Journal: Inflammation

Article Title: Targeting Cold-Inducible RNA-Binding Protein Attenuates Pancreatic Fibrosis by Suppressing Pyroptosis in Chronic Pancreatitis

doi: 10.1007/s10753-026-02490-x

Figure Lengend Snippet: Inflammation and pyroptosis were increased in CP mice. Cerulein-CP was induced by 6 IP injections of cerulein (50 µg/kg/body weight) twice a week for 10 weeks. L-arginine-CP was induced by 2 hourly IP injections of 4.0 g/kg L-arginine twice a week for 10 weeks. The animals were sacrificed at 10 weeks after the first injection of L-arginine or cerulein. Blood and tissue samples were collected. ( A-C ) Representative photos and quantitative of F4/80 and CD11b staining; ( D ) Serum IL-6 level; ( E ) Serum TNF-α level; ( F & G ) Western blot analysis and quantitative of the expression of NLRP3 in the pancreas; ( H ) Serum IL-18 level; ( I ) Serum IL-1β level; ( J ) Correlation analysis of serum IL-18 and Masson positive aera; ( K ) Correlation analysis of serum IL-1β and Masson positive aera; ( L ) Correlation analysis of serum CIRP and F4/80 positive cells; ( M ) Correlation analysis of serum CIRP and CD11b positive cells; ( N ) Correlation analysis of serum CIRP and F4/80 positive cells; ( O ) Correlation analysis of serum CIRP and CD11b positive cells; ( R ) Correlation analysis of serum CIRP and IL-18; ( S ) Correlation analysis of serum CIRP and IL-1β; ( T ) Correlation analysis of serum CIRP and serum IL-18 in CP patients; ( U ) Correlation analysis of serum CIRP and serum IL-1β in CP patients; ( V ) Correlation analysis of serum CIRP and serum HMGB1 in CP patients. Data are presented as mean ± SD ( n = 6 per group). Statistical analyses were performed using GraphPad Prism 8.0. Statistical significance was determined by t-test or one-way ANOVA with Tukey’s post-hoc test. Correlations were assessed using Spearman’s rank correlation coefficient. * P < 0.05 indicates a statistically significant difference. CP, chronic pancreatitis; CIRP, Cold-inducible RNA-binding protein; L-arg, L-arginine; Cer, Cerulein; IL-6, interleukin 6; TNF-α, tumor necrosis factor-α; NLRP3, Nod-like receptor family protein 3; IL-18, interleukin 18; IL-1β, HMGB1, high mobility group protein B1; interleukin 1β

Article Snippet: In the other groups of experimental CP, normal saline (vehicle) or 5, 20 or 100 mg/kg NLRP3 inhibitor [ ] (S3680, Selleck, Inc. CN) was administered by intraperitoneal injection 2 h after the last injection of L-arginine or cerulein.

Techniques: Injection, Staining, Western Blot, Expressing, RNA Binding Assay

Journal: Inflammation

Article Title: Targeting Cold-Inducible RNA-Binding Protein Attenuates Pancreatic Fibrosis by Suppressing Pyroptosis in Chronic Pancreatitis

doi: 10.1007/s10753-026-02490-x

Figure Lengend Snippet: CIRP deficiency alleviates inflammation and pyroptosis in CP model mice. Cerulein-CP was induced by 6 IP injections of cerulein (50 µg/kg/body weight) twice a week for 10 weeks. L-arginine-CP was induced by 2 hourly IP injections of 4.0 g/kg L-arginine twice a week for 10 weeks. 8 mg/kg C23 was administered 2 h after the last L-arginine or cerulein injection each time. To determine the role of TLR4 in CIRP’s effect in CP, TAK-242, a specific TLR4 receptor inhibitor, were administered through intraperitoneal injection at 1 h after the last injection of L-arginine or cerulein each time. The animals were sacrificed 10 weeks after the first injection of L-arginine or cerulein. Blood and tissue samples were collected. ( A & B ) Western blot analysis and quantitative of the expression of NLRP3, Cleaved Caspase-1 and Cleaved Gasdermin D in the pancreas; ( C-E ) Representative photos and quantitative of F4/80 and CD11b staining; ( F ) Serum IL-1β level; ( G ) Serum IL-18 level; ( H-J ) Western blot analysis and quantitative of the expression of NLRP3 and Cleaved Gasdermin D in the pancreas; ( K & L ) Representative photos and quantitative of F4/80 staining. Data are presented as mean ± SD ( n = 6 per group). Statistical analyses were performed using GraphPad Prism 8.0. Data were analyzed by one-way ANOVA with Tukey’s post-hoc test. * P < 0.05 indicates a statistically significant difference. CP, chronic pancreatitis; CIRP, Cold-inducible RNA-binding protein; KO, knockout; L-arg, L-arginine; NLRP3, Nod-like receptor family protein 3; TLR4, Toll-like receptor 4; IL-18, interleukin 18; IL-1β, interleukin 1β; Cer, Cerulein

Article Snippet: In the other groups of experimental CP, normal saline (vehicle) or 5, 20 or 100 mg/kg NLRP3 inhibitor [ ] (S3680, Selleck, Inc. CN) was administered by intraperitoneal injection 2 h after the last injection of L-arginine or cerulein.

Techniques: Injection, Western Blot, Expressing, Staining, RNA Binding Assay, Knock-Out

Journal: Inflammation

Article Title: Targeting Cold-Inducible RNA-Binding Protein Attenuates Pancreatic Fibrosis by Suppressing Pyroptosis in Chronic Pancreatitis

doi: 10.1007/s10753-026-02490-x

Figure Lengend Snippet: NLRP3 inhibitor alleviates inflammation and pancreatic fibrosis in Experimental CP. Cerulein-CP was induced by 6 IP injections of cerulein (50 µg/kg/body weight) twice a week for 10 weeks. L-arginine-CP was induced by 2 hourly IP injections of 4.0 g/kg L-arginine twice a week for 10 weeks. At 2 h after the last injection of L-arginine or cerulein each time, normal saline (vehicle) or 5, 20, 100 mg/kg NLRP3 inhibitor was administered by intraperitoneal injection. The animals were sacrificed 10 weeks after the first injection of L-arginine or cerulein. Blood and tissue samples were collected. ( A ) Western blot analysis of the expression of CIRP, NLRP3, Cleaved Caspase-1 and Cleaved Gasdermin D in the pancreas; ( B ) Serum IL-1β level; ( C ) Serum IL-6 level; ( D-G ) Representative photos and quantitative of H&E, Sirius red and F4/80 staining; ( H ) Serum HMGB1 level. Data are presented as mean ± SD ( n = 6 per group). Statistical analyses were performed using GraphPad Prism 8.0. Data were analyzed by one-way ANOVA with Tukey’s post-hoc test. * P < 0.05 indicates a statistically significant difference. CP, chronic pancreatitis; CIRP, Cold-inducible RNA-binding protein; NLRP3, Nod-like receptor family protein 3; IL-6, interleukin 6; IL-1β, interleukin 1β; HMGB1, high mobility group protein B1; Cer, Cerulein; L-arg, L-arginine

Article Snippet: In the other groups of experimental CP, normal saline (vehicle) or 5, 20 or 100 mg/kg NLRP3 inhibitor [ ] (S3680, Selleck, Inc. CN) was administered by intraperitoneal injection 2 h after the last injection of L-arginine or cerulein.

Techniques: Injection, Saline, Western Blot, Expressing, Staining, RNA Binding Assay

Journal: Inflammation

Article Title: Targeting Cold-Inducible RNA-Binding Protein Attenuates Pancreatic Fibrosis by Suppressing Pyroptosis in Chronic Pancreatitis

doi: 10.1007/s10753-026-02490-x

Figure Lengend Snippet: Gasdermin D inhibitor alleviates inflammation and pancreatic fibrosis in Experimental CP. Cerulein-CP was induced by 6 IP injections of cerulein (50 µg/kg/body weight) twice a week for 10 weeks. L-arginine-CP was induced by 2 hourly IP injections of 4.0 g/kg L-arginine twice a week for 10 weeks. At 2 h after the last injection of L-arginine or cerulein each time, normal saline (vehicle) or 5, 20, 50 mg/kg Disulfiram, a potent gasdermin D inhibitor, was administered by intraperitoneal injection. The animals were sacrificed 10 weeks after the first injection of L-arginine or cerulein. Blood and tissue samples were collected. ( A-D ) Representative photos and quantitative of H&E, Sirius red and F4/80 staining; ( E ) Western blot analysis of the expression of CIRP, NLRP3 and Cleaved Gasdermin D in the pancreas; ( F ) Serum IL-18 level. Data are presented as mean ± SD ( n = 6 per group). Statistical analyses were performed using GraphPad Prism 8.0. Data were analyzed by one-way ANOVA with Tukey’s post-hoc test. * P < 0.05 indicates a statistically significant difference. CP, chronic pancreatitis; CIRP, Cold-inducible RNA-binding protein; NLRP3, Nod-like receptor family protein 3; IL-18, interleukin 18

Article Snippet: In the other groups of experimental CP, normal saline (vehicle) or 5, 20 or 100 mg/kg NLRP3 inhibitor [ ] (S3680, Selleck, Inc. CN) was administered by intraperitoneal injection 2 h after the last injection of L-arginine or cerulein.

Techniques: Injection, Saline, Staining, Western Blot, Expressing, RNA Binding Assay

Journal: Molecular immunology

Article Title: Complement’s hidden arsenal: new insights and novel functions inside the cell

doi: 10.1016/j.molimm.2017.01.004

Figure Lengend Snippet: Traditional extracellular roles of the complement cascades. The three pathways of complement activation are shown. Although each is triggered independently, yet they all merge at the step of C3 activation. The classical pathway (CP) is initiated by the binding of antibody to antigen and the lectin pathway (LP) by the binding of lectin to an oligosaccaride. The alternative pathway (AP) turns over continuously, generating small amounts of C3(H20) and C3. The AP rapidly amplifies in the presence of pathogens or injured tissue. C3(H20) behaves like C3b in that it can bind FB to initiate the AP. Activation of the complement system leads to inflammation (release of anaphylatoxins C3a and C5a), opsonization (the coating of targets with C3b and/or C4b), and membrane perturbation (formation of the membrane attack complex, MAC, C5b-C9).

Article Snippet: Other intracellular players including the C3a receptor (C3aR), the C5a receptor (C5aR), and factor B (FB) also are currently being evaluated ( Hess and Kemper, 2016 ).

Techniques: Activation Assay, Binding Assay, Membrane

Journal: Molecular immunology

Article Title: Complement’s hidden arsenal: new insights and novel functions inside the cell

doi: 10.1016/j.molimm.2017.01.004

Figure Lengend Snippet: Emerging roles of an intracellular complement system. This system has multiple mechanisms for cell defense, survival and homeostasis, most of which have been demonstrated to operate in many cell types. (A) Intracellular cleavage of C3 stores by a protease generates C3a and C3b (Kolev, Dimeloe, et al., 2015; Liszewski, Kolev, et al., 2013). In CD4+ T cells, this process is essential for induction of T cell effector function. Upon activation, the generated C3b regulates the necessary increase in metabolic processes through mTOR required for a Th1 response, while the C3a, together with the C3aR, are translocated to the cell surface. Additionally, tonic C3a generation is necessary for homeostatic T cell survival. (B) Extracellular opsonization of viruses and bacteria allows for, upon uptake by Ig and complement receptors, intracellular sensing and subsequent activation of mitochondrial antiviral signaling (MAVS) and proteasome mediated viral degradation (Tam, Bidgood, et al., 2014). (C) C3(H2O) is continuously taken up from blood and a majority returned to the cell exterior under steady-state conditions (Elvington, 2016). The C3(H2O) stores retained are a continuous source of intracellular C3a and other fragments. (D) Opsonization of apoptotic cells mediates their intracellular trafficking, delays fusion with the lysosome and regulates presentation of apoptotic cell-associated antigens on MHC II (Baudino, Sardini, et al., 2014). FH also associates with nucleosomes in apoptotic cells, resulting in an anti-inflammatory response (Martin, Leffler, et al., 2016). Not shown is that FH internalized by apoptotic cell facilitates enhanced cleavage of intracellular C3 and thereby apoptotic cell opsonization. (E) Release of intracellular stores of C3 and properdin (P) may be a mechanism to rapidly initiate local complement activation at the site of infection or injury (cell membrane or interstitial space) (Kouser, Abdul-Aziz, et al., 2013; Spitzer, Mitchell, et al., 2007). These five pathways likely interact. C3a and C3b, generated by each, may engage one or more signaling pathways involving Th1/Th17, mTOR, MAVS and nuclear machinery. P, properdin; FH, factor H; FI, factor I; MAVS, mitochondrial antiviral signaling.

Article Snippet: Other intracellular players including the C3a receptor (C3aR), the C5a receptor (C5aR), and factor B (FB) also are currently being evaluated ( Hess and Kemper, 2016 ).

Techniques: Activation Assay, Generated, Bacteria, Infection, Membrane, Protein-Protein interactions