Journal: Frontiers in Immunology

Article Title: Targeting Purinergic Receptor P2RX1 Modulates Intestinal Microbiota and Alleviates Inflammation in Colitis

doi: 10.3389/fimmu.2021.696766

Figure Lengend Snippet: Purinergic receptor P2RX1 expression is increased in activated colitis. (A) Expression of purinergic receptors were analyzed in two Gene Expression Omnibus (GEO) datasets (GSE59071 and GSE53306) containing expression profile of actively inflamed mucosa from colitis patients, and one GEO dataset (GSE22307) containing expression profile of colon tissues from DSS-induced mouse colitis. Venn diagram of significantly upregulated and downregulated genes was shown. (B, C) Expression profiles of P2RX1 in the GSE59071, GSE53306, GSE22307, and GSE16879 datasets. *P < 0.05, **P < 0.01, and ***P < 0.001.

Article Snippet: For P2RX1 staining, anti-P2RX1 (Alomone, APR-022, 1:80) antibody followed by an anti-rabbit-PE secondary antibody (Abcam, 1:200) was used.

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: Targeting Purinergic Receptor P2RX1 Modulates Intestinal Microbiota and Alleviates Inflammation in Colitis

doi: 10.3389/fimmu.2021.696766

Figure Lengend Snippet: P2RX1 ablation relieves DSS-induced mouse colitis. (A, B) WT and P2rx1 −/− mice were treated with 2% DSS for 7 days. Weight loss (A) and disease activity index (DAI) (B) were calculated (n = 6 per group). (C) Colon length of WT and P2rx1 −/− mice was measured at days 0 and 7. Representative colon tissues at day 7 were shown (n = 6 per group). Scale bar is 1 cm. (D) Histologic score of WT and P2rx1 −/− mice at days 0 and 7 was determined according to pathological examinations (n = 6 per group). (E) WT and P2rx1 −/− mice were exposed to 3% DSS for 7 days, and survival status was monitored for 25 days (n = 10 per group). *P < 0.05, **P < 0.01, and ***P < 0.001.

Article Snippet: For P2RX1 staining, anti-P2RX1 (Alomone, APR-022, 1:80) antibody followed by an anti-rabbit-PE secondary antibody (Abcam, 1:200) was used.

Techniques: Activity Assay

Journal: Frontiers in Immunology

Article Title: Targeting Purinergic Receptor P2RX1 Modulates Intestinal Microbiota and Alleviates Inflammation in Colitis

doi: 10.3389/fimmu.2021.696766

Figure Lengend Snippet: P2RX1 ablation restricts inflammatory responses in DSS-induced mouse colitis. (A) WT and P2rx1 −/− mice were treated with 2% DSS for 7 days. At days 0, 7, and 12, colon tissues were harvested, and RNA sequencing was performed. Differential analysis was analyzed, with P <0.001 and fold change >4 being considered as significantly varied (n = 3 for D0 P2rx1 −/− group, and n = 1 for the rest of the groups). (B) KEGG pathway analysis was performed to compare the functional enrichment genes of WT and P2rx1 −/− mice at day 7. (C) A heatmap of inflammation-associated genes was shown. (D) Inflammation-associated genes were detected by RT-qPCR (n = 4 per group). (E) Neutrophils and macrophages were analyzed by flow cytometry (n = 4 per group). **P < 0.01 and ***P < 0.001.

Article Snippet: For P2RX1 staining, anti-P2RX1 (Alomone, APR-022, 1:80) antibody followed by an anti-rabbit-PE secondary antibody (Abcam, 1:200) was used.

Techniques: RNA Sequencing Assay, Functional Assay, Quantitative RT-PCR, Flow Cytometry

Journal: Frontiers in Immunology

Article Title: Targeting Purinergic Receptor P2RX1 Modulates Intestinal Microbiota and Alleviates Inflammation in Colitis

doi: 10.3389/fimmu.2021.696766

Figure Lengend Snippet: Intestinal microbiota is altered in DSS-treated P2rx1 −/− mice. (A) WT and P2rx1 −/− mice were treated with 2% DSS for 7 days. At days 0, 7 and 12, fecal microbiota was quantified using 16S rDNA sequencing. Principal component analysis (PCA) was performed based on the operational taxonomic units (OTUs) composition (n = 6 for D0 WT and P2rx1 −/− groups, n = 3 for the rest groups). (B) Relative abundance of bacterial phylum in the fecal microbiota was analyzed. (C) Detailed comparison of bacterial genus in WT and P2rx1 −/− mice was performed. (D) Functional metagenomics prediction analysis based on the result of 16S rDNA gene sequencing using PICRUSt1 was performed. (E) Fecal indole-3-acetic acid (IAA) and indole at day 7 were detected. IL-22 mRNA and protein at day 0 and day 7 were determined (n = 4 per group). **P < 0.01.

Article Snippet: For P2RX1 staining, anti-P2RX1 (Alomone, APR-022, 1:80) antibody followed by an anti-rabbit-PE secondary antibody (Abcam, 1:200) was used.

Techniques: Sequencing, Functional Assay

Journal: Frontiers in Immunology

Article Title: Targeting Purinergic Receptor P2RX1 Modulates Intestinal Microbiota and Alleviates Inflammation in Colitis

doi: 10.3389/fimmu.2021.696766

Figure Lengend Snippet: Inhibition of P2RX1 promotes the efficiency of anti-TNF-α therapy in mouse colitis. (A, B) WT mice were treated with 2% DSS for 7 days. Anti-TNF-α monoclonal antibody (mAb) (0.1 mg/day) or/and P2RX1 inhibitor (NF449, 10 mg/kg/day) was/were administrated for 7 days. Weight loss (A) and disease activity index (DAI) (B) were calculated (n = 6 per group). Statistical analyses were performed at day 7. (C) At day 7, RT-qPCR was performed to detect inflammatory cytokine expression (n = 4 per group). (D) At day 7, colonic neutrophils were determined by cytometry (n = 4 per group). *P < 0.05, **P < 0.01, and ***P < 0.001.

Article Snippet: For P2RX1 staining, anti-P2RX1 (Alomone, APR-022, 1:80) antibody followed by an anti-rabbit-PE secondary antibody (Abcam, 1:200) was used.

Techniques: Inhibition, Activity Assay, Quantitative RT-PCR, Expressing, Cytometry