Journal: BMC Nephrology

Article Title: Celastrol modulates damage of renal tissues in Immunoglobulin A nephropathy via targeting TGase-2/HMGB1 signaling pathway

doi: 10.1186/s12882-025-04139-7

Figure Lengend Snippet: RT-PCR primer sequences

Article Snippet: TGase-2 antibody, HMGB1 antibody, IgA antibody (Lot: ab2386, ab79823, ab102198, Abcam Company, UK); TLR4 antibody, MYD88 antibody (Lot:19811-1-AP, 67969-1-Ig, Proteintech, Wuhan, China); HMGB1, IL-6, tumor necrosis factor-α (TNF-α) and FN ELISA kits (Lot: MM-13713H1, MM-0049H2, MM-0122H2, MM-0043H2, respectively; China Jiangsu Enzyme Immunoassay Biotechnology Co., LTD.); DEME medium and Fetal bovine serum (Gibco, Thermo Fisher Scientfic), BCA protein concentration determination kit (enhanced) (Beyotime, Shanghai, China).

Techniques:

Journal: BMC Nephrology

Article Title: Celastrol modulates damage of renal tissues in Immunoglobulin A nephropathy via targeting TGase-2/HMGB1 signaling pathway

doi: 10.1186/s12882-025-04139-7

Figure Lengend Snippet: Celastrol decreased the TGase-2, HMGB1, TLR4 and MYD88 levels in the renal tissue of IgAN rats. ( A ) The protein levels of TGase-2, HMGB1, TLR4 and MYD8 in the renal tissue of IgAN rats were detected by western blots and quantization. ( B ) The mRNA levels of TGase-2, HMGB1, TLR4 and MYD8 in the renal tissue of IgAN rats were detected by RT-qPCR. C, control group; M, model group; CL-H, celastrol high dose group; CL-L, celastrol low dose group; MP, methylprednisolone group. Data are expressed as the means ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001 vs.the control group; ### p < 0.001, ## p < 0.01 vs. the model group

Article Snippet: TGase-2 antibody, HMGB1 antibody, IgA antibody (Lot: ab2386, ab79823, ab102198, Abcam Company, UK); TLR4 antibody, MYD88 antibody (Lot:19811-1-AP, 67969-1-Ig, Proteintech, Wuhan, China); HMGB1, IL-6, tumor necrosis factor-α (TNF-α) and FN ELISA kits (Lot: MM-13713H1, MM-0049H2, MM-0122H2, MM-0043H2, respectively; China Jiangsu Enzyme Immunoassay Biotechnology Co., LTD.); DEME medium and Fetal bovine serum (Gibco, Thermo Fisher Scientfic), BCA protein concentration determination kit (enhanced) (Beyotime, Shanghai, China).

Techniques: Western Blot, Quantitative RT-PCR, Control

Journal: BMC Nephrology

Article Title: Celastrol modulates damage of renal tissues in Immunoglobulin A nephropathy via targeting TGase-2/HMGB1 signaling pathway

doi: 10.1186/s12882-025-04139-7

Figure Lengend Snippet: Celastrol treatment and TGASe-2 knockdown decreased the TGase-2, HMGB1, TLR4 and MYD88 levels in the aIgA1 stimulated HMCs. ( A ) The protein expressions of TGase-2, HMGB1, TLR4 and MYD8 in the aIgA1 stimulated HMCs were detected by western blots and quantization after Celastrol treatment and TGASe-2 knockdown. ( B ) The protein expressions of TGase-2, HMGB1, TLR4 and MYD8 in the aIgA1 stimulated HMCs were detected by RT-qPCR after Celastrol treatment and TGASe-2 knockdown. Data are expressed as the means ± SD. * P < 0.05, ** P < 0.01 vs. the control group; @ P < 0.05, @@ P < 0.01, @@@ P < 0.001 vs. the model group; # P < 0.05, ## P < 0.01, ### P < 0.001 vs. cells transfected with NC-siRNA

Article Snippet: TGase-2 antibody, HMGB1 antibody, IgA antibody (Lot: ab2386, ab79823, ab102198, Abcam Company, UK); TLR4 antibody, MYD88 antibody (Lot:19811-1-AP, 67969-1-Ig, Proteintech, Wuhan, China); HMGB1, IL-6, tumor necrosis factor-α (TNF-α) and FN ELISA kits (Lot: MM-13713H1, MM-0049H2, MM-0122H2, MM-0043H2, respectively; China Jiangsu Enzyme Immunoassay Biotechnology Co., LTD.); DEME medium and Fetal bovine serum (Gibco, Thermo Fisher Scientfic), BCA protein concentration determination kit (enhanced) (Beyotime, Shanghai, China).

Techniques: Knockdown, Western Blot, Quantitative RT-PCR, Control, Transfection

Journal: BMC Nephrology

Article Title: Celastrol modulates damage of renal tissues in Immunoglobulin A nephropathy via targeting TGase-2/HMGB1 signaling pathway

doi: 10.1186/s12882-025-04139-7

Figure Lengend Snippet: RT-PCR primer sequences

Article Snippet: TGase-2 antibody, HMGB1 antibody, IgA antibody (Lot: ab2386, ab79823, ab102198, Abcam Company, UK); TLR4 antibody, MYD88 antibody (Lot:19811-1-AP, 67969-1-Ig, Proteintech, Wuhan, China); HMGB1, IL-6, tumor necrosis factor-α (TNF-α) and FN ELISA kits (Lot: MM-13713H1, MM-0049H2, MM-0122H2, MM-0043H2, respectively; China Jiangsu Enzyme Immunoassay Biotechnology Co., LTD.); DEME medium and Fetal bovine serum (Gibco, Thermo Fisher Scientfic), BCA protein concentration determination kit (enhanced) (Beyotime, Shanghai, China).

Techniques:

Journal: BMC Nephrology

Article Title: Celastrol modulates damage of renal tissues in Immunoglobulin A nephropathy via targeting TGase-2/HMGB1 signaling pathway

doi: 10.1186/s12882-025-04139-7

Figure Lengend Snippet: Celastrol decreased the TGase-2, HMGB1, TLR4 and MYD88 levels in the renal tissue of IgAN rats. ( A ) The protein levels of TGase-2, HMGB1, TLR4 and MYD8 in the renal tissue of IgAN rats were detected by western blots and quantization. ( B ) The mRNA levels of TGase-2, HMGB1, TLR4 and MYD8 in the renal tissue of IgAN rats were detected by RT-qPCR. C, control group; M, model group; CL-H, celastrol high dose group; CL-L, celastrol low dose group; MP, methylprednisolone group. Data are expressed as the means ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001 vs.the control group; ### p < 0.001, ## p < 0.01 vs. the model group

Article Snippet: TGase-2 antibody, HMGB1 antibody, IgA antibody (Lot: ab2386, ab79823, ab102198, Abcam Company, UK); TLR4 antibody, MYD88 antibody (Lot:19811-1-AP, 67969-1-Ig, Proteintech, Wuhan, China); HMGB1, IL-6, tumor necrosis factor-α (TNF-α) and FN ELISA kits (Lot: MM-13713H1, MM-0049H2, MM-0122H2, MM-0043H2, respectively; China Jiangsu Enzyme Immunoassay Biotechnology Co., LTD.); DEME medium and Fetal bovine serum (Gibco, Thermo Fisher Scientfic), BCA protein concentration determination kit (enhanced) (Beyotime, Shanghai, China).

Techniques: Western Blot, Quantitative RT-PCR, Control

Journal: BMC Nephrology

Article Title: Celastrol modulates damage of renal tissues in Immunoglobulin A nephropathy via targeting TGase-2/HMGB1 signaling pathway

doi: 10.1186/s12882-025-04139-7

Figure Lengend Snippet: Celastrol treatment and TGASe-2 knockdown decreased the TGase-2, HMGB1, TLR4 and MYD88 levels in the aIgA1 stimulated HMCs. ( A ) The protein expressions of TGase-2, HMGB1, TLR4 and MYD8 in the aIgA1 stimulated HMCs were detected by western blots and quantization after Celastrol treatment and TGASe-2 knockdown. ( B ) The protein expressions of TGase-2, HMGB1, TLR4 and MYD8 in the aIgA1 stimulated HMCs were detected by RT-qPCR after Celastrol treatment and TGASe-2 knockdown. Data are expressed as the means ± SD. * P < 0.05, ** P < 0.01 vs. the control group; @ P < 0.05, @@ P < 0.01, @@@ P < 0.001 vs. the model group; # P < 0.05, ## P < 0.01, ### P < 0.001 vs. cells transfected with NC-siRNA

Article Snippet: TGase-2 antibody, HMGB1 antibody, IgA antibody (Lot: ab2386, ab79823, ab102198, Abcam Company, UK); TLR4 antibody, MYD88 antibody (Lot:19811-1-AP, 67969-1-Ig, Proteintech, Wuhan, China); HMGB1, IL-6, tumor necrosis factor-α (TNF-α) and FN ELISA kits (Lot: MM-13713H1, MM-0049H2, MM-0122H2, MM-0043H2, respectively; China Jiangsu Enzyme Immunoassay Biotechnology Co., LTD.); DEME medium and Fetal bovine serum (Gibco, Thermo Fisher Scientfic), BCA protein concentration determination kit (enhanced) (Beyotime, Shanghai, China).

Techniques: Knockdown, Western Blot, Quantitative RT-PCR, Control, Transfection

Journal: Molecular Medicine

Article Title: Evobrutinib mitigates neuroinflammation after ischemic stroke by targeting M1 microglial polarization via the TLR4/Myd88/NF-κB pathway

doi: 10.1186/s10020-025-01203-8

Figure Lengend Snippet: Effect of Evobrutinib treatment on microglial subtypes following ischemic stroke. A Flow cytometry analysis of the proportion of M1 microglia in the brain post-stroke. Statistical analysis shows that Evobrutinib treatment reduced the proportion of M1 microglial subtype. B Flow cytometry analysis of the proportion of M2-type microglia in the brain post-stroke. Statistical analysis shows that Evobrutinib treatment increased the proportion of M2 microglial subtype. C qPCR analysis of mRNA expressions of CD86 and iNOS in brain tissues across all groups. D qPCR analysis of mRNA expressions of CD206 and Arg1 in brain tissues across all groups. E Western Blot analysis of TLR4, NF-κB, and MyD88 in the brain post stroke. F – H Statistical analysis of TLR4 ( F ), MyD88 ( G ), and NF-κB ( H ) protein expressions. Mean ± SEM, n = 5 or 6, one-way ANOVA, vs sham ns: no significance; ** P < 0.01, *** P < 0.001; MCAO vs Evobrutinib ns: no significance, ## P < 0.01, ### P < 0.001

Article Snippet: The membranes were blocked with 5% non-fat milk at room temperature for 2 h and then incubated overnight at 4 °C with primary antibodies: Rabbit anti-BTK (1:800, ab208937, abcam), Rabbit anti-BTK phospho Y551 (1:800, ab40770, abcam), Mouse anti-TLR4 (1:800, 66350-1, proteintech), Rabbit anti-Myd88 (1:100, A0980, ABclonal), Rabbit anti-NF-κB (1:800, A2547, ABclonal), Mouse anti-GAPDH (1:1000, KTD101, Abbkine), and Mouse anti-β-actin (1:1000, KTD101, Abbkine).

Techniques: Flow Cytometry, Western Blot

Journal: Molecular Medicine

Article Title: Evobrutinib mitigates neuroinflammation after ischemic stroke by targeting M1 microglial polarization via the TLR4/Myd88/NF-κB pathway

doi: 10.1186/s10020-025-01203-8

Figure Lengend Snippet: Evobrutinib inhibits M1 microglial polarization via the TLR4/MyD88/NF-κB pathway. A Western Blot analysis of BTK and pBTK protein expression levels in microglia from different groups after OGD. B Western Blot analysis of TLR4, NF-κB, and MyD88 expression levels in microglia from different groups after OGD. C Flow cytometry analysis of the effect of Evobrutinib and TAK242 on M1 microglial polarization. D Flow cytometry analysis of the effect of Evobrutinib and TAK242 on M2 microglial polarization. E , F ELISA detection of pro-inflammatory cytokines TNF-α, IL-1β ( E ) and anti-inflammatory cytokines TGF-β, IL-4 ( F ) levels in the supernatant of microglia after OGD and Evobrutinib treatment. Mean ± SEM, n = 4, 5 or 6, one-way ANOVA, vs sham, ns: no significance, * P < 0.05, *** P < 0.001; MCAO vs Evobrutinib ns: no significance, # P < 0.05, ### P < 0.001

Article Snippet: The membranes were blocked with 5% non-fat milk at room temperature for 2 h and then incubated overnight at 4 °C with primary antibodies: Rabbit anti-BTK (1:800, ab208937, abcam), Rabbit anti-BTK phospho Y551 (1:800, ab40770, abcam), Mouse anti-TLR4 (1:800, 66350-1, proteintech), Rabbit anti-Myd88 (1:100, A0980, ABclonal), Rabbit anti-NF-κB (1:800, A2547, ABclonal), Mouse anti-GAPDH (1:1000, KTD101, Abbkine), and Mouse anti-β-actin (1:1000, KTD101, Abbkine).

Techniques: Western Blot, Expressing, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: Stem Cell Research & Therapy

Article Title: Effects of 3D-printed exosome-functionalized brain acellular matrix hydrogel on neuroinflammation in rats following cerebral hemorrhage

doi: 10.1186/s13287-025-04332-3

Figure Lengend Snippet: dECM@exo inhibits ICH-induced inflammation and improves MMP in astrocytes. ( A ) Morphological characteristics of primary astrocytes and their specific marker GFAP. ( B ) dECM@exo transfected into astrocytes. (C, D) Quantitative detection of the proportion of cells expressing Ki67( n = 3). ( E ) Cell viability of cells cultured with dECM or dECM@exo. ( F ) Cell viability of cells cultured with Hemin. ( G - I ) ELISA analysis of IL-1β, TNF-α, and IL-10 levels in astrocyte supernatants. ( J ) Western blot of TLR4 and NF-κB/P65, p-P65. ( K ) Statistical analysis of TLR4. ( L ) Statistical analysis of TLR4 mRNA. Scale bars: A and B, 50 μm; C, 100 μm. Data were expressed as mean ± standard deviation. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: Cells from SHAM, ICH, dECM, and dECM@exo groups were fixed in 4% PFA, blocked with 5% BSA (Sigma, A7906), and incubated overnight at 4 °C with primary antibodies: GFAP (1:200; Abcam, ab7260), TLR4 (1:200; Abcam, ab13556), and NF-κB/P65 (1:200; Abcam, ab307840).

Techniques: Marker, Transfection, Expressing, Cell Culture, Enzyme-linked Immunosorbent Assay, Western Blot, Standard Deviation

Journal: Stem Cell Research & Therapy

Article Title: Effects of 3D-printed exosome-functionalized brain acellular matrix hydrogel on neuroinflammation in rats following cerebral hemorrhage

doi: 10.1186/s13287-025-04332-3

Figure Lengend Snippet: dECM@exo reduces the inflammatory response after ICH. ( A ) Corner test ( n = 12). ( B ) Forelimb placement test ( n = 12). ( C ) Bederson and ( D ) Longa scores were performed from day 1 to day 14 after ICH to evaluate the recovery of neurological function ( n = 12). ( E ) Total leukocyte count in different groups ( n = 12). ( F - H ) ELISA method was used to detect the concentrations of IL-1β, TNF-α and IL-10 in serum ( n = 4). ( I ) Western blot of TLR4 and NF-κB/P65, p-P65. ( J ) Statistical analysis of TLR4 ( n = 3). ( K ) Statistical analysis of TLR4 mRNA ( n = 3). Data were expressed as mean ± standard deviation. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: Cells from SHAM, ICH, dECM, and dECM@exo groups were fixed in 4% PFA, blocked with 5% BSA (Sigma, A7906), and incubated overnight at 4 °C with primary antibodies: GFAP (1:200; Abcam, ab7260), TLR4 (1:200; Abcam, ab13556), and NF-κB/P65 (1:200; Abcam, ab307840).

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Standard Deviation

Journal: Stem Cell Research & Therapy

Article Title: Effects of 3D-printed exosome-functionalized brain acellular matrix hydrogel on neuroinflammation in rats following cerebral hemorrhage

doi: 10.1186/s13287-025-04332-3

Figure Lengend Snippet: dECM@exo inhibits ICH-induced excessive inflammation through the TLR4/NF-κB signaling pathway. ( A ) Western blot of TLR4 and NF-κB/P65, p-P65. ( B ) Statistical analysis of TLR4. ( C ) Statistical analysis of TLR4 mRNA. ( D - F ) ELISA analysis of IL-1β, TNF-α and IL-10 levels in different groups ( n = 3). ( G ) Representative immunofluorescence images and ( I ) statistical analysis of TLR4 expression in astrocytes ( n = 3). ( H ) Representative images and ( J ) statistical analysis of P-65 nuclear translocation in ICH-induced astrocytes ( n = 3). Scale bars: G and H 100 μm. Data were expressed as mean ± standard deviation. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: Cells from SHAM, ICH, dECM, and dECM@exo groups were fixed in 4% PFA, blocked with 5% BSA (Sigma, A7906), and incubated overnight at 4 °C with primary antibodies: GFAP (1:200; Abcam, ab7260), TLR4 (1:200; Abcam, ab13556), and NF-κB/P65 (1:200; Abcam, ab307840).

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Expressing, Translocation Assay, Standard Deviation

Journal: Foods

Article Title: Lactobacillus murinus ZNL-13 Modulates Intestinal Barrier Damage and Gut Microbiota in Cyclophosphamide-Induced Immunosuppressed Mice

doi: 10.3390/foods14081416

Figure Lengend Snippet: Effects of L. murinus ZNL-13 on TLR4/MYD88 pathway in colon of immunosuppressed mice. The protein expression of TLR4, NF-κB, and MyD88 in the colon were detected by Western blot ( a ). The ratios of TLR4/β-actin ( b ), MyD88/β-actin ( c ), and NF-κB/β-actin ( d ) protein bands for each region were quantified using densitometry and presented in the graph. The changes in mRNA levels of IL-1β, IL-2, IL-6, and IL-10 ( e – h ) in colonic tissues, as well as the content of TNF-α, IL-1β, IL-2, IL-6, and IL-10 in colonic tissues, were detected by ELISA ( i – m ). ns p >0.05, * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: After electrophoresis, the membranes were blocked with 5% skim milk powder at 4 °C overnight and then incubated with primary antibodies: β-actin (1:5000, Bioss, Beijing, China), p65 (1:3000, Proteintech, Wuhan, China), TLR4 (1:4000, Proteintech), MYD88 (1:1500, Wanlei, Shenyang, China), Claudin-1 (1:1500, Wanlei, China), Occludin (1:1000, Wanlei, China), ZO-1 (1:1000, Wanlei, China), Bcl-2 (1:1000, Affinity, Liyang, China), Bax (1:4000, Proteintech), caspase-3 (1:1000, Wanlei, China), caspase-8 (1:1000, Wanlei, China), caspase-9 (1:1000, Wanlei, China), and FAS (1:1000, Wanlei, China).

Techniques: Expressing, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Journal of Inflammation Research

Article Title: In vitro Validation of a Novel Disposable Remover to Remove Activated Leukocytes Generated During Cardiopulmonary Bypass: A Pilot Study

doi: 10.2147/JIR.S503575

Figure Lengend Snippet: Antibodies for Flow Cytometric Identification of Activated Cells in Blood

Article Snippet: BV421 , CD284 , BV421 Mouse Anti-Human TLR4 (CD284)(TF901) , BD Pharmingen , 564401.

Techniques: Staining, Blocking Assay

Journal: Pharmaceuticals

Article Title: Kinin B 1 Receptor Agonist Enhances Blood-Brain Barrier Permeability in Healthy and Glioblastoma Environments

doi: 10.3390/ph18040591

Figure Lengend Snippet: B1R and TLR4 labeling patterns in endothelial cells treated with CMT98G and CMU87, with or without DBK. HBMECs cultured in DMEM/F-12 medium (control) ( A , G , M , S ) and treated with DBK ( B , H , N , T ) exhibit diffuse TLR4 ( A ) and B1R ( G ) staining throughout the cell, which becomes concentrated in the perinuclear region upon DBK treatment ( B , H ). HBMECs exposed to CMT98G show reduced TLR4 labeling both in the absence ( C ) and presence ( D ) of DBK, while B1R labeling is prominent in the perinuclear region in cells exposed to CMT98G ( I ) but decreases upon DBK treatment ( J ). In cells exposed to CMU87, TLR4 labeling is reduced without ( E ) and with DBK ( F ), whereas B1R labeling remains intense in the perinuclear region regardless of the presence ( K ) or absence ( L ) of DBK. ( M – R ) Combined images of TLR4, B1R, and DAPI staining. ( S – X ) 3D reconstruction of triple-labeled images. Scale bar: 6 µm Number of experiments = 10.

Article Snippet: The primary antibodies used were rabbit anti-BDKRB1 (dilution 1:300), mouse anti-TLR4 (dilution 1:300), and rabbit anti-ß-actin (ab8227, Abcam, UK) (dilution 1:500).

Techniques: Labeling, Cell Culture, Control, Staining

Journal: Pharmaceuticals

Article Title: Kinin B 1 Receptor Agonist Enhances Blood-Brain Barrier Permeability in Healthy and Glioblastoma Environments

doi: 10.3390/ph18040591

Figure Lengend Snippet: Relative quantification of B1R and TLR4. ( A , B ) In-cell Western assay relative quantification of B1R and TLR4 proteins. B1R protein did not vary under the DBK conditions but decreased significantly in cells exposed to CMT98G and CMU87 without DBK ( A ). TLR4 expression decreased significantly in HBMECs exposed to CMT98G e CMU87 ( B ). ( C , D ) Relative mRNA expression levels of B1R and TLR4 receptors in endothelial cells incubated for 24 h with CMT98G or CMU87, with or without DBK ( C ). B1R mRNA increased in cells exposed to CMT98G and CMU87 compared to the DMEM/F-12 + DBK. CMU87 compared to DMEM/F-12 + DBK ( C ). TLR4 mRNA expression increased in cells treated with CMT98G and CMT98G + DBK compared to the DMEM/F-12 control ( D ). ( A ) * p < 0.05 DMEM/F-12 vs. CMT98G, CMT98G + DBK, CMU87. ( B ) * p < 0.05 DMEM/F-12 vs. CMT98G, CMT98G + DBK, CMU87, CMU87 + DBK. ( C ) * p < 0.05 DMEM/F-12 + DBK vs. CMT98G and CMU87. ( D ) * p < 0.05 DMEM/F-12 vs. CMT98G and CMT98G + DBK (Dunnett’s test and Kruskal–Wallis test). Number of experiments: ( A , B ) 3 and ( C , D ) 9.

Article Snippet: The primary antibodies used were rabbit anti-BDKRB1 (dilution 1:300), mouse anti-TLR4 (dilution 1:300), and rabbit anti-ß-actin (ab8227, Abcam, UK) (dilution 1:500).

Techniques: In-Cell ELISA, Expressing, Incubation, Control

Journal: Pharmaceuticals

Article Title: Kinin B 1 Receptor Agonist Enhances Blood-Brain Barrier Permeability in Healthy and Glioblastoma Environments

doi: 10.3390/ph18040591

Figure Lengend Snippet: Reduction in proximity points between B1R and TLR4 in HBMECs incubated with CMT98G and CMU87 media enriched with DBK. The PLA for observing the proximity between B1R and TLR4 receptors in HBMEC cultures shows binding points under control conditions (DMEM/F-12 medium) ( A , B ), and after 24 h of incubation with CMT98G ( C , D ) and CMU87 ( G , H ), with or without 1 µM DBK ( E , F , I , J ). ( K ) Quantification of binding points reveals a significant reduction in the number of points in the CMU87 + DBK condition compared to DMEM/F-12 and CMT98G. ( K ) * p < 0.05 CMU87 + DBK vs. DMEM/F-12; ** p < 0.05 CMU87 + DBK vs. CMT98G (Tukey’s test). Scale bar: 27 µm. Number of experiments = 3.

Article Snippet: The primary antibodies used were rabbit anti-BDKRB1 (dilution 1:300), mouse anti-TLR4 (dilution 1:300), and rabbit anti-ß-actin (ab8227, Abcam, UK) (dilution 1:500).

Techniques: Incubation, Binding Assay, Control

Journal: Pharmaceuticals

Article Title: Kinin B 1 Receptor Agonist Enhances Blood-Brain Barrier Permeability in Healthy and Glioblastoma Environments

doi: 10.3390/ph18040591

Figure Lengend Snippet: Hypothesis for DBK-mediated transient BBB opening. ( A ) Activation of B1R by DBK promotes the interaction with TLR4, leading to an increase in intracellular calcium concentration. This destabilizes the tight junctions that form the barrier between endothelial cells in blood vessels, resulting in enhanced drug flow into the brain parenchyma. Concurrently, elevated calcium levels stimulate the production of EDHF, which is transported to smooth muscle cells, inducing hyperpolarization. ( B ) DBK-mediated activation of the B1R–TLR4 interaction increases intracellular calcium, disrupting key blood-brain barrier proteins. This disruption may enhance BBB permeability, allowing higher concentrations of chemotherapeutic agents to reach the tumor mass. In addition to DBK, another potential adjuvant at very low doses is LPS, which could induce the differentiation of tumor stem cells, thereby enhancing the efficacy of chemotherapeutic agents, such as TMZ.

Article Snippet: The primary antibodies used were rabbit anti-BDKRB1 (dilution 1:300), mouse anti-TLR4 (dilution 1:300), and rabbit anti-ß-actin (ab8227, Abcam, UK) (dilution 1:500).

Techniques: Activation Assay, Concentration Assay, Disruption, Permeability, Adjuvant