Journal: Inflammatory bowel diseases

Article Title: Potential for Developing Purinergic Drugs for Gastrointestinal Diseases

doi: 10.1097/MIB.0000000000000047

Figure Lengend Snippet: Drugs in clinical trials targeting purinergic receptors

Article Snippet: N/A , STW5/II , A 2A AR Antag , IBS , Steigerwald Arzneimittelwerke GmbH , II , Completed 189 , Yes.

Techniques: Functional Assay, Inhibition

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Dexmedetomidine Attenuates Orthotopic Liver Transplantation-Induced Acute Gut Injury via α 2 -Adrenergic Receptor-Dependent Suppression of Oxidative Stress

doi: 10.1155/2019/9426368

Figure Lengend Snippet: Blocking α 2A -AR siRNA reversed the protective role of Dex on OLT-induced intestinal injury in rats. (a) Representative images showing the microscopic findings of intestines stained with hematoxylin and eosin in seven groups. Bar scale, 10 μ m. (b) Chiu's scoring quantitating the intestinal mucosal damage. (c–d) Immunohistochemistry staining of CD3 (c) and CD4 (d) in intestinal issues. Red arrow: positive cells. Data are represented as the mean ± standard deviation, n = 8 for each group. S: sham-operated group; M: the model group with OLT; D1: rats that were pretreated with 10 μ g/kg before OLT; D2: rats that were pretreated with 50 μ g/kg before OLT; B1: rats that received 500 g/kg atipamezole at 40 min before receiving 50 μ g/kg Dex prior to OLT; B2: rats that received 50 g/kg ARC239 at 40 min before receiving 50 μ g/kg Dex prior to OLT; B3: rats that received 1.5 mg/kg BRL-44408 at 40 min before receiving 50 μ g/kg Dex prior to OLT. ∗ P < 0.05, compared to Group S; # P < 0.05, compared to Group M; $ P < 0.05, compared to Group D2.

Article Snippet: The α 2A -AR siRNA duplexes (5′-CGAGCUGCAAGAUUAACGA-3′) targeting the rat α 2A -AR siRNA gene (gene ID: 25083; nucleotide accession number: NM_012739.3) were commercially obtained from Biomics (Jiangsu, China).

Techniques: Blocking Assay, Staining, Immunohistochemistry, Standard Deviation

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Dexmedetomidine Attenuates Orthotopic Liver Transplantation-Induced Acute Gut Injury via α 2 -Adrenergic Receptor-Dependent Suppression of Oxidative Stress

doi: 10.1155/2019/9426368

Figure Lengend Snippet: Blocking α 2A -AR siRNA reversed the protective role of Dex on OLT-induced intestinal mucosal barrier injury in rats. (a–b) Western blotting results showing the levels of occludin (a) and ZO-1 (b) in rat intestines. (c–f) ELISA results showing the levels of serum biomarkers of intestinal mucosal barrier function, including DAO (c), LPS (d), I-FABP2 (e), and D-LA (f). Data are represented as the mean ± standard deviation, n = 8 for each group. S: sham-operated group; M: the model group with OLT; D1: rats that were pretreated with 10 μ g/kg before OLT; D2: rats that were pretreated with 50 μ g/kg before OLT; B1: rats that received 500 g/kg atipamezole at 40 min before receiving 50 μ g/kg Dex prior to OLT; B2: rats that received 50 g/kg ARC239 at 40 min before receiving 50 μ g/kg Dex prior to OLT; B3: rats that received 1.5 mg/kg BRL-44408 at 40 min before receiving 50 μ g/kg Dex prior to OLT. ∗ P < 0.05, compared to Group S; # P < 0.05, compared to Group M; $ P < 0.05, compared to Group D2.

Article Snippet: The α 2A -AR siRNA duplexes (5′-CGAGCUGCAAGAUUAACGA-3′) targeting the rat α 2A -AR siRNA gene (gene ID: 25083; nucleotide accession number: NM_012739.3) were commercially obtained from Biomics (Jiangsu, China).

Techniques: Blocking Assay, Western Blot, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Dexmedetomidine Attenuates Orthotopic Liver Transplantation-Induced Acute Gut Injury via α 2 -Adrenergic Receptor-Dependent Suppression of Oxidative Stress

doi: 10.1155/2019/9426368

Figure Lengend Snippet: Blocking α 2A -AR siRNA counteracted the alleviation of oxidative stress by Dex in intestines of rats with OLT. The levels of oxidant ROS (a) and antioxidants including GST α 1 (b), SOD (c), and GSH (d) were measured in rat intestines. Data are represented as the mean ± standard deviation, n = 8 for each group. S: sham-operated group; M: the model group with OLT; D1: rats that were pretreated with 10 μ g/kg before OLT; D2: rats that were pretreated with 50 μ g/kg before OLT; B1: rats that received 500 g/kg atipamezole at 40 min before receiving 50 μ g/kg Dex prior to OLT; B2: rats that received 50 g/kg ARC239 at 40 min before receiving 50 μ g/kg Dex prior to OLT; B3: rats that received 1.5 mg/kg BRL-44408 at 40 min before receiving 50 μ g/kg Dex prior to OLT. ∗ P < 0.05, compared to Group S; # P < 0.05, compared to Group M; $ P < 0.05, compared to Group D2.

Article Snippet: The α 2A -AR siRNA duplexes (5′-CGAGCUGCAAGAUUAACGA-3′) targeting the rat α 2A -AR siRNA gene (gene ID: 25083; nucleotide accession number: NM_012739.3) were commercially obtained from Biomics (Jiangsu, China).

Techniques: Blocking Assay, Standard Deviation

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Dexmedetomidine Attenuates Orthotopic Liver Transplantation-Induced Acute Gut Injury via α 2 -Adrenergic Receptor-Dependent Suppression of Oxidative Stress

doi: 10.1155/2019/9426368

Figure Lengend Snippet: Silencing of α 2A -AR siRNA reversed the protective role of Dex on cell apoptosis in IEC-6 cells with simulated H/R stimulation. (a) qRT-PCR data showing effective knockdown of α 2A -AR by siRNA transfection in IEC-6 cells. Control: untreated IEC-6 cells; NC-siRNA: IEC-6 cells transfected with nonspecific control siRNA; α 2A -AR siRNA: IEC-6 cells transfected with α 2A -AR-specific siRNA. (b) Results of the cell proliferation assay by Cell Counting Kit-8 in IEC-6 cells with different stimulations. (c) Summary of cell apoptosis data of IEC-6 cells with different stimulations. (d) Representative flow cytometry profiles showing the cell apoptosis assay by annexin V and propidium iodide staining, n = 6 for each group. A: control IEC-6 cells; B: IEC-6 cells with H/R treatment; C: IEC-6 cells that were pretreated with 1 nM Dex for 1 h before inducing H/R injury; D: IEC-6 cells with silencing of α 2A -AR siRNA; E: IEC-6 cells with silencing of α 2A -AR siRNA and H/R treatment; F: IEC-6 cells with silencing of α 2A -AR siRNA that were pretreated with Dex prior to H/R injury. ∗ P < 0.05, compared to Group A; # P < 0.05, compared to Group B; $ P < 0.05, compared to Group C.

Article Snippet: The α 2A -AR siRNA duplexes (5′-CGAGCUGCAAGAUUAACGA-3′) targeting the rat α 2A -AR siRNA gene (gene ID: 25083; nucleotide accession number: NM_012739.3) were commercially obtained from Biomics (Jiangsu, China).

Techniques: Quantitative RT-PCR, Knockdown, Transfection, Control, Proliferation Assay, Cell Counting, Flow Cytometry, Apoptosis Assay, Staining

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Dexmedetomidine Attenuates Orthotopic Liver Transplantation-Induced Acute Gut Injury via α 2 -Adrenergic Receptor-Dependent Suppression of Oxidative Stress

doi: 10.1155/2019/9426368

Figure Lengend Snippet: Silencing of α 2A -AR siRNA erased the protective role of Dex on antioxidation in IEC-6 cells with simulated H/R stimulation. (a) Summary of ROS levels in IEC-6 cells with different treatments. (b) Representative fluorescence immunostaining images of Nrf2 in IEC-6 cells with different treatments. Scale bar, 100 μ m. (c–d) Expression of Nrf2, NQO1, and Keap1 proteins and mRNA levels in intestinal tissues. (e–g) Summary of the relative expression levels of the transcripts for the genes HMOX1 (e), SOD1 (f), and CAT (g) in IEC-6 cells with different treatments, n = 6 for each group. A: control IEC-6 cells; B: IEC-6 cells with H/R treatment; C: IEC-6 cells that were pretreated with 1 nM Dex for 1 h before inducing H/R injury; D: IEC-6 cells with silencing of α 2A -AR siRNA; E: IEC-6 cells with silencing of α 2A -AR siRNA and H/R treatment; F: IEC-6 cells with silencing of α 2A -AR siRNA that were pretreated with Dex prior to H/R injury. ∗ P < 0.05, compared to Group A; # P < 0.05, compared to Group B; $ P < 0.05, compared to Group C.

Article Snippet: The α 2A -AR siRNA duplexes (5′-CGAGCUGCAAGAUUAACGA-3′) targeting the rat α 2A -AR siRNA gene (gene ID: 25083; nucleotide accession number: NM_012739.3) were commercially obtained from Biomics (Jiangsu, China).

Techniques: Fluorescence, Immunostaining, Expressing, Control

Journal: Nature Communications

Article Title: Specific pharmacological and G i/o protein responses of some native GPCRs in neurons

doi: 10.1038/s41467-024-46177-z

Figure Lengend Snippet: a Effect of the indicated α 2 AR agonists on the net BRET of the G i1 and G oA sensors in CGNs co-transfected with Gα i1 Nluc and Venus Gγ 2 or Gα oA Nluc and Venus Gγ 2 (amounts of cDNA as in Fig. ). b Effect of the indicated α 2 AR agonists on the net BRET of the G i1 and G oA sensors in HEK293 cells co-transfected with the mouse α 2A AR (10 ng, 20 ng or 50 ng), and Gβ 1, Venus Gγ 2 and Gα i1 Nluc or Gα oA Nluc as in Fig. . Saturating concentrations of brimonidine (10 μM), oxymetazoline (100 μM), xylazine (50 μM), clonidine (100 μM), tizanidine (100 μM) were used in ( a , b ). c Detection of α 2 AR in cell membranes of CGNs, mock-transfected HEK293 cells and HEK293 cells transfected with indicated amount of α 2 AR, by western blotting. Values are mean ± SEM normalized as fold of α 2 AR expression in CGNs from four independent experiments. d The pEC 50 of brimonidine in CGNs ( n = 3) and transfected HEK293 cells ( n = 4) with indicated amount of α 2 AR measured by G i1 or G oA sensor. e Percentage of change in BRET ratio between Gα Nluc and Venus Gγ 2 induced by brimonidine between Gα Nluc and Venus Gγ 2 in HEK293 cells ( n = 3) (α 2 AR: 15 ng/well per 96-well plate) or CGNs ( n = 4) for the indicated Gα i1 , Gα i2 , Gα i3 , Gα oA , Gα oB or Gα z sensors (amounts of cDNA as in Fig. ). Values are mean ± SEM from at least three biologically independent experiments each performed in triplicate or quadruplicate in ( a , b , d , e ). a , n = 4; b , n = 3; ( d , e ), CGNs, n = 3; HEK293 cells, n = 4. Data are analysed using one-way ANOVA with a Dunnett’s post-hoc multiple comparison test to determine significance (compared with brimonidine in a , b ). Data are analysed using unpaired t -test (two-tailed) in ( e ). **** p < 0.0001, *** p < 0.001, ** p < 0.01 and not significant (ns). The raw data and p -values are available in source data provided as a Source Data file.

Article Snippet: The blots were incubated with the primary monoclonal antibodies anti-GB1 mAb (1:1000, ab55051, Abcam, Shanghai, China), anti-Gβ 2 rabbit (1:1000, A9643, ABclonal Technology, Wuhan, China), anti-β-actin (1:3000, KM9001T, Sungene Biotech., Tianjin, China), the rabbit polyclonal antibodies anti-CB1 (1:1000, A1447, ABclonal, Wuhan, China), anti-α 2A AR (1:1000, A2809, ABclonal, Wuhan, China), anti-Gβ 1 (1:1000, A1867, ABclonal, Wuhan, China), anti-Gβ 3 (1:1000, A1387, ABclonal, Wuhan, China) and anti-Gβ 5 (1:1000, A4447, ABclonal, Wuhan, China).

Techniques: Transfection, Western Blot, Expressing, Comparison, Two Tailed Test