Wnt signaling loss accelerates the appearance of neuropathological hallmarks of Alzheimer's disease in J20‐APP transgenic and wild‐type mice

Alzheimer's disease (AD ) is a neurodegenerative pathology characterized by aggregates of amyloid‐β (Aβ) and phosphorylated tau protein, synaptic dysfunction, and spatial memory impairment. The Wnt signaling pathway has several key functions in the adult brain and has been associated with AD , mainly as a neuroprotective factor against Aβ toxicity and tau phosphorylation. However, dysfunction of Wnt/β‐catenin signaling might also play a role in the onset and development of the disease. J20 APP swInd transgenic (Tg) mouse model of AD was treated i.p. with various Wnt signaling inhibitors for 10 weeks during pre‐symptomatic stages. Then, cognitive, biochemical and histochemical analyses were performed. Wnt signaling inhibitors induced severe changes in the hippocampus, including alterations in Wnt pathway components and loss of Wnt signaling function, severe cognitive deficits, increased tau phosphorylation and Aβ_1–42 peptide levels, decreased Aβ42/Aβ40 ratio and Aβ_1–42 concentration in the cerebral spinal fluid, and high levels of soluble Aβ species and synaptotoxic oligomers in the hippocampus, together with changes in the amount and size of senile plaques. More important, we also observed severe alterations in treated wild‐type (WT ) mice, including behavioral impairment, tau phosphorylation, increased Aβ_1–42 in the hippocampus, decreased Aβ_1–42 in the cerebral spinal fluid, and hippocampal dysfunction. Wnt inhibition accelerated the development of the pathology in a Tg AD mouse model and contributed to the development of Alzheimer's‐like changes in WT mice. These results indicate that Wnt signaling plays important roles in the structure and function of the adult hippocampus and suggest that inhibition of the Wnt signaling pathway is an important factor in the pathogenesis of AD .

Read the Editorial Highlight for this article on page 356 .

     

石蜡酪杆菌B126产生的糖脂的理化性质

通过纸层析、硅胶薄层层析和气相色谱分析表明,石蜡酪杆菌(Caseobacter paraffinicum)B126至少产生两种糖脂,其主要产物是海藻糖脂。它由海藻糖和两种以上的脂肪酸(十六碳酸和十八碳酸)所组成。其表面张力和界面张力(对重液体石蜡)分别为28～29和1～2mN/m,临界胶束浓度(CMC)较低,为64mg/L。     

Nobiletin suppresses cholangiocarcinoma proliferation via inhibiting GSK3β

Background: Cholangiocarcinoma (CCA) is a type of hepatobiliary cancer characterized by uncontrolled cell proliferation, with a poor prognosis and high mortality. Nobiletin (NBT) is a promising anti-tumor compound derived from the peels of oranges and other citrus plants, citrus plant. But the effect of NBT on CCA remains unknown.Results: Our data showed that NBT suppressed CCA cell proliferation in vitro and in vivo. Colony formation and Edu assay indicated that NBT inhibited cell proliferation. Cell cycle analysis showed that NBT arrested the cell cycle in G0/G1 phase. Target prediction showed that GSK3β was a direct target. Western blot and immunofluorescence confirmed that NBT reduced the phosphorylation of GSK3β. The antiproliferative effect of NBT was intercepted in GSK3β knockdown CCA cells. The cellular thermal shift assay (CETSA) showed NBT directly bound to GSK3β. Finally, NBT showed an anti-proliferative effect in tumor-bearing mice with no hepatotoxicity.Conclusion: NBT could inhibit CCA proliferation, and the pharmacological activity of NBT in CCA was attributed to its direct binding to GSK3β. We suggested that NBT might be a potential natural medicine in CCA treatment.     

Glycogen Synthase Kinase-3 modulates α1A-adrenergic receptor action and regulation

The possibility that glycogen synthase kinase 3 (GSK3) could modulate α_1A-adrenergic receptor (α_1A-AR) function and regulation was tested employing LNCaP and HEK293 cells transfected to express the enhanced green fluorescent protein-tagged human α_1A-AR. Receptor phosphorylation and internalization, intracellular free calcium, α_1A-AR-GSK3 colocalization, and coimmunoprecipitation were studied. The effects of the pharmacological GSK3 inhibitor, SB-216763, and the coexpression of a dominant-negative mutant of this kinase, as well as the signaling, desensitization, and internalization of receptors with S229, S258, S352, and S381 substitutions for alanine or aspartate, were also determined. SB-216763 inhibited agonist- and phorbol myristate acetate (PMA)-mediated α_1A-AR phosphorylation, reduced oxymetazoline-induced desensitization, and magnified that induced by PMA. Agonists and PMA increased receptor-GSK3 colocalization and coimmunoprecipitation. Expression of a dominant-negative GSK3 mutant reduced agonist- but not PMA-induced receptor internalization. α_1A-AR with the GSK3 putative target sites mutated to alanine exhibited reduced phosphorylation and internalization in response to agonists and increased PMA-induced desensitization. Agonist-induced, but not PMA-induced, receptor-β arrestin intracellular colocalization was diminished in cells expressing the GSK3 putative target sites mutated to alanine. Our data indicated that GSK3 exerts a dual action on α_1A-AR participating in agonist-mediated desensitization and internalization and avoiding PMA-induced desensitization.     

3,3',4,4',5-Pentachlorobiphenyl (PCB 126) impacts hepatic lipid peroxidation, membrane fluidity and beta-adrenoceptor kinetics in chick embryos

Polychlorinated biphenyls (PCB) and other aryl hydrocarbon receptor (AHR) agonists induce oxidative stress and alter membrane lipid peroxidation and fluidity. This study tested the hypothesis that PCB-induced changes in membrane properties impact membrane beta-adrenoceptor (beta-AR) affinity and capacity in chick embryo hepatocytes. Embryos were injected into the air cell with 1.6 microg 3,3',4,4',5-pentachlorobiphenyl (PCB 126)/kg egg at day 0, and incubated to day 19 when livers were removed. This dose resulted in hepatic PCB 126 levels of 0.67 ng/g liver or 10.2 ng/g liver lipid; levels in untreated embryos were non-detectable. Hepatic microsomal EROD activity was elevated by approximately 12-fold and embryo mortality was significantly increased compared with the untreated group. Hepatic lipid peroxidation increased and membrane order (steady-state fluorescence anisotropy values) decreased with in ovo PCB 126 exposure. Consistent with changes in membrane structure, hepatic beta-AR affinity for CGP 12177 significantly decreased (Kd increased) without changes in receptor numbers. This study demonstrates that in ovo exposure to PCB 126 in chick eggs significantly impacted embryo survival, and this was correlated with altered hepatic membrane structure and ultimately membrane function.     

IKK-β/NF-κB p65 mediates p27 protein degradation in arsenite response

p27^Kip1 is a potent inhibitor of the cyclin-dependent kinases that drive G1 to S phase transition. Since deregulation of p27^Kip1 is found in many malignancies and is associated with the poor prognosis, elucidation of the molecular bases for regulation of p27^Kip1 expression is of great significance, not only in providing insight into the understanding of biological p27^Kip1, but also in the development of new cancer therapeutic tactics. We here explored the inhibitory regulation of IKKβ on p27^Kip1 expression following arsenite exposure. We found that although the basal level of p27^Kip1 expression in the IKKβ^−/− cells is much lower than that in the IKKβ^+/+ cells, the deletion of IKKβ in the MEFs led to a marked increase in p27^Kip1 protein induction due to arsenite exposure in comparison to that in the IKKβ^+/+ cells. The IKKβ regulatory effect on p27^Kip1 expression was also verified in the IKKβ^−/− and IKKβ^−/− cells with IKKβ reconstitutional expression, IKKβ^−/− (IKKβ). Further studies indicated that IKKβ-mediated p27^Kip1 downregulation occurred at protein degradation level via p65-dependent and p50-independent manner. Moreover, the results obtained from the comparison of arsenite-induced GSK3β activation among transfectants of WT, IKKβ^−/− and IKKβ^−/− (IKKβ), and the utilization of GSKβ shRNA, demonstrated that IKKβ regulation of p27 protein degradation was mediated by GSK3β following arsenite exposure.