Identification of Halophilic Microbes in Lung Fibrotic Tissue by Oligotyping

Idiopathic pulmonary fibrosis (IPF) is an incurable disease with poor prognosis and unknown etiology. The poor clinical outcome is associated with enhanced microbial burden in bronchoalveolar lavage fluid from IPF patients. However, whether microbes from the respiratory tract fluid cause the disease remains uncertain. Tissue-associated microbes can influence host physiology in health and disease development. The aim of this study was to evaluate the existence of microbes in lung fibrotic tissues. We evaluated the microbial community in lung tissues from IPF and from human transforming growth factor-β1 (TGF-β1) transgenic mice with lung fibrosis by oligotyping. We also evaluated the microbial population in non-tumor-bearing tissues from surgical specimens of lung cancer patients. The phyla Firmicutes and the genus Clostridium tended to be predominant in the lung tissue from IPF and lung cancer patients. Oligotyping analysis revealed a predominance of bacteria belonging to the genera Halomonas, Shewanella, Christensenella, and Clostridium in lung tissue from IPF and lung cancer. Evaluation of the microbial community in the lung tissue from mice revealed abundance of Proteobacteria in both wild-type (WT) littermates and transgenic mice. However, the genus Halomonas tended to be more abundant in TGF-β1 transgenic mice compared to WT mice. In conclusion, this study describes tissue-associated microbes in lung fibrotic tissues from IPF patients and from aging TGF-β1 transgenic mice

Role of Matrix Metalloproteinase-2 in Eosinophil-Mediated Airway Remodeling

Airway remodeling is responsible for the progressive decline of lung function in bronchial asthma. Matrix metalloproteinase-2 and fibroblast-to-myofibroblast transition are involved in tissue remodeling. Here we evaluated whether eosinophils play a role in fibroblasts-to-myofibroblasts transition and in the expression of matrix metalloproteinase-2. We co-cultured human eosinophils with human fetal lung fibroblast-1 cells, assessed the expression of remodeling-associated molecules by immunoassays and polymerase-chain reaction, and eosinophils-mediated migration of human fetal lung fibroblast-1 cells using a Boyden chamber. To clarify the participation of matrix metalloproteinase-2 in airway remodeling we administered bone marrow-derived eosinophils by intra-tracheal route to transgenic mice overexpressing the human matrix metalloproteinase-2. The expression of α-smooth muscle actin significantly increased in human fetal lung fibroblast-1 cells co-cultured with human eosinophils compared to controls. There was enhanced expression of matrix metalloproteinase-2 during fibroblast-to-myofibroblast transition. An inhibitor of matrix metalloproteinases blocked eosinophils-associated fibroblast-to-myofibroblast transition and increased migration of fibroblasts. The human matrix metalloproteinase-2 transgenic mice receiving adoptive transfer of mouse eosinophils exhibited increased inflammation and advanced airway remodeling compared to wild type mice. This study demonstrated that eosinophils induce fibroblast-to-myofibroblast transition, secretion of matrix metalloproteinase-2, accelerated migration of fibroblasts, and promote matrix metalloproteinase-2-related airway remodeling. These findings provide a novel mechanistic pathway for eosinophil-associated airway remodeling in bronchial asthma

Chronic Fibrosis and Its Progression to Cancer

The terminal stage of many chronic inflammatory diseases is organ fibrosis [...

Chronic Fibrosis and Its Progression to Cancer

The terminal stage of many chronic inflammatory diseases is organ fibrosis [...

Amelioration of Diabetes by Protein S

application/pdfProtein S is an anticoagulant factor that also regulates inflammation and cell apoptosis. The effect of protein S on diabetes and its complications is unknown. This study compared the development of diabetes between wild-type and transgenic mice overexpressing human protein S and the development of diabetic glomerulosclerosis between mice treated with and without human protein S and between wild-type and protein S transgenic mice. Mice overexpressing protein S showed significant improvements in blood glucose level, glucose tolerance, insulin sensitivity, and insulin secretion compared with wild-type counterparts. Exogenous protein S improved insulin sensitivity in adipocytes, skeletal muscle, and liver cell lines in db/db mice compared with controls. Significant inhibition of apoptosis with increased expression of BIRC3 and Bcl-2 and enhanced activation of Akt/PKB was induced by protein S in islet β-cells compared with controls. Diabetic wild-type mice treated with protein S and diabetic protein S transgenic mice developed significantly less severe diabetic glomerulosclerosis than controls. Patients with type 2 diabetes had significantly lower circulating free protein S than healthy control subjects. This study shows that protein S attenuates diabetes by inhibiting apoptosis of β-cells and the development of diabetic nephropathy.本文 / Department of Diabetes, Metabolism and Endocrinology, Mie University Graduate School of Medicine12

Amelioration of Diabetes by Protein S

application/pdf内容の要旨・審査結果の要旨 / 三重大学大学院医学系研究科 生命医科学専攻 病態制御医学講座 代謝内分泌内科学分

β細胞保護を目指した新規糖尿病治療法の開発

application/pdfβ細胞の死は糖尿病の進行の原因と考えられているが、それを抑制する有用な治療法は未だ確立されていない。トロンボモジュリンは抗凝固因子として知られるが、近年、細胞保護作用を有することが注目されている。我々は糖尿病マウスにおいて、トロンボモジュリンがβ細胞を保護し、インスリン分泌を改善することにより、血糖値の上昇を抑制することを証明した。トロンボモジュリンがβ細胞保護効果を持つ新しい糖尿病治療薬として有用である可能性が示唆された。Pancreatic β-cell dysfunction is involved in the progression of diabetes. Thrombomodulin is a glycoprotein on the surface of endothelial cells which has inhibitory effect on cell apoptosis. We evaluated the effect of thrombomodulin on β-cell apoptosis and glucose tolerance in diabetic mice. Thrombomodulin attenuated hyperglycemia, ameliorated insulin secretion, and reduced β cell apoptosis. These results indicate that thrombomodulin has antidiabetic effect by suppressing β-cell apoptosis.2019年度～2020年度科学研究費補助金(若手研究)研究成果報告書19K1796

遺伝子改変マウスを用いた糸球体硬化症の新規治療戦略の開発

application/pdf本研究では、腎特異的ヒトTGFβ1過剰発現TGマウスを用いた腎線維症モデルにおいて組み換えトロンボモジュリン(rhTM)の抗線維化の効果を検討した。rhTMの投与群では、非投与群に比べ、腎組織中のコラーゲンマーカー、炎症性サイトカイン、補体系の成分の濃度が有意に低下した。rhTMの投与群では糸球体毛細血管の基底膜の肥厚、メサンギウムの拡大などは有意に改善した。ポドサイトのアポトーシスと尿細管細胞の上皮間葉系移行も有意に抑制された。In vitro実検においてrhTMはGPR15に結合し、細胞内のAktシグナル伝達経路を活性化することによって糸球体ポドサイトのアポトーシスを抑制することが判った。In present investigation, we evaluted the effect of recombinant human thrombomodulin on the kidney fibrosis and renal failure using a transgenic mouse over-expressing the complete DNA sequence of human transforming growth factor (TGF) β1 specifically in the renal glomeruli. The TGFβ1 transgenic mice treated with the recombinant human thrombomodulin showed significant decreased glomerular and tubulointerstitial deposition of extracellular matrix protein including collagen and fibronectin, reduced renal expression of pro-fibrotic and inflammatory cytokines and decreased activation of the complement system. In vitro and in vivo experiments showed that recombinant human thrombomodulin blocked apoptosis and the epithelial-mesenchymal transsition-of podocytes by binding to GPR15 and by subsequent activation of the Akt signaling pathway and blockade of Smad proteins. These results the potential of recombinant human thrombomodulin as a therapeutic drug for chronic kidney disease.2017年度～2019年度科学研究費補助金(基盤研究(C))研究成果報告書17K0844

Subcritical Water Extracts from Agaricus blazei Murrill’s Mycelium Inhibit the Expression of Immune Checkpoint Molecules and Axl Receptor

Agaricus blazei Murrill or Himematsutake is an edible and medicinal mushroom. Agaricus blazei Murrill’s fruiting body extracts have anticancer properties, although the mechanism is unknown. Basic or organic solvents, which are hazardous for human health, are generally used to prepare Agaricus blazei Murrill’s extracts. The inhibition of immune checkpoint molecules and Axl receptor is an effective therapy in cancer. This study assessed whether subcritical water extracts of the Agaricus blazei Murrill’s fruiting body or mycelium affect the expression of Axl and immune checkpoint molecules in lung cancer cells. We used A549 cells and mouse bone marrow-derived dendritic cells in the experiments. We prepared subcritical water extracts from the Agaricus blazei Murrill’s fruiting body or mycelium. The subcritical water extracts from the Agaricus blazei Murrill’s fruiting body or mycelium significantly inhibited the expression of immune checkpoint molecules and Axl compared to saline-treated cells. Additionally, the hot water extract, subcritical water extract, and the hot water extraction residue subcritical water extract from the Agaricus blazei Murrill’s mycelium significantly enhanced the expression of maturation markers in dendritic cells. These observations suggest that the subcritical water extract from Agaricus blazei Murrill’s mycelium is a promising therapeutic tool for stimulating the immune response in cancer

Antimicrobial Effects of Lactoferrin against <i>Helicobacter pylori</i> Infection

Helicobacter (H.) pylori is the primary causative agent of various gastroduodenal diseases. H. pylori is an adapted microorganism that has evolved to survive in the acidic conditions of the human stomach, possessing a natural strategy for colonizing harsh environments. Despite the implementation of various eradication regimens worldwide, the eradication rate of H. pylori has decreased to less than 80% in recent years due to the emergence of antibiotic-resistant strains. This has posed a significant challenge in treating H. pylori infection, as antibiotic resistance and side effects have become increasingly problematic. Lactoferrin, a member of the transferrin family, is an iron-binding protein with antioxidant, antibacterial, antiviral, and anti-inflammatory properties that promote human health. The concentrations of lactoferrin in the gastric juice and mucosa significantly increase during H. pylori infection and are strongly correlated with the severity of gastric mucosal inflammation. Numerous researchers have studied the antimicrobial properties of lactoferrin both in vitro and in vivo. In addition, recent studies have investigated the addition of oral lactoferrin supplementation to H. pylori eradication therapy, even though monotherapy with lactoferrin does not eradicate the microorganism. In this article, we reviewed the survival strategy of H. pylori to evade the antimicrobial activity of human lactoferrin and explore the potential of lactoferrin in H. pylori eradication therapy