柞蚕杆状病毒诱导鸡异嗜白细胞脱粒效应的信号转导通路

通过探讨柞蚕(Antheraea pernyi)杆状病毒诱导鸡异嗜白细胞的信号转导途径,确证其是否具有活化鸡异嗜白细胞的作用。采用β-葡萄糖苷酸酶释放法检测柞蚕杆状病毒诱导的鸡异嗜白细胞脱颗粒反应,并通过应用蛋白酪氨酸激酶(src、syk)、磷脂酰肌醇3-激酶(PI3-K)以及丝裂原活化蛋白激酶(ERK、p38 MAPK、JNK)的特异性抑制剂,分析各蛋白酶通路在柞蚕杆状病毒诱导鸡异嗜白细胞脱粒反应中的作用。结果表明,柞蚕杆状病毒可以显著提高鸡异嗜白细胞的脱粒反应,其中src、PI3-K、JNK的特异性抑制剂能够抑制柞蚕杆状病毒诱导的鸡异嗜白细胞的脱粒反应,而syk、ERK、p38 MARK的特异性抑制剂则对鸡异嗜白细胞脱粒不起作用,说明柞蚕杆状病毒可通过src→PI3-K→JNK信号转导通路来诱导鸡异嗜白细胞的脱粒反应。     

Role of NF-kappaB in constitutive expression of MAIL in epidermal keratinocytes

Molecule possessing ankyrin-repeats induced by lipopolysaccharide (MAIL) is a nuclear IkappaB protein that is also known as interleukin-1-inducible nuclear ankyrin repeat protein and inhibitor of nuclear factor kappaBzeta (IkappaBzeta). We previously observed that MAIL-deficient mice were affected by atopic dermatitis-like skin lesions and demonstrated the importance of MAIL in the skin. In this study, we investigated MAIL expression in mouse keratinocytes. MAIL mRNA was constitutively expressed in the skin epidermis. MAIL expression was also confirmed in primary keratinocytes and the PAM212 keratinocyte cell line. The inhibitors of nuclear factor kappaB (NF-kappaB)-Bay11-7082 and the IkappaBalphaM supersuppressor-considerably downregulated MAIL expression in the keratinocytes. Immunoreactivity for NF-kappaB components was localized in the cytoplasm and nucleus of normal unstimulated keratinocytes. The expression level of MAIL in the skin did not change following lipopolysaccharide (LPS) administration to mice. Interestingly, in accordance with the in vivo findings, the MAIL expression level did not change following LPS stimulation even in primary keratinocytes; however, MAIL expression was strongly increased by interleukin-1 stimulation. These results collectively suggest that the constitutive expression of MAIL in keratinocytes is controlled, at least in part, by NF-kappaB and that there may be LPS-specific repressive mechanisms that inhibit MAIL induction.     

病毒感染与细胞核转录因子-κB信号转导的相互作用关系

自发现细胞核转录因子-κB(NF-κB)以来,人们对其分子生物学特征、作用机制及其与疾病的关系作了广泛的研究。近年来的研究结果发现,NF-κB在病毒感染与疾病发生的过程中发挥着重要作用。病毒感染真核细胞时能诱导NF-κB从细胞质转移到细胞核,从而诱导一些炎性基因转录产生大量炎性因子引起炎症反应,或阻碍细胞凋亡促进其自身在宿主细胞内的复制,或促进某些原癌基因的表达使细胞癌化等。不同病毒感染机体时,都会通过对NF-κB信号转导的影响,改变机体的某些性状,导致疾病的产生。目前已发现应激刺激、细菌脂多糖、病毒、氧自由基等很多因素能激活NF-κB,而后通过NF-κB信号转导通路的桥接作用影响机体的新陈代谢,文章就病毒感染与细胞NF-κB信号转导的相互作用关系作一综述。     

内毒素的信号转导

内毒素即脂多糖介导的细胞信号转导在激活靶细胞产生一系列应答反应，导致机体损伤并出现病理变化中起着关键作用。LPS信号转导涉及到多种血清蛋白和受体，如LBP、sCD14,mCD14，以及CD11/CD18，LBP/CD14系统在LPS信号转导中起着重要作用，但并非LPS信号转导所必需的LPS与受体结合后，细胞内信号转导涉及到酪氨酸激酶与MAPK超家族，通过多个MAPK通路和保守的三级激酶级联反应（MAPKKK-MAPKK-MAPK）将LPS信号转导致胞内，引起细胞应答，同时p38MAPK对LPS介导的细胞反应有重要调节作用，这些均表明，LPS信号转导可能是一个多受体，多通路，可调控的复杂过程。     

MAPK信号通路对脂肪细胞分化的调控

促分裂原活化的蛋白激酶(MAPK)通路主要包括胞外信号调控激酶(ERK)、p38MAPK和氨基末端蛋白激酶(JNK)三条途径,参与调节细胞增殖、分化、凋亡及细胞间的功能同步等过程,是细胞信号转导方面最为活跃的研究领域之一。研究显示MAPK也参与脂肪细胞的分化调节并发挥重要作用。ERK和p38MAPK信号通路对脂肪细胞分化的调节在不同的实验模型中表现为正调控和负调控两种不同形式;而另一成员JNK能使胰岛素受体底物1的丝氨酸发生磷酸化,进而干扰胰岛素信号,从而抑制骨髓间充质干细胞(BMSCs)的成脂分化,即对脂肪细胞分化发挥负调控作用。论文就MAPK信号通路在脂肪细胞分化中的功能进行综述,为脂类代谢性疾病的诊断和治疗提供参考。     

细胞外信号调节激酶信号途径及其与动物肌肉生长发育的关系

细胞外信号调节激酶（ＥＲＫ）是信号转导的重要分子,也是丝裂原活化蛋白激酶家族中的重要成员。ＥＲＫ信号途径是多数生长因子、细胞因子调控细胞增殖的重要途径,参与细胞分化,调控细胞的周期循环。本文总结了 ＥＲＫ信号途径的特点及其在细胞凋亡中的作用,并结合屠宰后肌肉变化的信号级联反应特点,阐述了 ＥＲＫ信号途径与动物肌肉生长发育和屠宰后肌肉变化的关系。     

茶多酚的抗氧化作用及其机制

茶多酚(TP)是一种从绿茶和红茶中提取的纯天然复合物,具有清除自由基、防止DNA受损、调节细胞内抗氧化防御系统及抗癌等生物学功能。本文系统地对TP的组成成分、抗氧化作用、对丝裂原活化蛋白激酶-核因子酶2相关因子2-抗氧化反应原件(MAPK-Nrf2-ARE)信号转导通路的调控及其相关机制进行综述。     

Molecular study on chicken tumor necrosis factor receptor-II and tumor necrosis factor receptor-associated factor-5

Tumor necrosis factor (TNF) receptor-associated factors (TRAFs) were identified as signal transducers for the tumor necrosis factor receptor (TNFR) superfamily. In this study, we cloned and characterized two genes that encode chicken TNFR-II and TRAF5. The initial cDNA fragments were obtained by suppressive subtractive hybridization (SSH) of chicken spleen cells with or without lipopolysaccharide stimulation (Salmonella typhimurium SL1181 (RE-mutant)). The results showed that chicken TNFR-II is 1518 bp in length with an open reading frame (ORF) of 1386 bp having 31% homology with human TNFR-II. Expression analysis of chicken TNFR-II revealed that it is highly expressed in the spleen and bursa of Fabricius. The chicken cell lines IN24, MSB1 and 1104B express TNFR-II abundantly. The time course analysis of expression in spleen, bursa of Fabricius and IN24 cell line showed that TNFR-II is maximally expressed at 6 h after stimulation in bursa of Fabricius and after 8 h stimulation in the IN24 cell line. With regard to TRAF5, the complete sequence was 1936 bp in length with an ORF of 1671 bp that showed 71.3% homology with human TRAF5. Expression analysis showed that, among the tissues examined, TRAF5 was strongly expressed in spleen and bursa of Fabricius, while among the cell lines examined, it was maximally expressed in IN24. Thus, both genes were expressed in the same tissues and cell line among examined materials. These results suggest that chicken TNFR-II may interact with TRAF5 adaptor protein to complete its signal transduction pathway.     

Differential responses of macrophages to Salmonella enterica serovars Enteritidis and Typhimurium

Macrophages are major effectors against Salmonella infection, and also transport bacteria between host tissues and provide a protected site for intracellular bacterial replication. We hypothesized that differences in chicken macrophage responses to Salmonella enterica serovar Enteritidis (SE) and serovar Typhimurium (ST) played a role in preferential infection of eggs by SE compared with ST. To test this hypothesis, we determined bacterial phagocytosis and intracellular viability and macrophage nitric oxide (NO) production following in vitro infection with SE or ST in the presence or absence of interferon-gamma (IFN-gamma). The effects of bacterial components, lipopolysaccharide (LPS), outer membrane proteins (OMP) and flagella, on NO production were also assessed. Our results showed: (1) in the presence or absence of IFN-gamma, the percentage macrophages phagocytizing SE and ST was similar; (2) the number of intracellular viable SE was significantly reduced compared with ST in the presence or absence of IFN-gamma; (3) increased macrophage necrosis was seen in the presence of IFN-gamma and ST; (4) Salmonella infection acted synergistically with IFN-gamma in induction of nitric oxide production; and (5) in the absence of IFN-gamma, macrophages produced significantly greater NO following treatment with SE outer membrane protein or flagella compared with ST OMP or flagella, while in the presence of IFN-gamma significantly less NO was produced following treatment with SE-LPS compared with ST-LPS. These results suggest that differential responses of chicken macrophages to SE versus ST may result in increased macrophage death with ST, which could result in an increased inflammatory response as compared to SE.     

黄芪多糖对小鼠免疫细胞信号转导相关分子的影响

为探讨黄芪多糖(APS)免疫调节的作用机理，观察了黄芪多糖对小鼠腹腔巨噬细胞一氧化氮(NO)生成、对体外培养的小鼠脾脏淋巴细胞内游离钙离子水平和蛋白激酶C(PKC)活性的影响。结果显示黄芪多糖能明显促进小白鼠腹腔巨噬细胞NO生成，显著升高小鼠脾淋巴细胞内游离钙离子的水平，引起细胞PKC活性明显升高。提示黄芪多糖通过NO介导信号传递通路，调节脾淋巴细胞内游离钙离子的浓度，升高细胞蛋白激酶活性而影响机体免疫细胞的信号转导，发挥其免疫调节作用。     

Reduced nitric oxide production and iNOS mRNA expression in IFN-gamma-stimulated chicken macrophages transfected with iNOS siRNAs

Utilizing RNA interference technology with siRNA in the HD11 macrophage cell line, we determined how the inhibition or knock-down of the iNOS (inducible nitric oxide synthase) gene affected IFN-gamma-induced macrophage production of nitric oxide (NO) and mRNA expression of genes involved in this biological pathway in the chicken. Chicken macrophages produce NO when stimulated with recombinant chicken IFN-gamma, however, when transfected with iNOS siRNAs, the production of NO is significantly decreased. We observed a 14-28% reduction in NO production by IFN-gamma-stimulated HD11 cells at 48h after initial siRNA transfection compared to non-transfected IFN-gamma-stimulated macrophages. Significant knock-down of iNOS mRNA expression (15 to 50-fold lower) was observed for each of four iNOS siRNAs, when compared to non-transfected IFN-gamma-stimulated macrophages and to those treated with a negative control siRNA. The IFN-gamma-stimulated chicken macrophages transfected with iNOS siRNAs did not show altered levels of mRNA expression for genes involved in IFN-gamma signaling and iNOS pathways (IL-1beta, IL-6, IFN-gamma, TGF-beta4, or SOCS-3) suggesting that the observed decrease in NO production is a direct result of siRNA mediated knock-down of iNOS, rather than IFN-gamma-induced changes in the other genes tested.     

Biphasic activation of PI3K/Akt and MAPK/Erk1/2 signaling pathways in bovine herpesvirus type 1 infection of MDBK cells

ABSTRACT: Many viruses have been known to control key cellular signaling pathways to facilitate the virus infection. The possible involvement of signaling pathways in bovine herpesvirus type 1 (BoHV-1) infection is unknown. This study indicated that infection of MDBK cells with BoHV-1 induced an early-stage transient and a late-stage sustained activation of both phosphatidylinositol 3-kinase (PI3K)/Akt and mitogen activated protein kinases/extracellular signal-regulated kinase 1/2 (MAPK/Erk1/2) signaling pathways. Analysis with the stimulation of UV-irradiated virus indicated that the virus binding and/or entry process was enough to trigger the early phase activations, while the late phase activations were viral protein expression dependent. Biphasic activation of both pathways was suppressed by the selective inhibitor, Ly294002 for PI3K and U0126 for MAPK kinase (MEK1/2), respectively. Furthermore, treatment of MDBK cells with Ly294002 caused a 1.5-log reduction in virus titer, while U0126 had little effect on the virus production. In addition, the inhibition effect of Ly294002 mainly occurred at the post-entry stage of the virus replication cycle. This revealed for the first time that BoHV-1 actively induced both PI3K/Akt and MAPK/Erk1/2 signaling pathways, and the activation of PI3K was important for fully efficient replication, especially for the post-entry stage.     

MAPK信号通路在动物繁殖中的研究进展

在细胞中存在着各种信号转导通路,不同的信号转导通路相互交叉,形成复杂的信号转导网络系统,参与生物体内的各种生理生化功能,其中部分信号转导通路对动物的生殖生理有着重要作用。丝裂原活化蛋白激酶(mitogen-activated protein kinases,MAPKs)是细胞内重要的信号通路之一,其参与了多种生理过程的调节,对动物的繁殖有重要作用。文章将主要对MAPK信号通路在动物繁殖中的作用进行综述。     

Research Advance on MAPK Signaling Pathways in Animal Reproduction

There were various signal transduction pathways in cells.The interaction of different signaling pathways formed a complex signal transduction system, which involved in various physiological and biochemical functions of animals, and part of them played an important role in the animal reproductive physiology.Mitogen-activated protein kinases was one of the important signaling pathways in cells, which involved in the regulation of a variety of physiological processes.Especailly, they played an important role in animal reproduction.The article will summarize the roles of MAPK signaling pathways in animal reproduction.     

Mechanisms of the Eimeria tenella growth inhibitory activity induced by concanavalin A and reticuloendotheliosis virus supernatants with interferon gamma activity in chicken macrophages and fibroblasts

Pretreatment of chicken bone marrow macrophages and embryo fibroblasts with supernatants containing chicken interferon gamma (IFN-gamma) for 24 hr prior to inoculation inhibited intracellular Eimeria tenella replication, measured by [3H] uracil incorporation. The supernatants (Sns) were obtained from culture of lymphoblastoid cells transformed by a reticuloendotheliosis virus (REV) and chicken splenocytes stimulated with concanavalin A (Con A). The mechanisms of the E. tenella growth inhibitory activity induced by Sn REV and Sn Con A in chicken macrophages and fibroblasts were studied. Addition of oxygen scavengers (superoxide dismutase, D-mannitol, DABCO, benzoic acid, L-histidine hydrochloride) was able to overcome the inhibition of E. tenella replication after pretreatment with Sn REV or Sn Con A in macrophage cultures but not in fibroblast cultures. Nitric oxide (NO) synthesis was induced in macrophage culture treated with Sn REV or Sn Con A but not in fibroblast culture. Addition of NG monomethyl-L-arginine, an NO synthase inhibitor together with the supernatants was also able to overcome inhibition of E. tenella replication in macrophage culture. On the other hand, addition of L-tryptophan to Sn REV- or Sn Con A-treated fibroblasts was able to reverse the inhibitory effect on E. tenella replication. In conclusion, production of inorganic NO or toxic oxygen intermediates may be involved in the E. tenella growth inhibitory activity of chicken macrophages pretreated with supernatants containing an IFN-gamma activity, and cellular tryptophan depletion may be involved for chicken fibroblasts, thus matching the mechanisms of the IFN-gamma-induced growth inhibitory activity for protozoans in mammals.     

The membrane‐type estrogen receptor G‐protein‐coupled estrogen receptor suppresses lipopolysaccharide‐induced interleukin 6 via inhibition of nuclear factor‐kappa B pathway in murine macrophage cells

The female sex hormone estrogen exerts anti‐inflammatory effects. The G‐protein‐coupled estrogen receptor (GPER) has been recently identified as a novel membrane‐type estrogen receptor that can mediate non‐genomic estrogenic effects on many cell types. We previously demonstrated that GPER inhibits tumor necrosis factor alpha‐induced expression of interleukin 6 (IL‐6) through repression of nuclear factor‐kappa B (NF‐κB) promoter activity using human breast cancer cells. Although several reports have indicated that GPER suppresses Toll‐like receptor‐induced inflammatory cytokine expression in macrophages, the molecular mechanisms of the inhibition of cytokine production via GPER remain poorly understood. In the present study, we examined GPER‐mediated inhibition of IL‐6 expression induced by lipopolysaccharide (LPS) stimulation in a mouse macrophage cell line. We found that the GPER agonist G‐1 inhibited LPS‐induced IL‐6 expression in macrophage cells, and this inhibition was due to the repression of NF‐κB promoter activity by GPER. G‐1 treatment also decreased the phosphorylation of inhibitor of κB kinases. Among the mitogen‐activated protein kinases, the phosphorylation of c‐jun N‐terminal kinase (JNK) was increased by G‐1. These findings delineate the novel mechanism of the inhibition of LPS‐induced IL‐6 through GPER‐activated JNK‐mediated negative regulation of the NF‐κB pathway in murine macrophage cells, which links anti‐inflammatory effects to estrogen.     

小型猪特异性麻醉颉颃剂对大鼠脑NO/cGMP信号转导系统的动态影响

为探讨NO/cGMP信号转导系统对小型猪特异性麻醉颉颃剂催醒分子机理的调控,144只Wistar大鼠随机分为对照组和试验组,对照组和试验组再分为早期催醒组、中期催醒组和晚期催醒组。用分光光度法测定脑NOS活性和NO产量,ELISA测定脑cGMP含量。结果显示,小型猪特异性麻醉颉颃剂能明显地激活大鼠相关脑区的NOS活性和NO及cGMP产量,并且大鼠不同脑区NOS活性、NO产量和cGMP含量的变化趋势不仅与大鼠行为学变化相平行,还与不同时期表现出的催醒效果基本吻合。这表明小型猪特异性麻醉颉颃剂的催醒可能与激活特定脑区的NO/cGMP信号转导系统相关。