses. While several inflammatory stimuli have been shown to increase Ptgs2 expression, less is known about how the transcription of this gene is terminated. Here we show that stimulation of macrophages with yeast zymosan, a TLR2/6 and dectin-1 agonist, causes a transient increase in the expression of Ptgs2 accompanied by a simultaneous increase in the expression of the transcriptional repressor, activating transcription factor-3 (Atf3). The expression of Ptgs2 was significantly higher in resident peritoneal macrophages isolated from Atf3(-/-) mice than that from Atf3(+/+) mice and was associated with higher prostaglandin production upon stimulation with zymosan. In activated macrophages, Atf3 accumulated in the nucleus and chromatin-immunoprecipitation analysis showed that Atf3 is;recruited to the Ptgs2 promoter region. In acute peritonitis and in cutaneous wounds, there was increased leukocyte accumulation and higher levels of prostaglandins (PGE(2)/PGD(2)) in inflammatory exudates of Atf3(-/-) mice compared with WT mice. Collectively, these results demonstrate that during acute inflammation Atf3 negatively regulates Ptgs2 and therefore dysregulation of this axis could potentially contribute to aberrant Ptgs2 expression in chronic inflammatory diseases. Moreover, this axis could be a new therapeutic target for suppressing Ptgs2 expression and the resultant inflammatory responses. (C) 2015 Elsevier Inc. All rights reserved.

关键词： 转录激活因子-3   MUC5AC   香烟烟雾   激活蛋白-1   白介素-6   白介素-8  

摘要： 背景：众所周知,粘液的过表达是慢性阻塞性肺疾病(chronic obstructive pulmonary disease, COPD)的重要病理特征。研究表明,香烟烟雾(cigarette smoke,CS)暴露与COPD的发生发展关系密切。气道粘液的过度分泌可引起气道阻塞,从而导致肺功能下降。转录激活因子3(Activating transcription factor3, ATF3)是一种参与细胞反应过程的适应性反应基因。研究显示其对炎症反应具有双向调节作用,通过调节不同的靶基因从而加强或抑制炎症反应,机体会根据应激条件的性质(如应激的类型和持续的时间)和应激信号影响靶组织/细胞类型的不同而发生不同的反应。据报道,气道粘液的过表达也是气道炎症反应的一种表现形式。然而,迄今为止国内外尚未有研究探讨气道上皮细胞中ATF3的表达在调控香烟烟雾诱导的气道粘液分泌中的作用。 目的：本课题拟探讨ATF3是否参与调控COPD中MUC5AC(气道粘液的一种重要组成成分)的表达,并探索其机制。 方法：检测香烟烟雾暴露对人体气道上皮细胞HBE(正常气道上皮细胞系),NCI-H292细胞(人肺粘液表皮样癌来源细胞系)和MTEC(小鼠原代气道上皮细胞)的ATF3和MUC5AC的表达影响。利用ATF3小干扰RNA(ATF3small-interfering RNA, ATF3siRNA)和ATF3过表达质粒探讨ATF3是否与香烟烟雾诱导的MUC5AC的表达有关。利用实时定量PCR(quantitative real-time polymerase chain reaction, Q-PCR),蛋白质印迹法(Western Blot, WB)和免疫荧光(immunofluorescence, IF)检测ATF3的缺失或过表达是否影响香烟烟雾诱导的MUC5AC的表达。进一步利用免疫共沉淀(Co-Immunoprecipitation, Co-IP)及染色体免疫共沉淀(chromatin immunoprecipitation, CHIP)法验证假设,即ATF3是否通过激活蛋白-1(activator protein-1,AP-1)间接结合到MUC5AC启动子区域从而调控其表达。在体内研究中,我们用通过CSE气道滴注和熏烟制造ATF3基因敲除小鼠(ATF3-/-mice)的COPD模型,从而验证ATF3是否参与调控香烟烟雾诱导的气道粘液的表达。 结果： 1.体外实验中,香烟烟雾能显著地诱导气道上皮细胞HBE, NCI-H292和MTEC中ATF3和MUC5AC的表达。当用ATF3siRNA特异性地敲除ATF3这个基因后,香烟烟雾诱导的MUC5AC的表达显著下降,而白介素-6(interleukin-6, IL-6)和白介素-8(interleukin-8, IL-8)的表达明显增高。然而,ATF3过表达质粒未对香烟烟雾诱导的MUC5AC的表达产生明显影响。这一现象提示,ATF3可能并非直接调控MUC5AC的表达。Co-IP和CHIP实验结果证实ATF3通过AP-1信号通路作用于MUC5AC的启动子区域,从而间接调控香烟烟雾诱导的气道MUC5AC的表达。 2.体内实验中,CSE气道滴注野生型小鼠(Wild Type mice, WT mice)大气道MUC5AC的表达较对照组明显升高,然而在ATF3-/-小鼠中气道MUC5AC的表达并未明显下降。这和体内的结果并不一致。然而,在ATF3-/-小鼠模型组中,气管及血管周围中性粒细胞的浸润明显增加。(体内模型需进一步参照熏烟小鼠的结果)。 结论：本研究结果提示ATF3通过AP-1促进香烟烟雾诱导的气道MUC5AC的表达,同时ATF3能抑制香烟烟雾诱导的IL-6, IL-8的表达。ATF3在调节COPD气道粘液高分泌和炎症反应中具有重要作用,研究结果为临床的靶向治疗提供重要参考。

关键词： PGF   大鼠   黄体   MAPK   ATF3   NR4A  

摘要： 黄体的主要功能是分泌孕酮,它的退化意味着下一个发情周期的开始,因此,它在卵巢周期性调节和妊娠的维持中起着重要作用。在啮齿类动物中,黄体退化首先会发生功能性退化(孕酮合成水平明显降低),而后发生结构性退化(细胞凋亡的发生)。本论文采用苏木素-伊红染色(HE)、酶联免疫测定(ELISA)、免疫组织化学(IHC)、RT-PCR、real-time PCR、免疫印迹(Western blot)等方法,研究MAPK与立早基因(ATF3、NR4A1和NR4A2)在PGF诱导的假孕大鼠黄体退化过程中的表达模式及其可能作用,为进一步研究哺乳动物卵巢黄体退化提供试验依据。主要内容如下:***3与MAPK参与PGF诱导假孕大鼠黄体退化的作用研究为了研究ATF3与MAPK成员参与PGF诱导假孕大鼠黄体退化的作用,本试验采用PMSG/hCG联合处理构建大鼠假孕模型。在假孕第七天注射PGF,分别在0h、10 min、30 min、1h、2 h和4 h颈椎脱臼法处死大鼠。ELISA结果表明:血中孕酮水平呈时间依赖性下降。WB结果表明:在PGF处理2h,ATF3的表达量显著上升(P<0.01);而在30min,p-ERK和p-JNK表达量显著上升(P<0.01);相比之下,p-p38的表达量却没有明显变化。IHC结果表明:PGF处理后,黄体中p-ERK的染色自外向里逐渐减弱;而ATF3的染色相对均匀的分布在大、小黄体细胞的细胞核。这些结果提示PGF会诱导MAPK部分成员的磷酸化,它们的变化可能参与调控黄体退化过程中ATF3表达的升高与孕酮水平的降低。2.核受体NR4A1与NR4A2在PGF诱导的大鼠黄体退化中的定位及其表达规律通过观察核受体NR4A成员NR4A1与NR4A2在PGF诱导的大鼠黄体退化中的表达规律,推测其在大鼠黄体退化过程中的可能作用。在假孕第七天行PGF不同时间点(0 h、30 min、1 h、2 h 和 4 h)注射,采用 RT-PCR 和 real-time PCR 方法检测卵巢中NR4A1与NR4A2mRNA表达变化,IHC观察黄体中NR4A1与NR4A2的定位。结果:NR4A1蛋白主要定位在黄体细胞的细胞质中,在卵母细胞和膜细胞上也有表达,且在PGF处理后,NR4A1mRNA水平先上升而后下降,并且在30min-lh表达量最高,与生理盐水组相比差异极显著(P<0.01)。而NR4A2在黄体中呈不均匀(片状或颗粒状)分布,大部分在黄体细胞的细胞质中表达,并且在PGF处理后,NR4A2 mRNA水平的变化趋势与NR4A1保持一致,处理30 min-1 h组NR4A2mRNA水平与生理盐水组相比差异极显著(P<0.01),但NR4A2mRNA水平的变化不及NR4A1敏感。上述结果提示核受体NR4A1和NR4A2可能与PGF诱导的假孕大鼠黄体退化有密切关系。

关键词： 酒精性脂肪肝   原位灌流   kupffer细胞   ATF3   巨噬细胞极化  

摘要： 酒精性肝病(Alcoholic Hepatitis)是一种长期因大量饮酒从而导致的肝脏类疾病。通常脂肪肝为酒精性肝病的初期表现,进而可发展成酒精性肝炎、肝纤维化和肝硬化。在我国,酒精性肝病目前是常见的肝脏疾病之一,给人民健康造成了严重的危害。近年来,在同期肝病住院患者中酒精性肝病所占的比例在不断攀升,因此对酒精性肝病的研究就显得尤其重要。枯否细胞(KCs)作为定居在肝脏内的巨噬细胞,在酒精性肝病的发病过程中起到了非常重要的作用。它具有吞噬功能的同时又具有分泌功能,因此它可以利用多种途径对肝脏造成损伤。由于巨噬细胞独特的可塑性和异质性,其可根据外在环境而发生功能和表型的改变。在巨噬细胞对外界做出应答的过程中,作一个为“适应响应性”基因,细胞利用ATF-3(ATF/环磷酸腺苷(cAMP)的反应元件结合蛋白(ATF/CREB)家族的转录因子3),以适应外和/或细胞内的变化。它可以调节巨噬细胞的炎症应答的作用和相关的信号通路,如TLR4及NF-κB,来对炎症起到一个负调控的作用。1.通过建立小鼠酒精性脂肪肝的模型,并采用肝脏原位灌流,分离出形态良好、具有生物学活性的枯否细胞(KCs),研究其在酒精性脂肪肝中的表型改变。采用Lieber-DeCarli液体饮食加一次急性酒精灌胃,建立小鼠酒精性脂肪肝模型。造模周期为16天,于第16天灌胃9小时后处死小鼠,取小鼠肝脏及血清及KCs。检测各组血清谷丙转氨酶/谷草转氨酶(ALT/AST),及肝匀浆及血清中总胆固醇/三酰甘油(tg/tc)水平变化和肝脏病理切片he及油红染色。选取原位灌流的方法分离kcs,流式细胞术分析肝脏内巨噬细胞表型及组成的改变。荧光实时定量pcr(q-pcr)检测组织及提取细胞中各细胞因子水平。alt/ast、tg/tc等反应肝损伤的指标模型组显著高于对照组,he及油红染色结果与之一致,表明小鼠酒精性脂肪肝模型建立成功。肝脏原位灌流每只小鼠细胞得率约在1.5×106～2.0×106个。以小鼠巨噬细胞表面标记分子f4/80及白细胞共同抗原cd45双标设门,流式细胞术分析f4/80和cd45双阳性细胞,在模型中肝脏固有cd68+细胞显著降低,并出现大量的浸润单核细胞。q-pcr结果显示,在肝组织及原代细胞中细胞因子,如肿瘤坏死因子(tnf-α)、白介素-6(il-6)、单核细胞趋化蛋白(mcp-1)水平显著升高。小鼠酒精肝模型建立成功,肝脏原位灌流法细胞得率较高,酒精性脂肪肝的发病可能与kcs构成、表型改变,与细胞因子升高介导外周单核细胞浸润有关。2.利用ifn-γ(10ng/ml)+lps(10ng/ml)诱导raw264.7细胞6h,il-4(15ng/ml)诱导raw264.7细胞48h,在细胞上建立m1及m2极化模型。运用westernblot和q-pcr检测诱导分型后的raw264.7细胞,m1及m2各型标志性蛋白inos,arg1的表达水平。q-pcr检测各组tnf-α、tgf-β、il-6、il-10mrna表达水平,elisa检测各组细胞培养上清中il-12及il-10表达水平。westernblot和q-pcr结果显示,ifn-γ(10ng/ml)+lps(10ng/ml)刺激6h后,m1型标志性蛋白inos水平明显升高,tnf-α、il-6mrna水平也明显升高;il-4(15ng/ml)刺激48h后,arg1水平升高且tgf-β、il-10mrna表达水平也明显升高。与此同时,elisa结果显示m1型细胞上清中il-12水平明显升高,而m2型中il-10水平也明显升高。***3对于巨噬细胞极化的影响。通过westernblot检测小鼠酒精性脂肪肝模型肝组织中atf3的表达。westernblot和q-pcr检测raw264.7细胞atf3,stat3以及磷酸化stat3的表达情况,用q-pcr检测tgf-β、il-10mrna表达水平,elisa检测各组细胞培养上清中IL-12及IL-10表达水平。利用LipofectamineTM 2000将特异性ATF3转染到RAW264.7内,下调ATF3的表达,降低其活性。实验结果发现,在小鼠酒精性脂肪肝模型中肝组织ATF3的表达水平明显下降。同时在IFN-γ(10 ng/ml)+LPS(10ng/ml),诱导的M1型及IL-4(15 ng/ml)诱导的M2型RAW264.7细胞中ATF3的表达在M2型中有显著升高,磷酸化STAT3表达水平升高。转染ATF3 siRNA后,明显抑制了M2型的极化,Arg1及磷酸化STAT3表达水平均降低。

关键词： 肝再生   去迷走神经   Atf3   Nr2f1  

摘要： 目的:研究去迷走神经对大鼠再生肝中Atf3和Nr2f1的表达影响,进一步探讨迷走神经在肝再生过程中的作用及其调控机制。方法:将108只雄性大鼠分为随机的2组:PH组:2/3肝切除组;PHV组:去迷走神经2/3肝切除组。分别在术后0h、2h、12h、24h、36h、48h、72h、120h、168h处死,取大鼠的肝脏并分成两份,一份保存在中性福尔马林试剂中,另一份保存于液氮中。用免疫组织化学方法和免疫印迹法（Western Blot）检测肝组织中Atf3和Nr2f1蛋白的表达;实时荧光定量法（RT-PCR）检测肝组织中Atf3mRNA和Nr2f1 mRNA的表达。结果:（1）免疫组化法检测大鼠再生肝中Atf3的表达:PHV组大鼠在术后24h36h的阳性表达量明显高于PH组（P<0.01）,在12h和48h表达量低于PH组。（2）免疫组化法检测大鼠的再生肝中Nr2f1的表达:相对于PH组来说,PHV组大鼠的表达量在术后12h、36h、48h、72h、168h低于PH组,在术后2h、24h、120h处表达量稍高于PH组,36h表达量上升到最高,在术后12h、48h和168h时与PH组有显著性差异（P<0.05）。（3）RT-PCR法检测大鼠再生肝组织中Atf3 mRNA的表达结果:与PH组相对比,PHV组的大鼠术后24h36h肝组织内Atf3 mRNA的表达量上升,且高于PH组,在术后2h、36h和120h有显著性差异（P<0.05）。（4）RT-PCR法检测大鼠再生肝组织中Nr2f1 mRNA的表达结果:与PH组相对比,PHV组的大鼠,手术后Nr2f1 mRNA的表达量在0h168h基本都低于PH组,在术后12h和48h有极显著差异（P<0.05）。（5）WB法检测大鼠的再生肝组织中Atf3的蛋白的表达结果:与PH组相对比,PHV组大鼠手术后至48h表达量高于PH组,在48h120h表达量低于PH组,有显著性差异（P<0.05）。（6）WB法检测大鼠再生肝组织中Nr2f1的蛋白表达结果:与PH组相比,PHV组大鼠术后Nr2f1的蛋白量表达变化与PH组的变化相似,但整体都比PH组的表达量低,与PH组相比有显著性差异（P<0.01）。结论:（1）研究结果进一步提示去迷走神经可能对大鼠肝组织再生起促进作用。（2）去迷走神经对大鼠肝再生的促进作用,可能是通过促进再生肝组织中Atf3的表达,抑制再生肝组织中Nr2f1的表达来实现的。II