Milk-derived exosomes (MDEs) have a different biological effect on normal fetal colon epithelial cells compared to colon tumor cells in a miRNA-dependent manner

Abstract Background Breastfeeding is the ideal source of infant nutrition. Human milk consists not only of nutrients but also biologically active components. Among these latter compounds, exosomes contain proteins, lipids, mRNAs and miRNAs. Methods To elucidate the biological effects of milk-derived exosomes (MDEs) on normal colonic epithelial cells compared to colonic tumor cells, we incubated cells with MDEs. MDEs were able to enter into normal and tumor cells and change their miRNA expression profiles. Proliferation, cell morphology and protein expression were analyzed in these cells. Results Human milk-derived exosomes induced proliferation- and epithelial mesenchymal transformation-related changes, such as collagen type I and twist expression, in normal but not in tumor cells. PTEN, a target of miRNA-148a, was downregulated in normal but not in tumor cells following incubation with MDEs. Moreover, miRNA-148a-3p knockdown cells were used to demonstrate the importance of miRNA in the effect of exosomes on cell proliferation and protein expression. MDEs inhibited proliferation and DNMT1 expression in cells with knockdown of miRNA-148a. Conclusions In conclusion, the positive effect of exosomes on normal cells without affecting tumor cells may presents an aspect of their safety when considering it use as a nutritional supplement to infant formula.

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MicroRNA-132-3p suppresses type I IFN response through targeting IRF1 to facilitate H1N1 influenza A virus infection

Bioscience Reports ◽

10.1042/bsr20192769 ◽

2019 ◽

Vol 39 (12) ◽

Author(s):

Fangyi Zhang ◽

Xuefeng Lin ◽

Xiaodong Yang ◽

Guangjian Lu ◽

Qunmei Zhang ◽

...

Keyword(s):

Influenza A Virus ◽

Peripheral Blood ◽

Influenza A ◽

Expression Profiles ◽

Protein A ◽

H1n1 Influenza ◽

Type I ◽

Dependent Manner ◽

Type I Ifn

Abstract Increasing evidence has indicated that microRNAs (miRNAs) have essential roles in innate immune responses to various viral infections; however, the role of miRNAs in H1N1 influenza A virus (IAV) infection is still unclear. The present study aimed to elucidate the role and mechanism of miRNAs in IAV replication in vitro. Using a microarray assay, we analyzed the expression profiles of miRNAs in peripheral blood from IAV patients. It was found that miR-132-3p was significantly up-regulated in peripheral blood samples from IAV patients. It was also observed that IAV infection up-regulated the expression of miR-132-3p in a dose- and time-dependent manner. Subsequently, we investigated miR-132-3p function and found that up-regulation of miR-132-3p promoted IAV replication, whereas knockdown of miR-132-3p repressed replication. Meanwhile, overexpression of miR-132-3p could inhibit IAV triggered INF-α and INF-β production and IFN-stimulated gene (ISG) expression, including myxovirus protein A (MxA), 2′,5′-oligoadenylate synthetases (OAS), and double-stranded RNA-dependent protein kinase (PKR), while inhibition of miR-132-3p enhanced IAV triggered these effects. Of note, interferon regulatory factor 1 (IRF1), a well-known regulator of the type I IFN response, was identified as a direct target of miR-132-3p during HIN1 IAV infection. Furthermore, knockdown of IRF1 by si-IRF1 reversed the promoting effects of miR-132-3p inhibition on type I IFN response. Taken together, up-regulation of miR-132-3p promotes IAV replication by suppressing type I IFN response through its target gene IRF1, suggesting that miR-132-3p could represent a novel potential therapeutic target of IAV treatment.

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Interferon-γ inhibits sirtuin 6 gene expression in intestinal epithelial cells through a microRNA-92b-dependent mechanism

AJP Cell Physiology ◽

10.1152/ajpcell.00335.2019 ◽

2020 ◽

Vol 318 (4) ◽

pp. C732-C739

Author(s):

Fangyi Liu ◽

Xiao Wang ◽

Hua Geng ◽

Heng-Fu Bu ◽

Peng Wang ◽

...

Keyword(s):

Epithelial Cells ◽

Protein Expression ◽

Intestinal Epithelial Cells ◽

Specific Inhibitor ◽

In Silico Analysis ◽

Interferon Γ ◽

Luciferase Assay ◽

Epithelial Cell Line ◽

Dependent Manner ◽

Intestinal Epithelial

Sirtuin 6 (Sirt6) is predominantly expressed in epithelial cells in intestinal crypts. It plays an important role in protecting intestinal epithelial cells against inflammatory injury. Previously, we found that colitis is associated with the downregulation of Sirt6 protein in the intestines. Here, we report that murine interferon-γ (Ifnγ) inhibits Sirt6 protein but not mRNA expression in young adult mouse colonocytes (YAMC, a mouse colonic epithelial cell line) in a dose- and time-dependent manner. Using microRNA array analysis, we showed that Ifnγ induces expression of miR-92b in YAMC cells. With in silico analysis, we found that the Sirt6 3′-untranslated region (UTR) contains a putative binding site for miR-92b. Luciferase assay showed that Ifnγ inhibited Sirt6 3′-UTR activity and this effect was mimicked by miR-92b via directly targeting the miR-92b seed site in the 3′-UTR of Sirt6 mRNA. Furthermore, Western blot demonstrated that miR-92b downregulated Sirt6 protein expression in YAMC cells. Blocking miR-92b with a specific inhibitor attenuated the inhibitory effect of Ifnγ on Sirt6 protein expression in the cells. Collectively, our data suggest that Ifnγ inhibits Sirt6 protein expression in intestinal epithelial cells via a miR-92b-mediated mechanism. miR-92b may be a novel therapeutic target for rescuing Sirt6 protein levels in intestinal epithelial cells, thereby protecting against intestinal mucosal injury caused by inflammation.

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Skin Regeneration Effect and Chemical Composition of Essential Oil from Artemisia montana

Natural Product Communications ◽

10.1177/1934578x1400901123 ◽

2014 ◽

Vol 9 (11) ◽

pp. 1934578X1400901 ◽

Cited By ~ 2

Author(s):

Mi-So Yoon ◽

Kyung-Jong Won ◽

Do Yoon Kim ◽

Dae il Hwang ◽

Seok Won Yoon ◽

...

Keyword(s):

Essential Oil ◽

Human Skin ◽

Type Iv Collagen ◽

Steam Distillation ◽

Biological Effects ◽

Skin Regeneration ◽

Type I ◽

Dependent Manner ◽

Positive Effects ◽

Type Iv

Artemisia montana Pampan (Compositae) (AMP) contains various compounds, including phenolic acids, alkaloids, and essential oil. It has been widely used in oriental medicine due to a variety of biological effects. However, the biological activity of the essential oil from AMP (AMPEO) on skin has not been investigated. In the present study, AMPEO was evaluated for its composition and its effect on cellular events (migration and proliferation) related to skin regeneration using normal human keratinocytes (HaCats). AMPEO, which was extracted by steam distillation, contained 42 components. AMPEO increased proliferation in HaCats in a dose-dependent manner (EC 50, 8.5 ng/mL) and did not affect migration. AMPEO also enhanced the phosphorylation of Akt and ERK 1/2 and induced the synthesis of type IV collagen, but not type I collagen in HaCats. In addition, AMPEO promoted wound closure in the dorsal side skin of rat tail. These results demonstrated that AMPEO extracted by steam distillation induced proliferation and synthesis of type IV collagen in human skin keratinocytes, and may thereby exert positive effects on skin regeneration and wound healing in human skin.

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Altered expression of p63 isoforms and expansion of p63- and club cell secretory protein-positive epithelial cells in the lung as novel features of aging

AJP Cell Physiology ◽

10.1152/ajpcell.00330.2018 ◽

2019 ◽

Vol 316 (4) ◽

pp. C492-C508 ◽

Cited By ~ 2

Author(s):

Jutaro Fukumoto ◽

Sahebgowda Sidramagowda Patil ◽

Sudarshan Krishnamurthy ◽

Smita Saji ◽

Irene John ◽

...

Keyword(s):

Epithelial Cells ◽

Protein Expression ◽

Basal Cell ◽

Expression Profiles ◽

Type Ii ◽

Cell Marker ◽

Altered Expression ◽

Club Cell ◽

Protein Expression Profiles ◽

Old Mice

Aging is a key contributor for subclinical progression of late-onset lung diseases. Basal, club, and type II alveolar epithelial cells (AECs) are lung epithelial progenitors whose capacities of differentiation are extensively studied. The timely transition of these cells in response to environmental changes helps maintain the intricate organization of lung structure. However, it remains unclear how aging affects their behavior. This paper demonstrates that the protein expression profiles of a type II AEC marker, prosurfactant protein C (pro-SPC), and a basal cell marker, p63, are altered in the lungs of 14-mo-old versus 7- to 9-wk-old mice. Expression of NH2-terminal-truncated forms of p63 (ΔNp63), a basal cell marker, and claudin-10, a club cell marker, in cytoplasmic extracts of lungs of 14-mo-old mice was upregulated. In contrast, nuclear expression of full-length forms of p63 (TAp63) decreases with age. These alterations in protein expression profiles coincide with dramatic changes in lung functions including compliance. Whole tissue lysates of middle-aged versus aged rhesus monkey lungs display similar age-associated alterations in pro-SPC expression. An age-associated decrease of TAp63 in nuclear lysates was observed in aged monkey group. Moreover, the lungs of 14-mo-old versus 7- to 9-wk-old mice display a wider spreading of ΔNp63-positive CCSP-positive bronchiolar epithelial cells. This expansion did not involve upregulation of Ki67, a representative proliferation marker. Collectively, it is postulated that 1) this expansion is secondary to a transition of progenitor cells committed to club cells from ΔNp63-negative to ΔNp63-positive status, and 2) high levels of cytoplasmic ΔNp63 expression trigger club cell migration.

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Isopsoralen Enhanced Osteogenesis by Targeting AhR/ERα

Molecules ◽

10.3390/molecules23102600 ◽

2018 ◽

Vol 23 (10) ◽

pp. 2600 ◽

Cited By ~ 8

Author(s):

Luna Ge ◽

Yazhou Cui ◽

Kai Cheng ◽

Jinxiang Han

Keyword(s):

Osteoblast Differentiation ◽

Estrogen Receptor Alpha ◽

Biological Effects ◽

Hormone Deficiency ◽

In Vivo Studies ◽

Osteogenic Potential ◽

Type I ◽

Dependent Manner

Isopsoralen (IPRN), one of the main effective ingredients in Psoralea corylifolia Linn, has a variety of biological effects, including antiosteoporotic effects. In vivo studies show that IPRN can increase bone strength and trabecular bone microstructure in a sex hormone deficiency-induced osteoporosis model. However, the mechanism underlying this osteogenic potential has not been investigated in detail. In the present study, we investigated the molecular mechanism of IPRN-induced osteogenesis in MC3T3-E1 cells. Isopsoralen promoted osteoblast differentiation and mineralization, increased calcium nodule levels and alkaline phosphatase (ALP) activity and upregulated osteoblast markers, including ALP, runt-related transcription factor 2 (RUNX2), and collagen type I alpha 1 chain (COL1A1). Furthermore, IPRN limited the nucleocytoplasmic shuttling of aryl hydrocarbon receptor (AhR) by directly binding to AhR. The AhR target gene cytochrome P450 family 1 subfamily A member 1 (CYP1A1) was also inhibited in vitro and in vivo. This effect was inhibited by the AhR agonists indole-3-carbinol (I3C) and 3-methylcholanthrene (3MC). Moreover, IPRN also increased estrogen receptor alpha (ERα) expression in an AhR-dependent manner. Taken together, these results suggest that IPRN acts as an AhR antagonist and promotes osteoblast differentiation via the AhR/ERα axis.

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Inhibitory Effects of Echinochrome A, Isolated from Shell of the Sea Urchin Anthocidaris crassispina, on Antigen-Stimulated Degranulation in Rat Basophilic Leukemia RBL-2H3 Cells through Suppression of Lyn Activation

Natural Product Communications ◽

10.1177/1934578x1601100930 ◽

2016 ◽

Vol 11 (9) ◽

pp. 1934578X1601100

Author(s):

Tomohiro Itoh ◽

Azusa Fujiwara ◽

Masayuki Ninomiya ◽

Toshimichi Maeda ◽

Masashi Ando ◽

...

Keyword(s):

Sea Urchin ◽

Intracellular Signaling ◽

Immunoglobulin E ◽

Inhibitory Effect ◽

Biologically Active ◽

Type I ◽

Dependent Manner ◽

Echinochrome A ◽

Basophilic Leukemia ◽

Anthocidaris Crassispina

Echinochrome A (Echi-A) was isolated from the sea urchin Anthocidaris crassispina and its structure determined using 1D and 2D-NMR. In the present study, we examined the inhibitory effect of Echi-A on antigen-stimulated degranulation in rat basophilic leukemia RBL-2H3 cells, which were suppressed in a dose dependent manner. The antigens bind to the high affinity immunoglobulin E receptor, which is expressed on the surface of mast cells and basophils and activate intracellular signal transduction, resulting in the release of biologically active mediators such as histamine. In order to disclose the inhibitory mechanisms of degranulation by Echi-A, we examined the elevation in intracellular Ca2+ concentration ([Ca2+]i), production levels of intracellular reactive oxygen species (ROS) and early intracellular signaling events. Both elevation of [Ca2+]i and intracellular ROS production were markedly suppressed in cells treated with Echi-A. Echi-A also suppressed the activation of Lyn, Syk, and PLCγ1/2 in antigen-stimulated cells. These results indicated that inhibition of antigen-stimulated degranulation in RBL-2H3 cells by Echi-A is mainly due to the inactivation of Lyn/Syk/PLCγ signaling pathways. Our findings suggest that Echi-A could be a beneficial agent for alleviating the symptoms of type I allergy.

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Phenazine-1-carboxylic acid, a secondary metabolite ofPseudomonas aeruginosa, alters expression of immunomodulatory proteins by human airway epithelial cells

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.00086.2003 ◽

2003 ◽

Vol 285 (3) ◽

pp. L584-L592 ◽

Cited By ~ 32

Author(s):

Gerene M. Denning ◽

Shankar S. Iyer ◽

Krzysztof J. Reszka ◽

Yunxia O'Malley ◽

George T. Rasmussen ◽

...

Keyword(s):

Carboxylic Acid ◽

Epithelial Cells ◽

Biological Effects ◽

Airway Epithelial Cells ◽

Biologically Active ◽

Bacterial Disease ◽

Airway Epithelial ◽

Disease Pathogenesis ◽

Human Airway ◽

Human Airway Epithelial Cells

Pseudomonas aeruginosa is a gram-negative bacterium that causes both acute and chronic lung disease in susceptible patient populations. P. aeruginosa secretes numerous proteins and secondary metabolites, many of which have biological effects that likely contribute to disease pathogenesis. An unidentified small-molecular-weight factor was previously reported to increase IL-8 release both in vitro and in vivo. To identify this factor, we subjected the <3-kDa fraction from P. aeruginosa-conditioned medium to HPLC analysis. A peak fraction that stimulated IL-8 release was found by mass spectrometry to have a molecular mass (MM) of 224 Da. On the basis of this MM and other biochemical properties, we hypothesized that the factor was phenazine-1-carboxylic acid (PCA). Subsequent studies and comparison with purified PCA confirmed this hypothesis. Purified PCA exhibited a number of biological effects in human airway epithelial cells, including increasing IL-8 release and ICAM-1 expression, as well as decreasing RANTES and monocyte chemoattractant protein-1 (MCP-1) release. PCA also increased intracellular oxidant formation as measured by electron paramagnetic resonance and by an intracellular oxidant-sensitive probe. Antioxidants inhibited PCA-dependent increases in IL-8 and ICAM-1, suggesting that oxidants contributed to these effects. However, in contrast to the related phenazine compound pyocyanin, PCA did not oxidize NAD(P)H at physiologically relevant pH, providing preliminary evidence that PCA and pyocyanin may have distinct redox chemistries within the cell. Thus PCA is a biologically active factor secreted by P. aeruginosa that has several activities that could alter the host immune and inflammatory response and thereby contribute to bacterial disease pathogenesis.

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Ovarian steroids affect prostaglandin production in equine endometrial cells in vitro

Journal of Endocrinology ◽

10.1530/joe-13-0185 ◽

2014 ◽

Vol 220 (3) ◽

pp. 263-276 ◽

Cited By ~ 12

Author(s):

Anna Z Szóstek ◽

António M Galvão ◽

Graça M Ferreira-Dias ◽

Dariusz J Skarzynski

Keyword(s):

Cell Proliferation ◽

Epithelial Cells ◽

Protein Expression ◽

Stromal Cells ◽

Positive Control ◽

Dependent Manner ◽

Ovarian Steroids ◽

Endometrial Cells ◽

Prostaglandin Endoperoxide Synthase

This study aimed to evaluate the influence of ovarian steroids on equine endometrial epithelial and stromal cells, specifically i) prostaglandin (PG) production in a time-dependent manner, ii) specific PG synthases mRNA transcription and protein expression, and iii) cell proliferation. After passage I, cells were exposed to vehicle, oxytocin (OT, positive control, 10−7M), progesterone (P4, 10−7M), 17β estradiol (E2, 10−9M), or P4+E2for 12, 24, 48, or 72 h. Following treatment, PG concentration was determined using the direct enzyme immunoassay (EIA) method. Alterations inPGsynthases mRNA transcriptions,PGsynthases protein expression, and cell proliferation in response to the treatments were determined after 24 h using real-time PCR, western blot, or 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide respectively. After 24 h, E2and P4+E2increased PGE2and PGF2αsecretion as well as specific prostaglandin-endoperoxide synthase-2 (PTGS2), PGE2synthases (PGES), and PGF2αsynthases (PGFS) expression in the epithelial cells (P<0.05). Additionally, E2and P4+E2increased PTGS2 expression in stromal cells after 24 h (P<0.05). In stromal cells, P4+E2increased PGE2production as well as PGES expression after 24 h (P<0.05). Both E2and P4+E2increased PGF2αproduction by stromal cells after 24 h (P<0.05). Ovarian steroids affected proliferation of stromal and epithelial cells during the 24-h incubation period (P<0.05). We provide evidence that ovarian steroids affect PG production in equine endometrial cells, upregulating PTGS2, PGES, and PGFS expression. Ovarian steroid-stimulated PG production could be an important mechanism occurring in the equine endometrium that is involved in the regulation of the estrous cycle and early pregnancy.

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Wnt5a signaling promotes apical and basolateral polarization of single epithelial cells

Molecular Biology of the Cell ◽

10.1091/mbc.e13-07-0357 ◽

2013 ◽

Vol 24 (23) ◽

pp. 3764-3774 ◽

Cited By ~ 17

Author(s):

Hidetoshi Gon ◽

Katsumi Fumoto ◽

Yonson Ku ◽

Shinji Matsumoto ◽

Akira Kikuchi

Keyword(s):

Epithelial Cells ◽

Cell Surface ◽

Molecular Mechanisms ◽

Type I Collagen ◽

Single Cells ◽

Surface Region ◽

Type I ◽

Dependent Manner ◽

Intestinal Epithelial ◽

Rac1 Activity

Single epithelial-derived tumor cells have been shown to induce apical and basolateral (AB) polarity by expression of polarization-related proteins. However, physiological cues and molecular mechanisms for AB polarization of single normal epithelial cells are unclear. When intestinal epithelial cells 6 (IEC6 cells) were seeded on basement membrane proteins (Matrigel), single cells formed an F-actin cap on the upper cell surface, where apical markers accumulated, and a basolateral marker was localized to the rest of the cell surface region, in a Wnt5a signaling–dependent manner. However, these phenotypes were not induced by type I collagen. Rac1 activity in the noncap region was higher than that in the cap region, whereas Rho activity increased toward the cap region. Wnt5a signaling activated and inhibited Rac1 and RhoA, respectively, independently through Tiam1 and p190RhoGAP-A, which formed a tertiary complex with Dishevelled. Furthermore, Wnt5a signaling through Rac1 and RhoA was required for cystogenesis of IEC6 cells. These results suggest that Wnt5a promotes the AB polarization of IEC6 cells through regulation of Rac and Rho activities in a manner dependent on adhesion to specific extracellular matrix proteins.

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Correlation between Phylogroups and Intracellular Proteomes ofPropionibacterium acnesand Differences in the Protein Expression Profiles between Anaerobically and Aerobically Grown Cells

BioMed Research International ◽

10.1155/2013/151797 ◽

2013 ◽

Vol 2013 ◽

pp. 1-9 ◽

Cited By ~ 11

Author(s):

Itaru Dekio ◽

Renata Culak ◽

Min Fang ◽

Graham Ball ◽

Saheer Gharbia ◽

...

Keyword(s):

Protein Expression ◽

Expression Profiles ◽

Opportunistic Pathogen ◽

Growth Conditions ◽

Culture Conditions ◽

Type I ◽

Proteomic Data ◽

Spectral Profiles ◽

Group B ◽

Protein Expression Profiles

Propionibacterium acnesis one of the dominant commensals on the human skin and also an opportunistic pathogen in relation to acne, sarcoidosis, prostate cancer, and various infections. Recent investigations using housekeeping and virulence genes have revealed that the species consists of three major evolutionary clades (types I, II, and III). In order to investigate protein expression differences between these phylogroups, proteomic profiles of 21 strains ofP. acneswere investigated. The proteins extracted from cells cultured under anaerobic and aerobic conditions were analysed using a SELDI-TOF mass spectrometer, high-resolution capillary gel electrophoresis, and LC-MS/ MS. The SELDI spectral profiles were visualised as a heat map and a dendrogram, which resulted in four proteomic groups. Strains belonging to type I were represented in the proteome Group A, while Group B contained type III strains. Groups C and D contained mixtures of types I and II. Each of these groups was not influenced by differences in culture conditions. Under anoxic growth conditions, a type IB strain yielded high expressions of some proteins, such as methylmalonyl-CoA epimerase and the Christie-Atkins-Munch-Petersen (CAMP) factor. The present study revealed good congruence between genomic and proteomic data suggesting that the microenvironment of each subtype may influence protein expression.

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