Journal: ImmunoTargets and Therapy

Article Title: Recombinant Antithrombin Alleviated Pulmonary Injury and Inflammation in LPS-Induced ARDS by Inhibiting IL17a/NF-κB Signaling

doi: 10.2147/ITT.S502925

Figure Lengend Snippet: rAT reduced pulmonary inflammation in LPS-induced ARDS model mice. ( A–D ) The levels of inflammatory factors, including IL-6 ( A ), TNF-α ( B ), IL-8 ( C ), and hs-CRP ( D ), were decreased in the serum of the rAT-treated group, as detected by ELISA. ( E ) Immunohistochemical staining for F4/80 revealed that rAT treatment reduced the proportion of macrophages in the lung tissue of LPS-induced ARDS model mice. ( F ) Immunohistochemical staining for MRCI, a marker of M2 macrophages, showed that rAT treatment increased the proportion of M2 macrophages in the lung tissue of LPS-induced ARDS model mice. ( G ) Immunohistochemical staining for Ly6G, a marker of neutrophils, showed that rAT treatment reduced the proportion of neutrophils in the lung tissue of LPS-induced ARDS model mice. All the data are presented as the means ± SDs of three independent experiments. One-way ANOVA, *p < 0.05, **p < 0.01, ***p < 0.001, ns, not significant; scale bar, 50 μm.

Article Snippet: A mouse CXCL15 ELISA Kit (E-EL-M0269) and a mouse hs-CRP ELISA Kit (E-EL-M0677) were purchased from Elabscience (Wuhan, China).

Techniques: Enzyme-linked Immunosorbent Assay, Immunohistochemical staining, Staining, Marker

Journal: ImmunoTargets and Therapy

Article Title: Recombinant Antithrombin Alleviated Pulmonary Injury and Inflammation in LPS-Induced ARDS by Inhibiting IL17a/NF-κB Signaling

doi: 10.2147/ITT.S502925

Figure Lengend Snippet: The efficacy of rAT in mitigating lung injury, suppressing the immune response, and inhibiting the activation of the NF-κB signaling pathway in LPS-induced ARDS mice were diminished by the administration of IL-17a. ( A ) ELISA results demonstrated that the administration of IL17a inhibited the ability of rAT to reduce inflammatory factors, including IL-6, TNF-α, and IL-8, in the serum of LPS-induced ARDS mice. ( B ) The analysis of the wet/dry weight ratio of the lung tissue revealed that the administration of IL17a counteracted the ability of rAT to alleviate pulmonary exudation in LPS-induced ARDS mice. ( C ) The administration of IL17a did not significantly affect the ability of rAT to reduce the number of cells in the BALF of LPS-induced ARDS mice. ( D ) The administration of IL17a attenuated the ability of rAT to reduce the concentrations of proteins in the BALF of LPS-induced ARDS mice. ( E ) Real-time PCR results showed that the administration of IL17a blocked the ability of rAT to downregulate the expression of target genes in the IL17a/NF-κB signaling pathway. ( F ) The protein levels of the NF-κB signaling pathway were assessed by Western blotting, and gray intensity analysis of the blots showed that the administration of IL17a in LPS-induced ARDS mice counteracted the ability of rAT to suppress the phosphorylation of IκBα, IKKα/β, and P65. The data are expressed as the means ± SDs (n=3 in each group). One-way ANOVA, *p < 0.05, **p < 0.01, ***p < 0.001, and ns not significant.

Article Snippet: A mouse CXCL15 ELISA Kit (E-EL-M0269) and a mouse hs-CRP ELISA Kit (E-EL-M0677) were purchased from Elabscience (Wuhan, China).

Techniques: Activation Assay, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Phospho-proteomics

Journal: International journal of molecular sciences

Article Title: Modulation of the Cardiovascular Risk in Type 1 Diabetic Rats by Endurance Training in Combination with the Prebiotic Xylooligosaccharide.

doi: 10.3390/ijms251810027

Figure Lengend Snippet: Figure 4. Study design. BP—blood pressure, HR—heart rate, MTE—maximum time to ex- haustion, AC—abdominal circumference, T-AOC—total antioxidant capacity, Il-6—interleukin-6, hsCRP—high-sensitive C-reactive protein, NADH + H+—nicotinamide adenine dinucleotide hydride, CoQ—coenzyme Q, LDH—lactate dehydrogenase, SDH—succinate dehydrogenase.

Article Snippet: Serum T-AOC, IL-6 and hsCRP were analyzed on an enzyme-linked immunosorbent assay (ELISA) microplate reader HumanReader.HS, HUMAN (Wiesbaden, Germany) via commercially available kits [Rat Total antioxidant capacity, T-AOC Elisa kit, Nanjing Pars Biochem CO., Ltd., Nanjing, China; Rat IL-6 (Interleukin 6) ELISA Kit, Elabscience Biotechnology Inc., Houston, TX, USA; Rat hsCRP (high-sensitivity, C-Reactive Protein) Elabscience Biotechnology Inc., Houston, TX, USA].

Techniques:

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:

Journal: Inflammation

Article Title: CXCL12 Promotes Peripheral Nerve Injury Repair by Inhibiting the Ferroptosis-Inflammation Axis via the ERK/Nrf2 Pathway

doi: 10.1007/s10753-026-02453-2

Figure Lengend Snippet: CXCL12 inhibits NF-κB signaling pathway activation and inflammatory factor secretion in SCs. ( A ) Western blot analysis of NF-κB, p-NF-κB, IκBα, and p-IκBα protein expression levels in SCs after CXCL12 treatment. ( B - C ) Quantification of gray values for the NF-κB, p-NF-κB, κBα, and p-IκBα proteins. ( D - E ) Levels of IL-1β and TNF-α in SCs after CXCL12 treatment. The data are presented as the means ± SDs, n=3, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: An enzyme-linked immunosorbent assay was used to measure the concentrations of the inflammatory factors IL-1β (Elabscience, E-HSEL-R0002, China) and TNF-α (Elabscience, E-HSEL-R0001, China) in the cell culture supernatant (centrifuged at 3000×g for 15 min).

Techniques: Activation Assay, Western Blot, Expressing

Journal: Inflammation

Article Title: CXCL12 Promotes Peripheral Nerve Injury Repair by Inhibiting the Ferroptosis-Inflammation Axis via the ERK/Nrf2 Pathway

doi: 10.1007/s10753-026-02453-2

Figure Lengend Snippet: CXCL12 inhibits the inflammatory response in SCs via the NF-κB signaling pathway. ( A ) Western blot analysis of NF-κB, p-NF-κB, IκBα, and p-IκBα protein expression levels in SCs after NF-κB overexpression. ( B - C ) Quantification of the gray values for the NF-κB, p-NF-κB, IκBα, and p-IκBα proteins. ( D - E ) Levels of IL-1β and TNF-α in SCs after NF-κB overexpression. The data are presented as the means ± SDs, n=3, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns, not significant

Article Snippet: An enzyme-linked immunosorbent assay was used to measure the concentrations of the inflammatory factors IL-1β (Elabscience, E-HSEL-R0002, China) and TNF-α (Elabscience, E-HSEL-R0001, China) in the cell culture supernatant (centrifuged at 3000×g for 15 min).

Techniques: Western Blot, Expressing, Over Expression

Journal: Inflammation

Article Title: CXCL12 Promotes Peripheral Nerve Injury Repair by Inhibiting the Ferroptosis-Inflammation Axis via the ERK/Nrf2 Pathway

doi: 10.1007/s10753-026-02453-2

Figure Lengend Snippet: CXCL12 inhibits SC ferroptosis, thereby mitigating the inflammatory response. ( A ) Western blot analysis of the expression levels of ferroptosis-related proteins ACSL4, GPX4, and FSP1 in SCs after FAC treatment. ( B - D ) Densitometric analysis of proteins ACSL4, GPX4, and FSP1. ( E , F ) Fluorescence images of ROS and Fe 2+ in SCs after FAC treatment, Scale bar = 50µm. ( G , H ) Quantification of the relative fluorescence intensity of ROS and Fe 2+ . ( I , J ) Expression levels of MDA and GSH in SCs after FAC treatment. ( K ) Western blot analysis of the expression levels of NF-κB, p-NF-κB, IκBα, and p-IκBα in SCs after FAC treatment. ( L , M ) Densitometric analysis of proteins NF-κB, p-NF-κB, IκBα, and p-IκBα. ( N , O ) Expression levels of IL-1β and TNF-α in SCs after FAC treatment. The data are presented as the means ± SDs, n=3, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns, not significant

Article Snippet: An enzyme-linked immunosorbent assay was used to measure the concentrations of the inflammatory factors IL-1β (Elabscience, E-HSEL-R0002, China) and TNF-α (Elabscience, E-HSEL-R0001, China) in the cell culture supernatant (centrifuged at 3000×g for 15 min).

Techniques: Western Blot, Expressing, Fluorescence

Journal: Inflammation

Article Title: CXCL12 Promotes Peripheral Nerve Injury Repair by Inhibiting the Ferroptosis-Inflammation Axis via the ERK/Nrf2 Pathway

doi: 10.1007/s10753-026-02453-2

Figure Lengend Snippet: CXCL12 inhibits NF-κB signaling pathway activation and inflammatory factor secretion in the sciatic nerve. ( A ) Western blot analysis of NF-κB, p-NF-κB, IκBα, and p-IκBα protein expression levels in the sciatic nerve after CXCL12 treatment. ( B - C ) Quantification of the gray values for the NF-κB, p-NF-κB, IκBα, and p-IκBα proteins. ( D - E ) Levels of IL-1β and TNF-α in the sciatic nerve after CXCL12 treatment. The data are presented as the means ± SDs, n=3, **p < 0.01, ***p < 0.001, **** p < 0.0001

Article Snippet: An enzyme-linked immunosorbent assay was used to measure the concentrations of the inflammatory factors IL-1β (Elabscience, E-HSEL-R0002, China) and TNF-α (Elabscience, E-HSEL-R0001, China) in the cell culture supernatant (centrifuged at 3000×g for 15 min).

Techniques: Activation Assay, Western Blot, Expressing

Journal: Inflammation

Article Title: CXCL12 Promotes Peripheral Nerve Injury Repair by Inhibiting the Ferroptosis-Inflammation Axis via the ERK/Nrf2 Pathway

doi: 10.1007/s10753-026-02453-2

Figure Lengend Snippet: CXCL12 inhibits sciatic nerve ferroptosis, thereby mitigating the inflammatory response. ( A ) Western blot analysis of ferroptosis-related protein expression levels (ACSL4, GPX4, FSP1) in the sciatic nerve after FAC treatment. ( B - D ) Quantification of the gray values for the ACSL4, GPX4, and FSP1 proteins. ( E ‒ G ) Levels of Fe 2+ , MDA, and GSH in the sciatic nerve after FAC treatment. ( H ) Western blot analysis of NF-κB, p-NF-κB, IκBα and p-IκBα protein expression levels in the sciatic nerve after FAC treatment. ( I , J ) Quantification of the gray values for the NF-κB, p-NF-κB, IκBα and p-IκBα proteins. (K, L) Levels of IL-1β and TNF-α in the sciatic nerve after FAC treatment. The data are presented as the means ± SDs, n=3, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: An enzyme-linked immunosorbent assay was used to measure the concentrations of the inflammatory factors IL-1β (Elabscience, E-HSEL-R0002, China) and TNF-α (Elabscience, E-HSEL-R0001, China) in the cell culture supernatant (centrifuged at 3000×g for 15 min).

Techniques: Western Blot, Expressing