Journal: Cells

Article Title: Liraglutide Potently Protects Against Streptozotocin-Induced Islet Injury Associated with Inhibition of HMGB1 Release

doi: 10.3390/cells15131203

Figure Lengend Snippet: Liraglutide reduces STZ-induced TLR4 expression in mouse pancreas and inflammatory factor levels in vivo. ( A ) Immunohistochemistry detection of the expression of HMGB1 receptor AGER, TLR4, and TLR2 in pancreatic tissues. ( B ) ELISA detection of IL-1β expression, by group ( n = 3). ( C ) ELISA detection of IFN-γ expression, by group ( n = 3). ( D ) ELISA detection of CXCL10 expression, by group ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: ELISA kits were used to detect HMGB1 ( F10620 , Xitang, Changsha, China), insulin (CSB-E05071m, CUSABIO, Wuhan, China), IL-1β (RK00006, ABclonal, Wuhan, China), IFN-γ (RK00019, ABclonal, China), and CXCL10 (RK00056, ABclonal, Wuhan, China).

Techniques: Expressing, In Vivo, Immunohistochemistry, Enzyme-linked Immunosorbent Assay

Journal: Acta pharmaceutica Sinica. B

Article Title: Macrophage P2Y 6 R activation aggravates psoriatic inflammation through IL-27-mediated Th1 responses.

doi: 10.1016/j.apsb.2024.06.008

Figure Lengend Snippet: Figure 2 Macrophage P2Y6R mediated Th1 type psoriasis-like inflammation via p-JAK2/p-STAT1/T-bet signaling. (A) Flow diagram. (B, C) FACs and the statistical analysis of IFNgþ CD4þ T-cells (n Z 3). (D) IFNg concentration after WT or P2Y6R/ mice BMDM supernatant inducing Naı¨ve CD4þ T-cells (n Z 4). (E) The mRNA expression of the Th1-associated cytokines, Ifng, Cxcl9, Cxcl10, and Cxcl11 were detected by RT-qPCR (n Z 3). All mRNA levels were normalized to GAPDH. (F) Linear regression analysis indicated a positive correlation between P2ry6 and Stat1 in GSE181318 and GSE14905. (G) GSEA analysis of GSE50400 showed JNK/STAT signaling pathways were up-regulated in IMQ-induced mice. (H) Top 15 putative transcription factors based on ChEA3. (I, J) The protein expression levels of p-JAK2, p-STAT1, and T-bet in T cells. Normal protein levels were normalized to b-actin, the values for the phosphorylated forms of proteins were normalized to phosphorylation-independent levels of the same protein (n Z 4). (K) CD4þ Naı¨ve T cells supernatant IFNg concentration after WT or P2Y6R/

Article Snippet: IL-27 and IFNg were detected by mouse IL-27 ELISA kit (EMC115.96, Neobioscience Technology, Shenzhen, China) and mouse IFNg ELISA kit (EK280HS, MULTI Sciences, Hangzhou, China) according to the manufacturer’s instructions.

Techniques: Concentration Assay, Expressing, Quantitative RT-PCR, Protein-Protein interactions, Phospho-proteomics

Journal: Acta pharmaceutica Sinica. B

Article Title: Macrophage P2Y 6 R activation aggravates psoriatic inflammation through IL-27-mediated Th1 responses.

doi: 10.1016/j.apsb.2024.06.008

Figure Lengend Snippet: Figure 3 P2Y6R enhances IL-27 released from macrophages. (A) Top15 BPs were shown through GO analysis of DEGs. (B) Linear regression analysis indicated a positive correlation between P2ry6 and Il-27 in GSE181318 and GSE14905. (C) Flow diagram. (D) The IL-27 concentration in the supernatants of P2Y6R/ or WT BMDMs induced by IMQ and UDP (n Z 4). (E) Flow diagram. (FeG) The proportion of Th1 cells differentiated from IL27ra/ Naı¨ve T cells has no significant variability between WT BMDMs and P2Y6R/ BMDMs supernatant stimulation tested by FACs (n Z 3). (H) WT or IL27ra/ mice CD4þ Naı¨ve T cells supernatant IFNg concentration after WT or P2Y6R/ mice BMDM supernatant inducing (n Z 4). (I) Changes in BMDMs intracellular Ca2þ were measured using the Ca2þ detection Fluo-4AM molecular probes. (J) Quantitative analysis of fluorescence intensity. (KeL) Protein expression in different groups. Normal protein levels were normalized to b-actin, the values for the phosphorylated forms of proteins were normaliszd to phosphorylation-independent levels of the same protein (n Z 4). (M) The IL-27 concentration in the supernatant. Data are expressed as mean SEM. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: IL-27 and IFNg were detected by mouse IL-27 ELISA kit (EMC115.96, Neobioscience Technology, Shenzhen, China) and mouse IFNg ELISA kit (EK280HS, MULTI Sciences, Hangzhou, China) according to the manufacturer’s instructions.

Techniques: Concentration Assay, Expressing, Phospho-proteomics

Journal: Acta pharmaceutica Sinica. B

Article Title: Macrophage P2Y 6 R activation aggravates psoriatic inflammation through IL-27-mediated Th1 responses.

doi: 10.1016/j.apsb.2024.06.008

Figure Lengend Snippet: Figure 5 Increase of IL-27 restores psoriatic inflammation in Lyz2cre P2Y6Rfl/flmice. (A) Flow diagram. (B) Representative psoriatic plaque pictures of mice. (C) Pictures of the spleen. (D) Spleen index of all groups. (E, F) Skin thickness, erythema and scaling PASI score, H&E staining (n Z 3), and Ki67 staining (n Z 3) of IMQ-induced Lyz2creP2Y6Rfl/fland P2Y6Rfl/flmice skin treated with Saline or IL-27. (G) The mRNA expression levels of Th1-related cytokines in skin tissues from IMQ-induced mice treated with Saline or IL-27 (n Z 3). All mRNA levels were normalized to GAPDH mRNA. (H, I) Flow analysis on Th1 cells from skin and spleens of all groups. IL-27 reversed the decrease of Th1 cells (CD4þ IFNgþ) proportion in IMQ-treated Lyz2creP2Y6Rfl/flmice. (J, K) Quantitative analysis of protein expression in mice skin. Normal protein levels were normalized to b-actin, and the values for the phosphorylated forms of proteins were normalized to phosphorylation-independent levels of the same protein (n Z 4). (L) Serum and skin concentrations of IL-27 and IFNg detected by ELISA assay (n Z 4). Data are expressed as mean SEM. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: IL-27 and IFNg were detected by mouse IL-27 ELISA kit (EMC115.96, Neobioscience Technology, Shenzhen, China) and mouse IFNg ELISA kit (EK280HS, MULTI Sciences, Hangzhou, China) according to the manufacturer’s instructions.

Techniques: Staining, Saline, Expressing, Phospho-proteomics, Enzyme-linked Immunosorbent Assay

Journal: Acta pharmaceutica Sinica. B

Article Title: Macrophage P2Y 6 R activation aggravates psoriatic inflammation through IL-27-mediated Th1 responses.

doi: 10.1016/j.apsb.2024.06.008

Figure Lengend Snippet: Figure 6 A novel P2Y6R inhibitor FS-6 with excellent activity and binding affinity. (A) The overall structure of P2Y6R with FS-6 bound. (B) Chemical modification from A6 to FS-6 and the possible binding sites between FS-6 and P2Y6R. (C) Flow diagram of IC50 value and binding activity of FS-6. (D) IC50 curve of FS-6 was determined by ELISA (n Z 6). (E) Direct binding between FS-6 and P2Y6R. (FeI) The binding between FS-6 and P2Y6R was evaluated by CETSA in P2Y6R-HEK293 cells (n Z 4). (J‒M) The dose-dependent binding of FS-6 to P2Y6R was evaluated by DARTs (n Z 4). (N) Arg266Gly was verified as the binding site of FS-6 and P2Y6R to detect IP3 concentration (n Z 4). Data are expressed as mean SEM. *P < 0.05, **P < 0.01, ***P < 0.001. ns, not significant.

Article Snippet: IL-27 and IFNg were detected by mouse IL-27 ELISA kit (EMC115.96, Neobioscience Technology, Shenzhen, China) and mouse IFNg ELISA kit (EK280HS, MULTI Sciences, Hangzhou, China) according to the manufacturer’s instructions.

Techniques: Activity Assay, Binding Assay, Enzyme-linked Immunosorbent Assay, Concentration Assay

Journal: Acta pharmaceutica Sinica. B

Article Title: Macrophage P2Y 6 R activation aggravates psoriatic inflammation through IL-27-mediated Th1 responses.

doi: 10.1016/j.apsb.2024.06.008

Figure Lengend Snippet: Figure 8 FS-6 ameliorates IMQ-induced psoriasis in mice. (A) Experimental scheme for assessment of psoriasiform dermatitis (n Z 12). (B) Representative skin pictures. (C) Picture of spleen and (D) spleen index from all groups (n Z 12). (E) PASI score (n Z 12). (F) Serum and skin concentrations of IL-27 and IFNg (n Z 4). (G) FS-6 reduced epidermal thickening, keratinocyte proliferation, and co-localization of macrophages and P2Y6R (n Z 3). (H, I) Flow analysis on Th1 cells from skin and spleens of mice (n Z 3). Data are expressed as mean SEM. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: IL-27 and IFNg were detected by mouse IL-27 ELISA kit (EMC115.96, Neobioscience Technology, Shenzhen, China) and mouse IFNg ELISA kit (EK280HS, MULTI Sciences, Hangzhou, China) according to the manufacturer’s instructions.

Techniques: