Journal: Journal of Inflammation Research

Article Title: Wheat-Grain Moxibustion Ameliorates Ulcerative Colitis: Suppressing Intestinal Inflammation, Modulating Gut Microbiota, and Restoring Mucosal Barrier Integrity

doi: 10.2147/JIR.S540574

Figure Lengend Snippet: Wgm improves the intestinal mucosal barrier integrity in UC mice. ( A – D ) Serum levels of DAO, D-LA, ICAM-1, and IFABP (n=10). ( E – L ) Immunohistochemical analysis of Claudin 1, MUC2, Occludin, and ZO-1 in colonic tissues (n=4). Scale bars: 100 μm; 50 μm. ( M – P ) Western blot analysis of Claudin 1, Occludin, and ZO-1 expression in colonic tissues (n=3). Data are expressed as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Following the manufacturer’s protocols of ELISA kits, serum levels of ICAM-1 (KE10129, Proteintech), IFABP (E-EL-M0735, Elabscience), DAO (E-EL-M0412, Elabscience), and D-LA (E-BC-K002-M, Elabscience) were measured.

Techniques: Immunohistochemical staining, Western Blot, Expressing

Journal: Cell Proliferation

Article Title: Targeting FABP4 to Inhibit AGEs ‐ RAGE / NF ‐ κB Signalling Effectively Ameliorates Nucleus Pulposus Dysfunction and Angiogenesis in Obesity‐Related Intervertebral Disc Degeneration

doi: 10.1111/cpr.70021

Figure Lengend Snippet: FABP4 played a critical role in mediating obesity‐related IVDD. (A) RT‐qPCR analysis of the expression of FABP4 in NPCs under lipotoxic condition ( n = 3). (B) RT‐qPCR analysis of the expression of FABP4 in IVD tissue from obesity and none‐obesity mice ( n = 3). (C) IHC staining for FABP4 in IVD tissue from obesity and none‐obesity mice. (D) Quantitation result of IHC staining for FABP4 in IVD tissue from obesity and none‐obesity mice ( n = 5). (E) IF staining for the expression of FABP4, COL2A1, and MMP3 in NPCs under lipotoxic condition with or without IL‐1β. (F–H) Quantitation results of the IF staining for the expression of FABP4, COL2A1, and MMP3 in NPCs under lipotoxic condition with or without IL‐1β ( n = 5). (I) Degeneration evaluated by H&E, SOFG, and IHC staining of IVD tissue in WT and FABP4‐KO mice from sham and IVDD group, respectively. (J) Histological score of IVD tissue in WT and FABP4‐KO mice from sham and IVDD group, respectively ( n = 5). (K and L) Quantitation results of IHC staining for ACAN and MMP3 of the IVD tissue in WT and FABP4‐KO mice from sham and IVDD group, respectively ( n = 5). (M) IF staining for COL2A1 in IL‐1β‐induced NPCs with or without silencing FABP4. (N) Quantitation result of IF staining for COL2A1 in IL‐1β‐induced NPCs with or without silencing FABP4 ( n = 5). (O) IF staining for MMP3 in IL‐1β‐induced NPCs with or without silencing FABP4. (P) Quantitation result of IF staining for MMP3 in IL‐1β‐induced NPCs with or without silencing FABP4 ( n = 5). (Q) Degeneration evaluated by H&E, SOFG, and IHC staining of IVD tissue in blank and FABP4‐overexpressed mice from sham and IVDD group, respectively. (R) Histological score of IVD tissue in blank and FABP4‐overexpressed mice from sham and IVDD group, respectively ( n = 5). (S and T) Quantitation results of IHC staining for ACAN and MMP3 of the IVD tissue in blank and FABP4‐overexpressed mice from sham and IVDD group, respectively ( n = 5). (U) IF staining for COL2A1 and MMP3 in IL‐1β‐induced NPCs with or without the administration of recombinant FABP4. (V and W) Quantitation results of the IF staining for COL2A1 and MMP3 in IL‐1β‐induced NPCs with or without the administration of recombinant FABP4 ( n = 5). * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: FABP4 level (E‐EL‐M2404, Elabscience) and VEGF level (E‐EL‐H0111, Elabscience) were measured following the instructions of the ELISA kit.

Techniques: Quantitative RT-PCR, Expressing, Immunohistochemistry, Quantitation Assay, Staining, Recombinant

Journal: Cell Proliferation

Article Title: Targeting FABP4 to Inhibit AGEs ‐ RAGE / NF ‐ κB Signalling Effectively Ameliorates Nucleus Pulposus Dysfunction and Angiogenesis in Obesity‐Related Intervertebral Disc Degeneration

doi: 10.1111/cpr.70021

Figure Lengend Snippet: FABP4 contributed to dysfunction and ECM unbalance in NPCs via activating AGEs/RAGE/NFκB signalling pathway. (A) KEGG analysis of the NPCs with or without the administration of rFABP4 ( n = 3). (B) KEGG analysis of the IVD tissue from WT and KO mice ( n = 5). (C) ELISA result of AGEs level in the supernatant of the NPCs treated by rFABP4 in a dose‐dependent manner ( n = 5). (D) ELISA result of AGEs level in the supernatant of the NPCs under lipotoxic condition with or without the treatment of rFABP4, FPS‐ZM1 (an inhibitor for RAGE), and JSH23 (an inhibitor for p65) ( n = 5). (E) IF staining for COL2A1, MMP3, and p65 in NPCs under lipotoxic condition with or without the treatment of rFABP4, FPS‐ZM1, and JSH23. (F–H) Quantitation results of the IF staining for COL2A1, MMP3, and p65 in NPCs under lipotoxic condition with or without the treatment of rFABP4, FPS‐ZM1, and JSH23 ( n = 5). (I) IF staining for RAGE, p‐p65, COL2A1, and MMP3 in WT and KO mice from sham or IVDD groups under HFD condition, respectively. (J) Quantitation results of the IF staining for RAGE, p‐p65, COL2A1, and MMP3 in WT and KO mice from sham or IVDD groups under HFD condition, respectively ( n = 5). (K) IF staining for COL2A1, MMP3, and p65 in NPCs treated by AGEs with or without the treatment of FPS‐ZM1 and JSH23. (L and M) Quantitation results of the F staining for COL2A1, MMP3, and p65 in NPCs treated by AGEs with or without the treatment of FPS‐ZM1 and JSH23 ( n = 5). * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: FABP4 level (E‐EL‐M2404, Elabscience) and VEGF level (E‐EL‐H0111, Elabscience) were measured following the instructions of the ELISA kit.

Techniques: Enzyme-linked Immunosorbent Assay, Staining, Quantitation Assay

Journal: Cell Proliferation

Article Title: Targeting FABP4 to Inhibit AGEs ‐ RAGE / NF ‐ κB Signalling Effectively Ameliorates Nucleus Pulposus Dysfunction and Angiogenesis in Obesity‐Related Intervertebral Disc Degeneration

doi: 10.1111/cpr.70021

Figure Lengend Snippet: Obesity‐induced expression of FABP4 was regulated by the mTORC1 pathway. (A) RT‐qPCR for the expression of FABP4, ACAN, COL2A1, MMP3, and MMP13 in NPCs under lipotoxic condition treated with or without rapamycin (a specific inhibitor of the mTOR pathway) ( n = 3). (B) ELISA result of FABP4 and AGEs levels under lipotoxic condition treated with or without rapamycin ( n = 5). (C–F) IF staining for FABP4, COL2A1, MMP3, PS6, and p65 in NPCs under lipotoxic condition treated with or without rapamycin. (G) Quantitation results of the IF staining for FABP4, COL2A1, MMP3, and PS6 in NPCs under lipotoxic condition treated with or without rapamycin ( n = 5). (H) Quantitation results of the IF staining for nucleus translocation of p65 in NPCs under lipotoxic condition treated with or without rapamycin ( n = 5). (I) Degeneration and mTORC1 pathway activation evaluated by H&E, SOFG, and IHC staining of IVD tissue in sham, IVDD, and IVDD+ rapamycin groups, respectively. (J) Histological score of IVD tissue in sham, IVDD, and IVDD+ rapamycin groups, respectively ( n = 5). (K) Quantitation results of the IHC staining for COL2A1, ACAN, and PS6 in sham, IVDD, and IVDD+ rapamycin groups, respectively ( n = 5). (L and M) IF staining and quantitation results for FABP4 in NPCs treated with or without MHY1485 (an activator for mTOR pathway). N and O: IF staining and quantitation results for COL2A1 and MMP3 in NPCs treated with or without MHY1485 (an activator for mTOR pathway). * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: FABP4 level (E‐EL‐M2404, Elabscience) and VEGF level (E‐EL‐H0111, Elabscience) were measured following the instructions of the ELISA kit.

Techniques: Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Staining, Quantitation Assay, Translocation Assay, Activation Assay, Immunohistochemistry

Journal: Cell Proliferation

Article Title: Targeting FABP4 to Inhibit AGEs ‐ RAGE / NF ‐ κB Signalling Effectively Ameliorates Nucleus Pulposus Dysfunction and Angiogenesis in Obesity‐Related Intervertebral Disc Degeneration

doi: 10.1111/cpr.70021

Figure Lengend Snippet: FABP4 promoted angiogenesis to exacerbate IVDD under lipotoxic condition. (A) Volcano plot of the DEGs of HFD‐induced IVDD from WT and KO mice. (B) Heart map of the DEGs of HFD‐induced IVDD from WT and KO mic ( n = 5). (C) GO analysis of the DEGs of HFD‐induced IVDD from WT and KO mice. (D) KEGG analysis of the DEGs of HFD‐induced IVDD from WT and KO mice. (E) ELISA result of the VEGF level of the IVD tissue in WT and KO mice ( n = 5). (F) ELISA result of the VEGF level of NPC degeneration model with or without FABP4 silencing or FPS‐ZM1 ( n = 5). (G) IF staining for CD31 and EMCN in WT and KO mice from sham or IVDD groups under HFD condition, respectively. (H and I) Quantitation results of the IF staining for CD31 and EMCN in WT and KO mice from sham or IVDD groups under HFD condition, respectively ( n = 5). (J) Tube formation assay of HUVECs in different groups. (K and L) Branch points and capillary length of HUVECs in different groups ( n = 5). (M and N) Representative images and quantitation results of the scratch wound of HUVECs in different groups ( n = 5). * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: FABP4 level (E‐EL‐M2404, Elabscience) and VEGF level (E‐EL‐H0111, Elabscience) were measured following the instructions of the ELISA kit.

Techniques: Enzyme-linked Immunosorbent Assay, Staining, Quantitation Assay, Tube Formation Assay

Journal: Cell Proliferation

Article Title: Targeting FABP4 to Inhibit AGEs ‐ RAGE / NF ‐ κB Signalling Effectively Ameliorates Nucleus Pulposus Dysfunction and Angiogenesis in Obesity‐Related Intervertebral Disc Degeneration

doi: 10.1111/cpr.70021

Figure Lengend Snippet: Blocking the interaction of VEGF and VEGFR2 alleviated FABP4‐induced angiogenesis and IVDD (A) Degeneration and angiogenesis evaluated by H&E, SOFG, and IHC staining of IVD tissue from sham, IVDD, IVDD+rFABP4, and IVDD+rFABP4 + Ki8751 groups, respectively. (B) Histological score of IVD tissue from sham, IVDD, IVDD+rFABP4, and IVDD+rFABP4 + Ki8751 groups, respectively ( n = 5). (C and D): Quantitation results of IF staining for CD31 and EMCN of IVD tissue from sham, IVDD, IVDD+rFABP4, and IVDD+rFABP4 + Ki8751 groups, respectively ( n = 5). (E and F) Quantitation results of IHC staining for COL2A1 and MMP3 of IVD tissue from sham, IVDD, IVDD+rFABP4, and IVDD+rFABP4 + Ki8751 groups, respectively ( n = 5). (G) Tube formation assay of HUVECs in different groups. (H and I) Branch points and capillary length of HUVECs in different groups ( n = 5). (J and K) Representative images and quantitation results of the scratch wound of HUVECs in different groups ( n = 5). * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: FABP4 level (E‐EL‐M2404, Elabscience) and VEGF level (E‐EL‐H0111, Elabscience) were measured following the instructions of the ELISA kit.

Techniques: Blocking Assay, Immunohistochemistry, Quantitation Assay, Staining, Tube Formation Assay

Journal: Cell Proliferation

Article Title: Targeting FABP4 to Inhibit AGEs ‐ RAGE / NF ‐ κB Signalling Effectively Ameliorates Nucleus Pulposus Dysfunction and Angiogenesis in Obesity‐Related Intervertebral Disc Degeneration

doi: 10.1111/cpr.70021

Figure Lengend Snippet: Inhibition of FABP4 alleviated obesity‐related IVDD via restoring ECM balance and inhibiting angiogenesis (A) Representative MRI images of IVD in sham, IVDD, and IVDD+BMS groups, respectively. (B) Degeneration evaluated by H&E, SOFG, and IHC staining of IVD tissue from sham, IVDD, and IVDD+BMS groups, respectively. (C–H) Histological score of IVD from sham, IVDD, and IVDD+BMS groups, respectively ( n = 5). (I) Quantitation results of IHC staining for COL2A1, ACAN, and MMP3 of IVD from sham, IVDD, and IVDD+BMS groups, respectively ( n = 5). (J–L) IF staining for COL2A1, MMP3, CD31, and EMCN of IVD from sham, IVDD, and IVDD+BMS groups, respectively. (M) Quantitation results of IF staining for COL2A1, MMP3, CD31, and EMCN of IVD from sham, IVDD, and IVDD+BMS groups, respectively ( n = 5).

Article Snippet: FABP4 level (E‐EL‐M2404, Elabscience) and VEGF level (E‐EL‐H0111, Elabscience) were measured following the instructions of the ELISA kit.

Techniques: Inhibition, Immunohistochemistry, Quantitation Assay, Staining

Journal: Nutrients

Article Title: Tryptophan Attenuates Chronic Restraint Stress-Induced Intestinal Injury Through Modulation of Intestinal Barrier Integrity and Gut Microbiota Homeostasis

doi: 10.3390/nu17060975

Figure Lengend Snippet: Figure 3. Tryptophan alleviates CRS-induced intestine leakage and epithelial apoptosis. (A) Serum intestinal fatty acid-binding protein content by ELISA; (B) FITC-dextran content in serum; (C) TUNEL staining plots; (D) TUNEL-positive cell counts; different letters denote significant differences between groups, p < 0.05. Ctrl refers to control group.

Article Snippet: We obtained the Mouse Corticosterone (CORT) ELISA Kit (Elabscience, Shanghai, China), FITC—dextran (4 kD) (Sigma, Saint Louis, MI, USA); the mouse intestinal fatty acid-binding protein (FABP) ELISA Kit (CUSABIO, Wuhan, China); and all of the 11 SCFA standards (ZZ Standards Co., Ltd., Shanghai, China).

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, TUNEL Assay, Staining, Control