Journal: Heliyon

Article Title: Role of plasmacytoid dendritic cells in vascular dysfunction in mice with renovascular hypertension

doi: 10.1016/j.heliyon.2024.e31799

Figure Lengend Snippet: Mesenteric resistance arteries (MRA) Reactivity Assessment. This figure delineates the evaluation of mesenteric resistance arteries (MRA) reactivity, specifically illustrating the contractility response to sympathetic stimulation (Phenylephrine, PE), as well as endothelium-dependent and -independent relaxation responses to acetylcholine (ACh) and sodium nitroprusside (SNP) across diverse experimental groups. Panel (A, B) represents the control group (C57BL/6J), while panel (C, D) shows C57BL/6J mice subjected to 2K1C surgery. Panel (E, F) displays C57BL/6J mice subjected to 2K1C for four weeks and treated with anti-PDCA-1 for one week. MRA reactivity was thoroughly evaluated in all groups of mice (n = 5) under different conditions, including those with and without ER stress inhibitor (Tauroursodeoxycholic acid: Tudca), autophagy inhibitor (Chloroquine: Chl), and mTOR signaling inhibitor (Rapamycin: Rap). The data presented in this figure provide valuable insights into the vascular response in the context of the experimental manipulations and treatments, shedding light on the potential involvement of ER stress, autophagy, and mTOR signaling pathways in mediating the observed reactivity changes. ns: p > 0.05, *p < 0.05 for 2K1C vs. control and 2K1C + anti-mPDCA-1. One-way ANOVA followed by Tukey's post hoc test was applied. ns: p > 0.05, *p < 0.05 for C57BL/6J vs 2K1C vs 2K1C + anti-PDCA-1.

Article Snippet: To determine the impact of autophagy, ER stress, and mTOR pathways in endothelial cell function, we isolated arteries from each group and incubated them with the following inhibitors: tauroursodeoxycholic acid (Tudca: ER Stress inhibitor) Dose: 10 mM, 30 min, Chem-Impex Int'l INC. Cat#29195, Lot# 002129–20131118; chloroquine (autophagy inhibitor, dose: 10 mM, 30 min, Alfa Aesar, Cat: J64459, Lot: Z23G009); or rapamycin (mTOR inhibitor, dose: 10 mM, 30 min, Abcam, Cat:ab120224, Lot: APN13087-1-1).

Techniques: Control

Journal: Molecules

Article Title: Cyclometalated Iridium(III) Complex–Cationic Peptide Hybrids Trigger Paraptosis in Cancer Cells via an Intracellular Ca 2+ Overload from the Endoplasmic Reticulum and a Decrease in Mitochondrial Membrane Potential

doi: 10.3390/molecules26227028

Figure Lengend Snippet: ( A ) The results (cell survival) of MTT assays of Jurkat cells treated with ASb-2 (50 µM) in the presence of CCCP (40 µM), rapamycin (200 nM), cycloheximide (2 µM), 2-APB (50 µM), BAPTA-AM (10 µM), and TUDCA (40 µM) for 1 h at 37 °C. ( B ) Typical microscopy images (Biorevo, BZ-9000, Keyence) of Jurkat cells treated with ASb-2 (50 µM) in the presence of CCCP (40 µM), rapamycin (200 nM), cycloheximide (2 µM), 2-APB (50 µM), BAPTA-AM (10 µM) and TUDCA (40 µM) for 1 h at 37 °C. Scale bar: 20 µm ( C ) The results of MTT assays of Jurkat cells treated with celastrol in the presence of CCCP (40 µM), rapamycin (200 nM), cycloheximide (2 µM), 2-APB (50 µM), BAPTA-AM (10 µM) and TUDCA (40 µM) for 24 h at 37 °C. * p < 0.05, ** p < 0.01, and **** p < 0.0001 indicate significant differences from the control. ns: no significant (the statistical data in A,C are listed in ).

Article Snippet: Celastrol, U0126, 2-aminoethoxydiphenyl borate (2-APB), SP600125, SCH772984, SB203580, and tauroursodeoxycholic acid (TUDCA) were purchased from Cayman Chemical Co. (Ann Arbor, MI, USA).

Techniques: Microscopy

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: The Food Additive Maltodextrin Promotes Endoplasmic Reticulum Stress–Driven Mucus Depletion and Exacerbates Intestinal Inflammation

doi: 10.1016/j.jcmgh.2018.09.002

Figure Lengend Snippet: ER stress inhibition reduces MDX-mediated Ern-2 up-regulation and improves colitis in MDX-fed mice. ( A ) HT29-MTX cells were pretreated with TUDCA (10 μmol/L) or dimethyl sulfoxide (vehicle) and then stimulated with MDX for 1 hour. Ern-2 RNA transcripts were analyzed by real-time PCR. Data are means ± SD of 4 independent experiments. Differences between groups were compared using the 2-tailed Student t test (* P ≤ .05). ( B and C ) Wild-type mice were exposed to drinking water supplemented with 5% MDX for 45 days and injected or not with TUDCA (250 mg/kg intraperitoneally) every other day starting from day 21. Mice were killed on day 45, colonic tissues were isolated, and ( B ) Ern-2 , Ern-1 , and Xbp1s RNA transcripts and ( C ) Muc-2 protein expression were evaluated by real-time PCR and immunofluorescence, respectively. ( B ) Data were generated using 7–10 mice per group from 3 independent experiments. Each point in the graph indicates the RNA expression of the specific transcript in the colon of a single mouse; horizontal bars indicate median value. Differences between groups were compared using the Mann–Whitney U test (* P ≤ .05). ( C ) Pictures are representative of 4 separate experiments in which similar results were obtained. Scale bars : 25 μm. ( D ) Wild-type mice were exposed to drinking water supplemented with 5% MDX for 45 days and injected or not with TUDCA (250 mg/kg intraperitoneally) every other day starting from day 21. Mice were exposed to 1.75% DSS to induce colitis starting from day 35 until death (day 45), and body weight was recorded every other day. Data were generated using 8–9 mice per group from 3 independent experiments and expressed as means ± SEM. Differences among groups were compared using 1-way analysis of variance followed by the Bonferroni post hoc test (** P ≤ .01). ( E and F ) Representative H&E staining of colon sections of mice treated as indicated in panel D and killed on day 45. ( F ) Scatter plot shows the histologic score. Data were generated using 8–9 mice per group from 3 independent experiments and expressed as means ± SD. Differences among groups were compared using 1-way analysis of variance followed by the Bonferroni post hoc test (* P ≤ .05; ** P ≤ .01). ( G and H ) Representative scatter plots showing ( G ) IL1β and ( H ) Lcn-2 RNA expression in colon tissues taken from mice treated as indicated in panel D and killed on day 45. Each point in the graph indicates the RNA expression of the specific transcript in the colon of a single mouse; horizontal bars indicate median value or means ± SD. Data were generated using 8–9 mice per group from 3 independent experiments. Differences among groups were compared using the Kruskal–Wallis test or 1-way analysis of variance followed by the Bonferroni post hoc test (* P ≤ .05). DAPI, 4′,6-diamidino-2-phenylindole; mRNA, messenger RNA.

Article Snippet: In parallel, mice receiving a MDX-enriched diet, together with control mice, were given 250 mg/kg TUDCA (Carbosynth Ltd, Berkshire, UK) intraperitoneally every other day starting from day 21 of diet.

Techniques: Inhibition, Real-time Polymerase Chain Reaction, Injection, Isolation, Expressing, Immunofluorescence, Generated, RNA Expression, MANN-WHITNEY, Staining

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: The Food Additive Maltodextrin Promotes Endoplasmic Reticulum Stress–Driven Mucus Depletion and Exacerbates Intestinal Inflammation

doi: 10.1016/j.jcmgh.2018.09.002

Figure Lengend Snippet: Effect of MDX on mucosa-associated microbiota. ( A and B ) Wild-type mice were exposed to drinking water supplemented with 5% MDX for 45 days and injected or not with TUDCA (250 mg/kg intraperitoneally) every other day starting from day 21. Control mice received drinking water for 45 days. All mice were killed on day 45. ( A ) Relative abundance of phyla and classes are represented for colonic mucosa-associated microbiota. Horizontal bars indicate median value. Data were generated using 4–7 mice per group from 2 independent experiments. Differences among groups were compared using the Kruskal–Wallis test (* P ≤ .05). ( B ) Principal coordinates analysis (PCoA) of the unweighted and weighted UniFrac distance matrix of mucosa-associated bacteria from mice treated as indicated in panel A .

Article Snippet: In parallel, mice receiving a MDX-enriched diet, together with control mice, were given 250 mg/kg TUDCA (Carbosynth Ltd, Berkshire, UK) intraperitoneally every other day starting from day 21 of diet.

Techniques: Injection, Control, Generated, Bacteria

Journal: Microbiome

Article Title: Antibiotic-modulated microbiome suppresses lethal inflammation and prolongs lifespan in Treg-deficient mice

doi: 10.1186/s40168-019-0751-1

Figure Lengend Snippet: Antibiotics regulate cytokines by microbiota-associated metabolites. a – d Plasma levels of IFN-γ ( a ), IL-2( b ), IL-6( c ), and TNF- α( d ) in WT, SF, SFA, SFM, and SFV mice ( n = 6–9). e Level of IL-6 in RAW264.7 cells treated with DMSO (blank), LPS, taurocholic acid sodium salt hydrate (TCA), sodium tauroursodeoxycholate (TUDCA), and taurochenodeoxycholic acid (TCDCA) ( n = 3). Data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: For determining the effect of bile acids on IL-6 expression and cell viability in RAW 264.7 murine macrophage cells, after 24 h from splitting 3000 cells into one well of 96-well plates, cells were pretreated with taurocholic acid sodium salt hydrate (Sigma), sodium tauroursodeoxycholate (Selleck), and taurochenodeoxycholic acid (Selleck) (5, 25, and 125 μM) for 2 h. Subsequently, the cells were stimulated with 50 ng/mL lipopolysaccharide (LPS) for 12 h. Following this, the concentration of IL-6 in the supernatant was measured by IL-6 mouse ELISA kit (Thermo Fisher) and cell viability was measured by TACS XTT cell proliferation assay kit (Trevigen, Inc.).

Techniques: Clinical Proteomics

Journal: Microorganisms

Article Title: An Analysis of the Gut Microbiota and Related Metabolites following PCSK9 Inhibition in Statin-Treated Patients with Elevated Levels of Lipoprotein(a)

doi: 10.3390/microorganisms12010170

Figure Lengend Snippet: ( A – C ) Boxplots representing the concentration (log10 transformed) of total bile acids ( A ), and primary ( B ) and secondary ( C ) bile acids in fecal samples collected at baseline and on treatment. Samples from the same patient are connected using straight lines. The following primary bile acids were detected in feces: cholic acid, taurocholic acid, chenodeoxycholic acid, taurochenodeoxycholic acid and glycochenodeoxycholic acid. The following secondary bile acids were detected in feces: deoxycholic acid, lithocholic acid, ursodeoxycholic acid, taurolithocholic acid, tauroursodeoxycholic acid, taurodeoxycholic acid and glycodeoxycholic acid. For statistical inference, we used a paired t -test, and the corresponding p -values are shown in each graph. ( D ) Linear relationship between the reduction in plasma LDL-c and the concentration of fecal bile acids (log10 transformed) after alirocumab therapy. Line of best fit for the linear model is plotted with 95% confidence interval (grey shade area). Individual samples are represented with black dots. The regression coefficient, coefficient of determination and the result of the F-test for the linear model are shown on the top of the plot.

Article Snippet: For identification of the different classes of BAs, we used the following pure standards to create a mass spectral–retention time library; taurocholic acid (Santa Cruz, sc220189, Dallas, TX, USA), taurolithocholic acid (Cayman Chemicals, 17275, Ann Arbor, MI, USA), taurochenodeoxycholic acid (Steraloids, C1162-000, Newport, RI, USA), tauroursodeoxycholic acid (Steraloids, C1052-000), glycolithocholic acid (Steraloids, C1435-000), glycochenodeoxycholic acid (Steraloids, C0962-000), glycoursodeoxycholic acid (Steraloids, C1025-000), glycocholic acid (Steraloids, C1927-000), glycodeoxycholic acid (Steraloids, C1087-000), ursodeoxycholic acid (Steraloids, C1020-000), cholic acid (Sigma-Aldrich, C1129, St. Louis, MO, USA), deoxycholic acid (Sigma-Aldrich, D2510), lithocholic acid (Sigma-Aldrich, L6250), chenodeoxycholic acid (Sigma-Aldrich, C1050000).

Techniques: Concentration Assay, Transformation Assay, Clinical Proteomics

Journal: Human Molecular Genetics

Article Title: TFIIH mutations can impact on translational fidelity of the ribosome

doi: 10.1093/hmg/ddac268

Figure Lengend Snippet: Rescue experiments with N-acetyl-cysteine and the pharmaceutical chaperone TUDCA. TUDCA restores RNA polymerase I transcription, rRNA processing and protein synthesis in TTD cells. ( A ) Plasmids encoding for renilla and firefly luciferase were transcribed to capped mRNAs by T7 polymerase. Reporter mRNAs were transfected (lipofectamine) in the transformed TTD cell lines P8 Mut , P8 Rec , XPD Mut and XPD Rec pretreated with 1 mM N-acetyl-cysteine for 24 h. ( B ) 24 h pre-treatment of p8- and XPD-mutated cells with TUDCA (200 μM) followed by quantitative PCR analysis of 47S pre-rRNA expression. ( C ) Protein synthesis analysis after TUDCA treatment. ( D ) Reduction of processing defects measured by ITS1 accumulation by 24 h treatment with TUDCA (200 μM). Quantitative PCR analysis of the ITS1 region. Values are represented as mean ± SD of at least three independent experiments. Asterisk ( * ) in the figure represent ( * P < 0.05, * * P < 0.01, * * * P < 0.001, * * * * P < 0.0001)

Article Snippet: For chemical chaperones, cells were treated for 24 h with TUDCA (200 μM) (Tauroursodeoxycholic Acid, Sodium Salt Affiliate of Merck Darmstadt, Germany) 4PBA (1 mM) (4-Phenylbutyric acid Sigma-Aldrich, P21005) and were cultured under standard conditions at 37°C, 5%; CO 2 and 3%; oxygen in DMEM media.

Techniques: Luciferase, Transfection, Transformation Assay, Real-time Polymerase Chain Reaction, Expressing

Journal: PLoS ONE

Article Title: Increased Susceptibility to Dextran Sulfate Sodium-Induced Colitis in the Endoplasmic Reticulum Stress Transducer OASIS Deficient Mice

doi: 10.1371/journal.pone.0088048

Figure Lengend Snippet: All the Oasis −/− mice received 3.5% DSS and some were given TUDCA (+TUDCA) and others were given the same volume of PBS (vehicle) daily by oral administration for 5 days. (A) HE (upper panels) and PAS (lower panels) staining of the large intestinal mucosa of Oasis −/− mice exposed to 3.5% DSS and TUDCA or the vehicle. (B) Higher magnification of HE staining in (A). Arrowheads show inflammatory cells. (C) Histological scores of control and TUDCA-treated Oasis −/− mice that received 3.5% DSS ( n = 4). The pathological findings were markedly improved in Oasis −/− mice treated with TUDCA. (D) RT-PCR analysis of Bip and Chop in the large intestinal mucosa of Oasis −/− mice. The expression levels of these ER stress markers in the large intestinal mucosa of Oasis −/− mice were decreased by treatment with TUDCA. (E) Quantification of the expression levels of Bip and Chop in (D) ( n = 4). (F) RT-PCR analysis of inflammatory cytokines in the large intestinal mucosa of Oasis −/− mice exposed to 3.5% DSS and TUDCA or the vehicle for 5 days ( n = 4). Note that the expression levels of inflammatory cytokines were decreased in Oasis −/− mice that received TUDCA. (G) Western blotting of cleaved caspase-12 and -3 in Oasis −/− large intestinal mucosa exposed to 3.5% DSS and TUDCA or the vehicle for 5 days. (H) TUNEL staining of the large intestinal mucosa in Oasis −/− mice that received 3.5% DSS and TUDCA or the vehicle. (I) The number of TUNEL-positive cells in (H) (n = 4). The number of TUNEL-positive cells was decreased in Oasis −/− mice treated with TUDCA. Values represent the means ± s.d. * P <0.05; ** P <0.01.

Article Snippet: Tauroursodeoxycholic acid (TUDCA) (10 mg/ml, sodium salt; Wako) in sterilized PBS was administered orally (160 mg/kg/day) for 5 days.

Techniques: Staining, Reverse Transcription Polymerase Chain Reaction, Expressing, Western Blot, TUNEL Assay

Journal: PLoS ONE

Article Title: Increased Susceptibility to Dextran Sulfate Sodium-Induced Colitis in the Endoplasmic Reticulum Stress Transducer OASIS Deficient Mice

doi: 10.1371/journal.pone.0088048

Figure Lengend Snippet: (A and B) RT-PCR analysis of (A) Bip and (B) Chop in LS174T human colon carcinoma cells treated with ER stress inducer tunicamycin (Tm) and TUDCA for 24 h ( n = 4). The expression levels of both Bip and Chop were downregulated by TUDCA. (C–E) RT-PCR analysis of (C) Tnfα , (D) IL-1 , and (E) IL-6 in LS174T cells treated with Tm and TUDCA ( n = 4). Note that the expression levels of these inflammatory cytokines were upregulated in LS174T cells treated with Tm, and downregulated by treatment with TUDCA. (F) Luciferase assay using LS174T cells transfected with the p-Luc reporter plasmid containing the NF-κB binding sequence. Relative activities were increased by treatment of LS174T cells with Tm, and decreased by treatment with TUDCA ( n = 4). Values represent the means ± s.d. * P <0.05; **P<0.01; *** P <0.001.

Article Snippet: Tauroursodeoxycholic acid (TUDCA) (10 mg/ml, sodium salt; Wako) in sterilized PBS was administered orally (160 mg/kg/day) for 5 days.

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Luciferase, Transfection, Plasmid Preparation, Binding Assay, Sequencing