Lung Epithelial TRPA1 Mediates Lipopolysaccharide-Induced Lung Inflammation in Bronchial Epithelial Cells and Mice

Toll-like receptor (TLR) 4 was originally thought to be the sole pattern recognition receptor for lipopolysaccharide (LPS). Transient receptor potential ankyrin 1 (TRPA1), a Ca2+-permeant channel, has been suggested as a non-TLR receptor membrane-bound sensor of LPS. We recently reported that TRPA1 is expressed in lung epithelial cells (LECs) and mediates lung inflammation induced by cigarette smoke. However, the role of TRPA1 in LPS-induced lung inflammation has not been conclusively defined, and its underlying cellular mechanisms remain unclear. In this study, our in vitro results showed that LPS sequentially produced a cascade of events, including the elevation of intracellular Ca2+, the activation of NADPH oxidase, increase in intracellular reactive oxygen species (ROS), the activation of mitogen-activated protein kinase (MAPK)/nuclear factor-kB (NF-κB) signaling, and the induction of IL-8. The increase in intracellular Ca2+ was inhibited by HC030031 (a TRPA1 antagonist) but was unaffected by TAK-242 (a TLR-4 inhibitor). The activation of NADPH oxidase was prevented by its inhibitor apocynin, EGTA (an extracellular Ca2+ chelator), and HC030031. The increase in intracellular ROS was attenuated by apocynin, N-acetyl-cysteine (NAC, a ROS scavenger), EGTA, and HC030031. The activation of the MAPK/NF-κB signaling was halted by NAC, EGTA, and HC030031. IL-8 induction was suppressed by HC030031 and TRPA1 siRNA, and further reduced by the combination of HC030031 and TAK-242. Our in vivo studies showed that trpa1–/– mice exhibited a reduced level of LPS-induced lung inflammation compared with wild-type mice as evidenced by the alleviations of increases in vascular permeability, inflammatory cell infiltration, inflammatory cytokine levels, oxidative stress, and MAPK signaling activation. Thus, in LECs, LPS may activate TRPA1 resulting in an increase in Ca2+ influx. The increased intracellular Ca2+ leads to NADPH oxidase activation, which causes an increase in intracellular ROS. The intracellular ROS activates the MAPK/NF-κB signaling resulting in IL-8 induction. This mechanism may possibly be at work to induce lung inflammation in mice

Atypical Antipsychotic Drug Olanzapine Deregulates Hepatic Lipid Metabolism and Aortic Inflammation and Aggravates Atherosclerosis

Background/Aims: Olanzapine, an atypical antipsychotic drug, has therapeutic effects for schizophrenia. However, clinical reports indicate that patients taking atypical antipsychotic drugs are at high risk of metabolic syndrome with unclear mechanisms. We investigated the effect of olanzapine on atherosclerosis and the mechanisms in apolipoprotein E-null (apoE-/-) mice. Methods: ApoE-/- mice were used as in vivo models. Western blot analysis was used to evaluate protein expression. Conventional assay kits were applied to assess the levels of cholesterol, triglycerides, free cholesterol, cholesteryl ester, fatty acids, glycerol, and cytokines. Results: Daily treatment with olanzapine (3 mg/kg body weight) for four weeks increased mean arterial blood pressure and the whitening of brown adipose tissue in mice. In addition, olanzapine impaired aortic cholesterol homeostasis and exacerbated hyperlipidemia and aortic inflammation, which accelerated atherosclerosis in mice. Moreover, lipid accumulation in liver, particularly total cholesterol, free cholesterol, fatty acids, and glycerol, was increased with olanzapine treatment in apoE-/- mice by upregulating the expression of de novo lipid synthesis-related proteins and downregulating that of cholesterol clearance- or very low-density lipoprotein secretion-related proteins. Conclusion: Olanzapine may exacerbate atherosclerosis by deregulating hepatic lipid metabolism and worsening hyperlipidemia and aortic inflammation

Isolation and charactization of very virluent Marek's disease virus in Taiwan and establishment of its vaccination program

採集自疑似雞馬立克氏病(Marek's disease,MD)病雞血液及脾臟病材 , 接種於雞胚胎纖維芽母細胞及小雞腹腔,共得到 7 株馬立克氏病毒( Marek's disease virus,MDV)。由 MDV 感染細胞之病變細胞,瓊脂沉降反 應,雞胚胎漿尿膜病變,及螢光抗體染色法結果,7株MDV可分成血清型共三 個 ,血清型1型的是LTB-1,LTB-2,LTB-3,LTS-1,LTS-2等5株,2型的LTB-4,3 型的 LTB-5。選取1型的LTB-1,LTS-1,Md/5株接種於肉雞,蛋雞和六種臺灣 土雞的實驗雞,可見實驗雞表現MD症狀和病變及免疫器官明顯萎縮,而經1 日齡HVT免疫接種,11日齡LTB-1攻毒接種後,仍然造成免疫雞之腫瘍病變及 早期斃死;根據病原性及HVT疫苗保護性試驗的結果,LTB-1,LTS-1株應屬於 vvMDV。由一日齡接種三種血清型苗的多價疫苗保護性試驗之結果顯示,二 價與三價疫苗的保護效果比單價疫苗好。Inocula from blood and spleen of Marek's disease (MD) chickens were inoculated into chicken embryo fibroblast (CEF) and abominal cavity of day-old chicks. Seven isolates of Marek's diease virus (MDV) were isolated during the experiment.By the results of cyto- pathic effect, agar gel precipitation and fluorescent antibody tests , 7 isolates were divided into 3 serotypes : serotye 1 of strains LTB-1, LTB-2, LTB-3, LTS-1, LTS-2 ; serotype 2 of strains LTB-4; serotype 3 of strain LTB-5. Strain LTB-1, LTS-1 and Md/5 of the serotype 1 were used to inoculate into the experimental chickens of layers, broilers and six lines of Taiwan local chickens. By the results of pathogenic tests and protection tests of HVT vaccine , strain LTB-1, LTS-1 were considered to be the very virluent MDV (vvMDV). From the results of protection experiments, chickens were ino- culated by monovalent or polyvalent vaccine at 1 day- old,bivalent and trivalent vaccine showed highest efficacy

β Common Receptor Mediates Erythropoietin-Conferred Protection on OxLDL-Induced Lipid Accumulation and Inflammation in Macrophages

Erythropoietin (EPO), the key factor for erythropoiesis, also protects macrophage foam cells from lipid accumulation, yet the definitive mechanisms are not fully understood. β common receptor (βCR) plays a crucial role in the nonhematopoietic effects of EPO. In the current study, we investigated the role of βCR in EPO-mediated protection in macrophages against oxidized low-density lipoprotein- (oxLDL-) induced deregulation of lipid metabolism and inflammation. Here, we show that βCR expression was mainly in foamy macrophages of atherosclerotic aortas from apolipoprotein E-deficient mice. Results of confocal microscopy and immunoprecipitation analyses revealed that βCR was colocalized and interacted with EPO receptor (EPOR) in macrophages. Inhibition of βCR activation by neutralizing antibody or small interfering RNA (siRNA) abolished the EPO-conferred protection in oxLDL-induced lipid accumulation. Furthermore, EPO-promoted cholesterol efflux and upregulation of ATP-binding cassette (ABC) transporters ABCA1 and ABCG1 were prevented by pretreatment with βCR neutralizing antibody or βCR siRNA. Additionally, blockage of βCR abrogated the EPO-conferred anti-inflammatory action on oxLDL-induced production of macrophage inflammatory protein-2. Collectively, our findings suggest that βCR may play an important role in the beneficial effects of EPO against oxLDL-elicited dysfunction of macrophage foam cells