Journal: Nature Communications

Article Title: Neutrophil P2X 7 receptors mediate NLRP3 inflammasome-dependent IL-1β secretion in response to ATP

doi: 10.1038/ncomms10555

Figure Lengend Snippet: ( a ). Western blot analysis of P2X 7 receptors on unstimulated and LPS-primed bone marrow neutrophils and macrophages from C57BL/6 and P2X 7 −/− mice. ( b – e ). Flow cytometry and fluorescence microscopy of P2X 7 R ecto domain (using the Hano-3 antibody) in LPS-primed bone marrow neutrophils that were either permeabilized with 0.1% TX-100 to detect total P2X 7 R ( b , c ) or were non-permeabilized to detect only cell surface P2X 7 R ( d , e ). ( c,e ) Cells were also stained with DAPI to detect the nucleus; Scale bar, 5 μm). ( f – h ). Representative profiles of ATP-induced Ca 2+ influx in bone marrow neutrophils from C57BL/6 and P2X 7 −/− mice ( f ), and in C57BL/6 neutrophils after stimulation with ATP (3 mM) in the presence of P2X 7 R antagonists AZ10606120 (10 μM) ( g ), or with different concentrations of A438079 ( h ). ( i – k ). Mean±s.e.m. pf Area under the curve (AUC) of intracellular Ca 2+ from three independent experiments. A P value ≤0.05 was considered significant using an unpaired Student's t- test.

Article Snippet: P2X 7 R inhibitors AZ10606120 dihydrochloride and AZ11645373, A438079 hydrochloride (Tocris Bioscience) were dissolved in DMSO and used at concentrations noted in the results.

Techniques: Western Blot, Flow Cytometry, Fluorescence, Microscopy, Staining

Journal: Nature Communications

Article Title: Neutrophil P2X 7 receptors mediate NLRP3 inflammasome-dependent IL-1β secretion in response to ATP

doi: 10.1038/ncomms10555

Figure Lengend Snippet: Total IL-1β secreted by bone marrow neutrophils from C57BL/6 and P2X 7 −/− mice ( a , b ), or from C57BL/6 neutrophils incubated with the P2X 7 antagonists AZ10606120 (10 μM) ( c ) or A438079 (25 μM) ( d ) following LPS priming and stimulation with ATP or nigericin measured by ELISA. ( e – g ). Bioactive IL-1 from the same samples detected using HEK-Blue-IL-1R reporter cells. ( h ). IL-1α and IL-1β from the C57BL/6 neutrophil supernatants stimulated 45 min with 3 mM ATP and 10 μM nigericin were quantified by ELISA. Data points are mean±s.d. of at least three replicates per treatment and are representative of three independent experiments. Using an ANOVA with Tukey post hoc analysis, a P value ≤0.05 was considered significant.

Article Snippet: P2X 7 R inhibitors AZ10606120 dihydrochloride and AZ11645373, A438079 hydrochloride (Tocris Bioscience) were dissolved in DMSO and used at concentrations noted in the results.

Techniques: Incubation, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Molecular Neuroscience

Article Title: The P2X 7 Receptor Is Involved in Diabetic Neuropathic Pain Hypersensitivity Mediated by TRPV1 in the Rat Dorsal Root Ganglion

doi: 10.3389/fnmol.2021.663649

Figure Lengend Snippet: The effect of A438079 on the mechanical withdrawal threshold (MWT) and the thermal withdrawal latency (TWL) of DNP rats. (A) Changes in the MWTs of rats from each group. (B) Changes in the TWLs of rats from each group. The MWTs and TWLs of rats from the DNP group were significantly lower than the values of rats from the control group (namely, the sensitivity was increased). At the end of the 8th week, the MWTs and TWLs of rats in the DNP group treated with an intrathecal injection of the P2X 7 receptor antagonist A438079 (DNP + A438079 group) were significantly higher than those of rats in the DNP group (i.e., decreased sensitivity). The data are presented as the mean ± SEM for the six animals in each group; compared with the control group, ** P < 0.01; compared with the DNP group, # P < 0.05, ## P < 0.01.

Article Snippet: In this experiment, the rats were divided into a control group (control group), DNP model group (DNP group), P2X 7 receptor antagonist A438079 (S7705; Selleckchem)-treated DNP group (DNP + A438079 group), and normal saline (NS)-treated DNP group (DNP + NS group).

Techniques: Control, Injection

Journal: Frontiers in Molecular Neuroscience

Article Title: The P2X 7 Receptor Is Involved in Diabetic Neuropathic Pain Hypersensitivity Mediated by TRPV1 in the Rat Dorsal Root Ganglion

doi: 10.3389/fnmol.2021.663649

Figure Lengend Snippet: The levels of the TRPV1 mRNA and protein in the DRG were detected. (A) The level of the TRPV1 mRNA in the DRG from each group. (B) SDS-PAGE band of the TRPV1 protein in the DRG from each group of experimental rats. (C) Analysis of the relative level of the TRPV1 protein in the DRG from each group of experimental rats. Higher levels of the TRPV1 mRNA and protein were detected in the DRG in the DNP group than in the control group, while lower levels of the TRPV1 mRNA and protein were observed in the DRG in the DNP + A438079 group than in the DNP group. The data are presented as the mean ± SEM, n = 6; compared with the control group, ** P < 0.01; compared with the DNP group, ## P < 0.01.

Article Snippet: In this experiment, the rats were divided into a control group (control group), DNP model group (DNP group), P2X 7 receptor antagonist A438079 (S7705; Selleckchem)-treated DNP group (DNP + A438079 group), and normal saline (NS)-treated DNP group (DNP + NS group).

Techniques: SDS Page, Control

Journal: Frontiers in Molecular Neuroscience

Article Title: The P2X 7 Receptor Is Involved in Diabetic Neuropathic Pain Hypersensitivity Mediated by TRPV1 in the Rat Dorsal Root Ganglion

doi: 10.3389/fnmol.2021.663649

Figure Lengend Snippet: The coexpression of TRPV1 with NeuN and P2X 7 receptor with GFAP was detected by immunofluorescence staining. (A) Levels of TRPV1 with NeuN coexpression in neurons in the rat DRG ( n = 6 rats for each group). (B) The histogram shows the mean optical density of the coexpression of TRPV1 with NeuN. (C) Levels of P2X 7 with GFAP coexpression in SGCs in the rat DRG ( n = 6 rats for each group). (D) The histogram shows the mean optical density of the coexpression of the P2X 7 receptor with GFAP. In the DNP group, the coexpression of TRPV1 with NeuN in the DRG was increased compared with that in the control group, while the coexpression of TRPV1 with NeuN in the DNP + A438079 group was decreased compared with that in the DNP group. Furthermore, the coexpression of P2X 7 receptor with GFAP in the DRG was consistent with the coexpression trend described above. The green signal is fluorescein isothiocyanate (FITC)-labeled NeuN and FITC-labeled GFAP; the red signal is tetramethylrhodamine isothiocyanate (TRITC)-labeled TRPV1 and TRITC-labeled P2X 7 ; and the yellow signal is the combination of the green and red signals. The arrow indicates activated neuronal cells and SGCs in the DRG. The scale bar represents 50 μm. The data are presented as the mean ± SEM; compared with the control group, ** P < 0.01; compared with the DNP group, ## P < 0.01.

Article Snippet: In this experiment, the rats were divided into a control group (control group), DNP model group (DNP group), P2X 7 receptor antagonist A438079 (S7705; Selleckchem)-treated DNP group (DNP + A438079 group), and normal saline (NS)-treated DNP group (DNP + NS group).

Techniques: Immunofluorescence, Staining, Control, Labeling

Journal: Frontiers in Molecular Neuroscience

Article Title: The P2X 7 Receptor Is Involved in Diabetic Neuropathic Pain Hypersensitivity Mediated by TRPV1 in the Rat Dorsal Root Ganglion

doi: 10.3389/fnmol.2021.663649

Figure Lengend Snippet: The relative levels of IL-1β and IL-10 in the DRG were detected by Western blotting. (A) SDS-PAGE band of IL-1β in the DRG in each group of experimental rats. (B) The relative levels of IL-1β in the DRG from each group of experimental rats. (C) SDS-PAGE band of IL-10 in the DRG in each group of experimental rats. (D) The relative levels of IL-10 protein in the DRG from each group of experimental rats. The relative level of the IL-1β protein in the DRG from the DNP group was higher than that in the control group, while a lower level of IL-1β was observed in the DRG from the DNP + A438079 group than in the DNP group. In contrast, the relative level of the IL-10 protein in the DRG from the DNP group was lower than that in the control group, while a higher level of IL-10 was detected in the DRG in the DNP + A438079 group than in the DNP group. The data are presented as the mean ± SEM, n = 6; compared with the control group, * P < 0.05, ** P < 0.01; compared with the DNP group, # P < 0.05, ## P < 0.01.

Article Snippet: In this experiment, the rats were divided into a control group (control group), DNP model group (DNP group), P2X 7 receptor antagonist A438079 (S7705; Selleckchem)-treated DNP group (DNP + A438079 group), and normal saline (NS)-treated DNP group (DNP + NS group).

Techniques: Western Blot, SDS Page, Control

Journal: Frontiers in Molecular Neuroscience

Article Title: The P2X 7 Receptor Is Involved in Diabetic Neuropathic Pain Hypersensitivity Mediated by TRPV1 in the Rat Dorsal Root Ganglion

doi: 10.3389/fnmol.2021.663649

Figure Lengend Snippet: Molecular docking of A438079 with the TRPV1. (A) , top view. (B) , the docking pocket between A438079 and TRPV1. (C) , front view. (D) , A438079 does not form a hydrogen bond with TRPV1, and A438079 is in the middle.

Article Snippet: In this experiment, the rats were divided into a control group (control group), DNP model group (DNP group), P2X 7 receptor antagonist A438079 (S7705; Selleckchem)-treated DNP group (DNP + A438079 group), and normal saline (NS)-treated DNP group (DNP + NS group).

Techniques:

Journal: Frontiers in Immunology

Article Title: Extracellular ATP Limits Homeostatic T Cell Migration Within Lymph Nodes

doi: 10.3389/fimmu.2021.786595

Figure Lengend Snippet: ATP inhibits CCL19-induced chemotaxis but not S1P-induced chemotaxis of CD4 + T cells and CD8 + T cells via P2X7R. (A) CCL19-dependent T cell chemotaxis in the presence or absence of ATP was analyzed by Transwell assay. The result shown is representative of two independent experiments. Data represent the mean ± SD of triplicate wells. * p <0.05 by Student’s t -test. (B) The effect of ATP on CCL19-dependent T cell chemotaxis in the presence of ATP or apyrase-treated ATP in the lower well. The result shown is representative of three independent experiments. Data represent the mean ± SD of triplicate wells. * p <0.05 by Student’s t -test. (C) T cell chemotaxis in the presence of ATP and P2 receptor antagonists. CD4 + and CD8 + T cells were treated with A-438079 (P2X7R inhibitor; 10 μM), NF023 (P2X1R inhibitor; 10 μM), or 5-BDBD (P2X4R inhibitor; 10 μM) and subjected to the CCL19-dependent chemotaxis assay in the presence or absence of 100 μM ATP in the lower well. The result shown is representative of two independent experiments. Data represent the mean ± SD of triplicate wells. * p <0.05 by Student’s t -test. (D) Expression of P2X7R on CD3 + CD49d low T cells in LNs analyzed by flow cytometry. P2X7R is shown in black. Isotype control is shown in grey. The result shown is representative of at least three independent experiments. (E) An apparent contents map of ATP (ATP app ) generated as previously described and a confocal microscopic image of a serial LN section stained for CD4 (red), PNAd (green), and nucleus (blue). Images are representative of three individual experiments. (F) The effect of the P2X7R antagonist A-438079 on the intranodal retention of circulating lymphocytes. Data represent the mean ± SEM of five mice. * p <0.05 by Student’s t -test. (G) The effect of ATP on S1P-dependent T cell chemotaxis. S1P or CCL21 was added to the lower wells in the presence or absence of ATP at the indicated concentrations, and T cells were added to the upper wells. Relative cell migration was determined as described above. The result shown is representative of two independent experiments. Data represent the mean ± SD of triplicate wells. * p <0.05 by Student’s t -test; NS, not significant.

Article Snippet: A-438079 was purchased from Wako, and NF023 and 5-BDBD were from Tocris Bioscience.

Techniques: Chemotaxis Assay, Transwell Assay, Expressing, Flow Cytometry, Generated, Staining, Migration

Journal: Biology

Article Title: Activation of P2X7 Receptor Mediates the Abnormal Ovulation Induced by Chronic Restraint Stress and Chronic Cold Stress

doi: 10.3390/biology13080620

Figure Lengend Snippet: The expression of P2X7R in ovaries of chronic stress mice and the influence of P2X7R signaling on the ovulation of mice. After establishing the chronic stress model, immunohistochemistry and Western blotting were used to examine the expression change of P2X7R in ovarian tissues. ( A – C ) show the effect of chronic restraint stress (CRS) and chronic cold stress (CCS) on the expression of P2X7R in ovarian tissues. Superovulation was used to assess the ovulation rate of mice. ( D , E ) show the effect of A-438079 HCL, an antagonist of P2X7R signaling, on the superovulation rate of CRS and CCS mice. ( F ) shows the effect of BzATP and A-438079 HCL on the superovulation rate of normal healthy mice. * indicates p < 0.05 and ** indicates p < 0.01. Scale bar = 80 μm. The uncropped western blot figures were presented in .

Article Snippet: The mice were randomly divided into four groups (n = 6): control group, chronic restraint or cold stress group, A-438079 HCL (GC17212, GLPBIO, Montclair, CA, USA) group (P2X7R antagonist) and chronic restraint or cold stress+ A-438079 HCL group.

Techniques: Expressing, Immunohistochemistry, Western Blot

Journal: Biology

Article Title: Activation of P2X7 Receptor Mediates the Abnormal Ovulation Induced by Chronic Restraint Stress and Chronic Cold Stress

doi: 10.3390/biology13080620

Figure Lengend Snippet: The influence of P2X7R signaling on cumulus expansion in vitro and the expression of ovulation-related factors. ( A ) shows the effect of BzATP, an agonist of P2X7R signaling, on cumulus expansion induced by EGF. EGF served as positive control of cumulus expansion. Real-time PCR was used to detect the change in expression of ovulation-related factors. ( B ) shows the effect of A-438079 HCL, an antagonist of P2X7R signaling, on the expression of ovulation-related factors in granulosa cells. We found that NPPC was down-regulated. ( C ) shows the effect of BzATP and A-438079 HCL on NPPC mRNA expression. ( D ) shows the effect of BzATP and A-438079 HCL on CNP protein level. * indicates p < 0.05 and ** indicates p < 0.01. Scale bar = 100 μm.

Article Snippet: The mice were randomly divided into four groups (n = 6): control group, chronic restraint or cold stress group, A-438079 HCL (GC17212, GLPBIO, Montclair, CA, USA) group (P2X7R antagonist) and chronic restraint or cold stress+ A-438079 HCL group.

Techniques: In Vitro, Expressing, Positive Control, Real-time Polymerase Chain Reaction

Journal: Biology

Article Title: Activation of P2X7 Receptor Mediates the Abnormal Ovulation Induced by Chronic Restraint Stress and Chronic Cold Stress

doi: 10.3390/biology13080620

Figure Lengend Snippet: The influence of chronic stress on ovarian fibrosis. The changes in tissue fibrosis were detected by sirius red staining. ( A ) shows the representative sirius red staining of ovary in the control, chronic restraint stress (CRS) and chronic cold stress (CCS). ( B ) shows the expression of fibrosis-related factors, including TGF-β1 and MMP2 in chronic restraint stress (CRS) mice. ( C ) shows the expression of TGF-β1 and MMP2 in chronic cold stress (CCS) mice. ( D ) shows the effect of BzATP and A-438079 HCL on TGF-β1 and MMP2 expression in granulosa cells. * indicates p < 0.05 and ** indicates p < 0.01. Scale bar = 300 μm. The uncropped western blot figures were presented in .

Article Snippet: The mice were randomly divided into four groups (n = 6): control group, chronic restraint or cold stress group, A-438079 HCL (GC17212, GLPBIO, Montclair, CA, USA) group (P2X7R antagonist) and chronic restraint or cold stress+ A-438079 HCL group.

Techniques: Staining, Control, Expressing, Western Blot

Journal: Frontiers in Pharmacology

Article Title: Andrographolide Inhibits Mechanical and Thermal Hyperalgesia in a Rat Model of HIV-Induced Neuropathic Pain

doi: 10.3389/fphar.2018.00593

Figure Lengend Snippet: Effects of the Andro treatment on the expression of P2X7 mRNA and protein in L4–6 DRGs from each group. (A) Expression of P2X7 mRNA in the DRG was measured by RT-qPCR ( n = 8 per group). Expression in the gp120+ddC group was higher than that in the sham group ( p < 0.01). In gp120+ddC+andro rats, the expression was significantly lower than that in the gp120+ddC rats ( p < 0.01), which was same as the gp120+ddC+A438079-treated rats. No significant difference was found between the sham group and the control group or the control+andro group. Data are presented as the means ± SE. ∗∗ p < 0.01 compared to the sham group; ## p < 0.01 compared to the gp120+ddC-treated group. (B) Expression of P2X7 protein in the DRG was assessed by western blotting ( n = 8 for each group). P2X7 protein expression in the gp120+ddC-treated group was increased compared to the sham group ( p < 0.01). In gp120+ddC+andro rats, the expression was significantly lower than that in the gp120+ddC rats ( p < 0.01), which was same as the gp120+ddC+A438079-treated rats. No significant difference was found between the sham group and the control group or the control+andro group. Bar graphs show the ratio of the P2X7 receptor protein level to β-actin level in each group. Data are displayed as the means ± SE. ∗∗ p < 0.01 compared to the sham group; ## p < 0.01 compared to the gp120+ddC group.

Article Snippet: The rats were randomly arranged into different groups as follows (with eight rats in each group): the control group; control+andro group; the HIV-gp120 plus 2′,3′-dideoxycytidine (ddC) (intraperitoneal injection, i.p.) group (gp120+ddC group); the HIV-gp120 plus ddC rats treated with the Andro group (gp120+ddC+andro group); gp120+ddC+A438079 (P2X7 antagonist, AdooQ BioScience) group; and the sham operation group (sham group).

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Journal: Frontiers in Pharmacology

Article Title: Andrographolide Inhibits Mechanical and Thermal Hyperalgesia in a Rat Model of HIV-Induced Neuropathic Pain

doi: 10.3389/fphar.2018.00593

Figure Lengend Snippet: Treatment with Andro relieves mechanical and thermal hyperalgesia in gp120+ddC-treated rats. (A) The PWT in the gp120+ddC group was lower than that in the sham group ( p < 0.01). No significant difference was found between the sham group, control+andro group and the control group ( p > 0.05). In gp120+ddC+andro rats, the PWT was gradually higher than that in the gp120+ddC group ( p < 0.01), which was same as the gp120+ddC+A438079-treated rats. Each group consisted of eight rats ( n = 8 for each group). Data are displayed as the means ± SE. ∗ p < 0.05 compared to the sham group; ∗∗ p < 0.01 compared to the sham group; # p < 0.05 compared to the gp120+ddC-treated group; ## p < 0.01 compared to the gp120+ddC-treated group (two-way ANOVA test, F (7,336) = 35.46 for comparisons from different post-operation days, F (5,336) = 216.4 for comparisons from different groups of rats). (B) The PWL in the gp120+ddC group was lower than that in the sham group ( p < 0.01). No significant difference was found between the sham group, control+andro group and the control group ( p > 0.05). In gp120+ddC+andro rats, the PWL became gradually higher than that in the gp120+ddC group ( p < 0.05), which was same as the gp120+ddC+A438079-treated rats. Each group consisted of eight rats ( n = 8 for each group). Data are displayed as the means ± SE. ∗ p < 0.05 compared to the sham group; ∗∗ p < 0.01 compared to the sham group; # p < 0.05 compared to the gp120+ddC-treated group (two-way ANOVA test, F (7,336) = 56.81 for comparisons from different post-operation days, F (5,336) = 274 for comparisons from different groups of rats). (C) The PWT in gp120+ddC group rats was lower than that in the sham group rats ( p < 0.01), but there was no significant deference for the PWT between the gp120+ddC+andro group and gp120+ddC+andro+A438079 group ( p > 0.05). The PWT in the gp120+ddC+andro group rats was higher than that in the gp120+ddC group rats ( p < 0.01). Result in gp120+ddC+andro+A438079 group was the same as that in gp120+ddC+A438079 group ( p > 0.05). Each group consisted of eight rats ( n = 8 for each group). Data are displayed as the means ± SE. ∗ p < 0.05 compared to the sham group; ∗∗ p < 0.01 compared to the sham group; # p < 0.05 compared to the gp120+ddC-treated group (two-way ANOVA test, F (7,200) = 87.79 for comparisons from different post-operation days, F (4,200) = 131.8 for comparisons from different groups of rats). (D) The PWL in gp120+ddC group rats was lower than that in the sham group rats ( p < 0.01), but there was no significant deference for the PWL between the gp120+ddC+andro group and the gp120+ddC+andro+A438079 group ( p > 0.05). The PWL in the gp120+ddC+andro group rats was higher than that in the gp120+ddC group rats ( p < 0.01). Result in gp120+ddC+andro+A438079 group was the same as that in gp120+ddC+A438079 group ( p > 0.05). Each group consisted of eight rats ( n = 8 for each group). Data are displayed as the means ± SE. ∗ p < 0.05 compared to the sham group; ∗∗ p < 0.01 compared to the sham group; # p < 0.05 compared to the gp120+ddC-treated group (two-way ANOVA test, F (7,200) = 157.7 for comparisons from different post-operation days, F (4,200) = 189.9 for comparisons from different groups of rats).

Article Snippet: The rats were randomly arranged into different groups as follows (with eight rats in each group): the control group; control+andro group; the HIV-gp120 plus 2′,3′-dideoxycytidine (ddC) (intraperitoneal injection, i.p.) group (gp120+ddC group); the HIV-gp120 plus ddC rats treated with the Andro group (gp120+ddC+andro group); gp120+ddC+A438079 (P2X7 antagonist, AdooQ BioScience) group; and the sham operation group (sham group).

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Andrographolide Inhibits Mechanical and Thermal Hyperalgesia in a Rat Model of HIV-Induced Neuropathic Pain

doi: 10.3389/fphar.2018.00593

Figure Lengend Snippet: Treatment with Andro reduces the upregulated co-expression values of the P2X7 receptor and GFAP in the DRG of gp120+ddC-treated rats. (A) The co-expression of P2X7 receptor and GFAP was analyzed on the 18–20th day after surgery. Co-expression of the P2X7 receptor and GFAP in the gp120+ddC group was higher than that in the sham group. No difference was found between control+andro and sham rats. Treatment with Andro reduced the co-expression values of the P2X7 receptor and GFAP in gp120+ddC+andro-treated rats compared with gp120+ddC-treated rats that did not receive Andro treatment ( n = 8 for each group). Scale bar: 30 μm. (B) Histogram showed that the IOD value for co-expression of the P2X7 receptor and GFAP in the DRG. (C) Histogram showed that the number of neurons surrounded with GFAP- and P2X7-positive SGCs in the DRG SCGs. The data are shown as the mean ± SD. ∗∗ p < 0.01, compared to the sham group; ## p < 0.01, compared to the gp120+ddC group ( n = 8 for each group).

Article Snippet: The rats were randomly arranged into different groups as follows (with eight rats in each group): the control group; control+andro group; the HIV-gp120 plus 2′,3′-dideoxycytidine (ddC) (intraperitoneal injection, i.p.) group (gp120+ddC group); the HIV-gp120 plus ddC rats treated with the Andro group (gp120+ddC+andro group); gp120+ddC+A438079 (P2X7 antagonist, AdooQ BioScience) group; and the sham operation group (sham group).

Techniques: Expressing

Journal: Frontiers in Pharmacology

Article Title: Andrographolide Inhibits Mechanical and Thermal Hyperalgesia in a Rat Model of HIV-Induced Neuropathic Pain

doi: 10.3389/fphar.2018.00593

Figure Lengend Snippet: Effects of the Andro treatment on the expression of TNF-α-R, IL-1β, and IL-10 protein in L4–6 DRGs. (A) The expression of IL-1β protein in the DRG of the gp120+ddC group was increased compared to the sham group ( p < 0.01). The expression of IL-1β protein in the gp120+ddC+andro group was decreased compared to the gp120+ddC group ( p < 0.01). The bar graphs show the ratio of the IL-1β protein level to β-actin level in each group, respectively. The data are shown as the mean ± SD. p < 0.01, compared to the sham group; ## p < 0.01, compared to the gp120+ddC group ( n = 8 for each group). (B) The expression of TNF-α-R protein in the DRG of the gp120+ddC group was increased compared to the sham group ( p < 0.01). The expression of TNF-α-R protein in the gp120+ddC+andro group was decreased compared to the gp120+ddC group ( p < 0.01). The bar graphs show the ratio of the TNF-α-R protein level to β-actin level in each group, respectively. The data are shown as the mean ± SD. p < 0.01, compared to the sham group; ## p < 0.01, compared to the gp120+ddC group ( n = 8 for each group). (C) The expression of IL-10 protein in the DRG of the gp120+ddC group was decreased compared to the sham group ( p < 0.01). The expression of IL-10 protein in the DRG of the gp120+ddC+andro group was increased compared to the gp120+ddC group ( p < 0.01). The bar graphs show the ratio of the IL-10 protein level to β-actin level in each group, respectively. The data are shown as the mean ± SD. ∗∗ p < 0.01, compared to the sham group; ## p < 0.01, compared to the gp120+ddC group ( n = 8 for each group).

Article Snippet: The rats were randomly arranged into different groups as follows (with eight rats in each group): the control group; control+andro group; the HIV-gp120 plus 2′,3′-dideoxycytidine (ddC) (intraperitoneal injection, i.p.) group (gp120+ddC group); the HIV-gp120 plus ddC rats treated with the Andro group (gp120+ddC+andro group); gp120+ddC+A438079 (P2X7 antagonist, AdooQ BioScience) group; and the sham operation group (sham group).

Techniques: Expressing

Journal: Frontiers in Pharmacology

Article Title: Andrographolide Inhibits Mechanical and Thermal Hyperalgesia in a Rat Model of HIV-Induced Neuropathic Pain

doi: 10.3389/fphar.2018.00593

Figure Lengend Snippet: Effects of Andro treatment on the expression of ERK1/2 and p-ERK1/2 in L4–6 DRGs. (A) The expression levels of ERK1/2 and p-ERK1/2 in the DRG were analyzed using western blotting. (B) The IOD ratio of ERK1/2 to β-actin was not significantly different among the four groups: the gp120+ddC group, the sham group, the control+andro group, and the gp120+ddC+andro group (one-way ANOVA test, F (3,28) = 0.05357, p = 0.9833 for comparisons from different groups). (C) The bar graphs show that the IOD ratio of p-ERK1/2 to ERK1/2 in the gp120+ddC group was higher than that in the sham group ( n = 8) and the IOD ratio of p-ERK1/2 to ERK1/2 in the gp120+ddC+andro-treated rats was significantly lower than that in the gp120+ddC group ( p < 0.01, n = 8 for each group). The data represent the mean ± SD, n = 8. ∗∗ p < 0.01 compared to the sham group; ## p < 0.01 compared to the gp120+ddC group.

Article Snippet: The rats were randomly arranged into different groups as follows (with eight rats in each group): the control group; control+andro group; the HIV-gp120 plus 2′,3′-dideoxycytidine (ddC) (intraperitoneal injection, i.p.) group (gp120+ddC group); the HIV-gp120 plus ddC rats treated with the Andro group (gp120+ddC+andro group); gp120+ddC+A438079 (P2X7 antagonist, AdooQ BioScience) group; and the sham operation group (sham group).

Techniques: Expressing, Western Blot

Journal: Frontiers in Pharmacology

Article Title: Andrographolide Inhibits Mechanical and Thermal Hyperalgesia in a Rat Model of HIV-Induced Neuropathic Pain

doi: 10.3389/fphar.2018.00593

Figure Lengend Snippet: MOE score of P2X7 protein and andrographolide (kcal/mol).

Article Snippet: The rats were randomly arranged into different groups as follows (with eight rats in each group): the control group; control+andro group; the HIV-gp120 plus 2′,3′-dideoxycytidine (ddC) (intraperitoneal injection, i.p.) group (gp120+ddC group); the HIV-gp120 plus ddC rats treated with the Andro group (gp120+ddC+andro group); gp120+ddC+A438079 (P2X7 antagonist, AdooQ BioScience) group; and the sham operation group (sham group).

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Andrographolide Inhibits Mechanical and Thermal Hyperalgesia in a Rat Model of HIV-Induced Neuropathic Pain

doi: 10.3389/fphar.2018.00593

Figure Lengend Snippet: Molecular docking of rP2X7 protein and Andro. Simulation modeling of Andro docking with P2X7 protein was simulated by computer. (A) Front and aerial view showed that the best docking position between Andro and rP2X7. The docking position was in the outside of the cell membrane and at the waist of the protein structure, which is near the ATP docking pocket. (B) Showed that the best docking pocket between Andro and rP2X7. The ligand, Andro represented in yellow, and P2X7 protein’s active binding pocket displayed in a variety of colors. (C) Indicates a possible hydrogen bond formed between the connection residues (GLN143) and ligand. The data showed that Andro could interact with the P2X7 protein.

Article Snippet: The rats were randomly arranged into different groups as follows (with eight rats in each group): the control group; control+andro group; the HIV-gp120 plus 2′,3′-dideoxycytidine (ddC) (intraperitoneal injection, i.p.) group (gp120+ddC group); the HIV-gp120 plus ddC rats treated with the Andro group (gp120+ddC+andro group); gp120+ddC+A438079 (P2X7 antagonist, AdooQ BioScience) group; and the sham operation group (sham group).

Techniques: Binding Assay