Journal: The Journal of toxicological sciences

Article Title: Involvement of P2Y receptors in the protective effect of ATP towards the cell damage in HaCaT cells exposed to H₂O₂.

doi: 10.2131/jts.36.741

Figure Lengend Snippet: Fig. 3. Effect of P2Y receptor inhibitors on ATP- and UTP- induced cytoprotection against H2O2-induced cell damage to HaCaT cells. Cells were incubated with ATP (A) or UTP (B) at a concentration of 10 μM for 6 hr in the presence or absence of MRS2179 (100 μM), MRS2578 (10 μM), NF157 (50 μM), clopidogrel (30 μM), MRS2211 (100 μM), or suramin (100 μM). After incubation, cells were exposed to H2O2 at a dose of 7.5 mM for 6 hr and cell damage was evaluated by means of LDH assay. The data represent the mean ± S.E. (n = 4). ### Indicates that experimental value is significantly different (p < 0.005) compared to the non- H2O2-exposed control group. *, **, and *** Indicate that experimental values are signifi cantly different (p < 0.05, p < 0.01, and p < 0.001, respectively) compared to the H2O2-alone-treated group. § Indicates that exper- imental value is signifi cantly different (p < 0.05) com- pared to the H2O2 plus ATP- or UTP-treated group.

Article Snippet: Suramin, A438079, MRS2179, MRS2578, NF157, clopidogrel hydrochloride and MRS2211 were from Tocris Bioscience (Bristol, UK).

Techniques: Incubation, Concentration Assay, Lactate Dehydrogenase Assay, Control

Journal: Molecular therapy : the journal of the American Society of Gene Therapy

Article Title: P2Y12 inhibitor clopidogrel inhibits renal fibrosis by blocking macrophage-to-myofibroblast transition.

doi: 10.1016/j.ymthe.2022.06.019

Figure Lengend Snippet: Figure 1. P2Y12 is expressed in the fibrotic kidney of patients with CKD and UUO mice (A) P2Y12 is significantly increased in patients with CKD with progressive interstitial fibrosis. (B) Correlation between eGFR and the expression of renal P2Y12 in 27 patients with CKD. (C) Western blot analysis shows that increased expression of renal P2Y12 on day 3 and 7 UUO kidney is associated with an increase in fibrosis markers of a-SMA and collagen I. (D) Real-time PCR shows expression of renal P2Y12 mRNA on day 3 and 7 kidney after UUO. Data represent mean ± SD from groups of 6 mice. (C and D) ***p < 0.001 versus sham, ##p < 0.01, ###p < 0.001 versus day 3 UUO mice. Scale bar, 25 mm.

Article Snippet: A P2Y12 inhibitor, clopidogrel bisulfate (Selleckchem S1415), was given to groups of 3 mice daily at dosages of 0, 20, 40, and 60 mg/kg orally from day 0 immediately after the ligation of left ureter to day 7.

Techniques: Expressing, Western Blot, Real-time Polymerase Chain Reaction

Journal: Molecular therapy : the journal of the American Society of Gene Therapy

Article Title: P2Y12 inhibitor clopidogrel inhibits renal fibrosis by blocking macrophage-to-myofibroblast transition.

doi: 10.1016/j.ymthe.2022.06.019

Figure Lengend Snippet: Figure 3. P2Y12 is expressed by MMT cells according to single-cell RNA sequencing data (A) Bubble plot shows the expression percentage (size) and relative expression levels (color) of cell lineage-specific markers and P2ry12 in each cell type. (B) Bar plot shows the ratio of P2ry12+ cells in each cell type. (C) Expression of selected maker genes in P2ry12+ cells.

Article Snippet: A P2Y12 inhibitor, clopidogrel bisulfate (Selleckchem S1415), was given to groups of 3 mice daily at dosages of 0, 20, 40, and 60 mg/kg orally from day 0 immediately after the ligation of left ureter to day 7.

Techniques: RNA Sequencing, Expressing

Journal: The Korean Journal of Pain

Article Title: Pharmacological interactions between intrathecal pregabalin plus tianeptine or clopidogrel in a rat model of neuropathic pain

doi: 10.3344/kjp.2022.35.1.59

Figure Lengend Snippet: Time course curves of intrathecal pregabalin (A), tianeptine (B) and clopidogrel (C) on the hind paw withdrawal response after spinal nerve ligation. Data are presented as the withdrawal threshold (g). Each line represents the means ± standard error of the mean of 6 rats.

Article Snippet: Co., Ltd., Seoul, Korea), and clopidogrel hydrogen sulfate (Tocris Bioscience, Bristol, UK); all drugs were dissolved in saline.

Techniques: Ligation

Journal: The Korean Journal of Pain

Article Title: Pharmacological interactions between intrathecal pregabalin plus tianeptine or clopidogrel in a rat model of neuropathic pain

doi: 10.3344/kjp.2022.35.1.59

Figure Lengend Snippet: Dose response effect of intrathecal pregabalin, tianeptine and clopidogrel (A), or mixture of pregabalin and tianeptine (B) on the hind paw withdrawal response after spinal nerve ligation. Data are presented as the percent of maximal possible effect (% MPE). Each line represents the means ± standard error of the mean of 6 rats. P: pregabalin ED 50 , T: tianeptine ED 50 , ED 50 : 50% effective dose. * P < 0.05, *** P < 0.001, compared with vehicle.

Article Snippet: Co., Ltd., Seoul, Korea), and clopidogrel hydrogen sulfate (Tocris Bioscience, Bristol, UK); all drugs were dissolved in saline.

Techniques: Ligation

Journal: The Korean Journal of Pain

Article Title: Pharmacological interactions between intrathecal pregabalin plus tianeptine or clopidogrel in a rat model of neuropathic pain

doi: 10.3344/kjp.2022.35.1.59

Figure Lengend Snippet: A fixed dose analysis for the interaction between intrathecal clopidogrel (300 μg) and pregabalin. Data are presented as the percent of maximal possible effect (% MPE). Each line represents the means ± standard error of the mean of 6 rats.

Article Snippet: Co., Ltd., Seoul, Korea), and clopidogrel hydrogen sulfate (Tocris Bioscience, Bristol, UK); all drugs were dissolved in saline.

Techniques:

Journal: bioRxiv

Article Title: Microglial P2Y12 mediates chronic stress-induced synapse loss in the prefrontal cortex and associated behavioral consequences in male mice

doi: 10.1101/2022.03.16.484487

Figure Lengend Snippet: A . Male wild-type or Thy1-GFP(M) mice were exposed to 14 days of chronic unpredictable stress (CUS) or were handled as controls. During this time, animals received daily injections of either vehicle, clopidogrel, or ticagrelor. One cohort was subjected to behavioral testing ( n = 9-20/group). B . Average time spent immobile in the forced swim test (FST). C . Average time spent exploring objects in the temporal object recognition task (TOR). D . Discrimination index in the TOR. E . In a separate cohort of mice, brains were extracted, frontal cortex was dissected out, and microglia were isolated and characterized using flow cytometry ( n = 7-22/group). Normalized mean fluorescence intensity of P2Y12, CSF1R, CD11b, and CX3CR1 in frontal cortex microglia. Normalized mean side scatter profile is shown. Bars represent mean ± S.E.M. * p <0.05 compared to same-treatment unstressed animal. # p <0.05 compared to unstressed vehicle-treated animal.

Article Snippet: Clopidogrel sulfate (LKT Laboratories, St Paul, MN; C4658) was diluted to 5 mg/ml in 0.9% sodium chloride.

Techniques: Isolation, Flow Cytometry, Fluorescence

Journal: bioRxiv

Article Title: Microglial P2Y12 mediates chronic stress-induced synapse loss in the prefrontal cortex and associated behavioral consequences in male mice

doi: 10.1101/2022.03.16.484487

Figure Lengend Snippet: Male Thy1-GFP(M) mice were exposed to 14 days of chronic unpredictable stress (CUS) or were handled as controls. During this time, animals received daily injections of either vehicle or clopidogrel. Approximately 4 hours after the final stressor, mice were perfused and brains were collected, sectioned, immunostained, and imaged ( n = 6-8/group). A . Confocal images of microglial IBA1 (red) and P2Y12 (cyan) in the mPFC. White scale bar represents 100 μm. B . Average number of FosB+ cells/mm 2 in the mPFC. C . Average number of IBA1+ cells/mm 2 in the mPFC. D . Average IBA1+ area per microglial cell. E . Average intensity (A.U.) of P2Y12 staining per microglial cell (relative to vehicle treated control group). F . Confocal images of microglia (IBA1, red) and dendritic segments (Thy1-GFP, green) were obtained from lamina I of the mPFC. Alongside merged channels, an orthogonal cross-section (matching the noted location) and representative dendritic segment is depicted for groups exposed to CUS. Microglial processes in close proximity to dendritic elements are noted within dashed circles, arrows indicate a dendritic element localized within a microglial cell body or process extension. White scale bar represents 5 μm. See Fig.S9 for additional images. G . Average dendritic spine density. H . Proportion of microglia with GFP+ inclusions. I . Number of GFP+ inclusions within microglia with dendritic elements. J . Average GFP+ inclusion volume per phagocytic microglia. * p <0.05 compared to same-treatment unstressed animal. # p <0.05 compared to unstressed vehicle-treated animal.

Article Snippet: Clopidogrel sulfate (LKT Laboratories, St Paul, MN; C4658) was diluted to 5 mg/ml in 0.9% sodium chloride.

Techniques: Staining, Control