Journal: Functional & Integrative Genomics

Article Title: Huang Qin decoction increases SLC6A4 expression and blocks the NFκB-mediated NLRP3/Caspase1/GSDMD pathway to disrupt colitis-associated carcinogenesis

doi: 10.1007/s10142-024-01334-x

Figure Lengend Snippet: HQD affects SLC6A4 expression and serotonin production in mouse colon tissues. (A) The ingredient-target-pathway/disease network of HQD in BATMAN-TCM. (B) The intersection of therapeutic targets screened in BATMAN-TCM and genes affected by Scutellaria baicalensis , Paeonia lactiflora , Glycyrrhiza uralensis , and Ziziphus jujuba in HERB 2.0. (C) Changes in SLC6A4 mRNA expression in AOM/DSS mice after HQD treatment detected by RT-qPCR. (D) Changes in SLC6A4 protein expression in AOM/DSS mice after HQD treatment detected by western blot assays. (E) Detection of serotonin (5-HT) production in mouse colon tissues by ELISA. The data are presented as the means ± SDs ( n = 6). *p < 0.05, ** p < 0.01, **** p < 0.0001. The data were analyzed by one-way ANOVA and Tukey’s multiple comparison test

Article Snippet: After being blocked with skim milk for 45 min at room temperature, the PVDF membranes were incubated overnight at 4 °C with primary antibodies against SLC6A4 (1:1000, H00006532-D01P, Novus Biological Inc., Littleton, CO, USA), p-NFκB p65 (1:1000, #3033, Cell Signaling Technologies, Beverly, MA, USA), NFκB p65 (1:1000, #8242, Cell Signaling Technologies), NLRP3 (1:1000, #15,101, Cell Signaling Technologies), Cleaved-Caspase-1 P20 (1:1000, PA5-99390, Invitrogen), Cleaved-GSDMD (1:1000, #50,928, Cell Signaling Technologies), GSDMD (1:1000, ab225867, Abcam, Cambridge, UK), Caspase1 (1:1000, ab138483, Abcam), ASC (1:1000, #67,824, Cell Signaling Technologies), and β-actin (1:200, ab115777, Abcam).

Techniques: Expressing, Biomarker Discovery, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Functional & Integrative Genomics

Article Title: Huang Qin decoction increases SLC6A4 expression and blocks the NFκB-mediated NLRP3/Caspase1/GSDMD pathway to disrupt colitis-associated carcinogenesis

doi: 10.1007/s10142-024-01334-x

Figure Lengend Snippet: The therapeutic effect of HQD on AOM/DSS mice is inhibited after SLC6A4 knockout. AOM/DSS was used to establish WT and SLC6A4-deficient mice, which were subsequently treated with HQD. (A) Changes in the body weight of mice. (B) DAI of the mice. (C) Length of colon tissue in mice. (D) Number of tumors in the colon tissues of the mice. (E) HE staining of pathological morphology of the colon tissues. (F) Detection of 5-HT, IL-1β and IL-18 levels in mouse colon tissues by ELISA. (G) The expression of the p-NFκB p65, NLRP3, Cleaved-Caspase1 P20, Cleaved-GSDMD, and ASC proteins was determined in the colon tissue of AOM/DSS mice by western blot assay. The data are presented as the means ± SDs ( n = 6). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. The data were analyzed by an unpaired t-test or two-way ANOVA and Tukey’s multiple comparison test

Article Snippet: After being blocked with skim milk for 45 min at room temperature, the PVDF membranes were incubated overnight at 4 °C with primary antibodies against SLC6A4 (1:1000, H00006532-D01P, Novus Biological Inc., Littleton, CO, USA), p-NFκB p65 (1:1000, #3033, Cell Signaling Technologies, Beverly, MA, USA), NFκB p65 (1:1000, #8242, Cell Signaling Technologies), NLRP3 (1:1000, #15,101, Cell Signaling Technologies), Cleaved-Caspase-1 P20 (1:1000, PA5-99390, Invitrogen), Cleaved-GSDMD (1:1000, #50,928, Cell Signaling Technologies), GSDMD (1:1000, ab225867, Abcam, Cambridge, UK), Caspase1 (1:1000, ab138483, Abcam), ASC (1:1000, #67,824, Cell Signaling Technologies), and β-actin (1:200, ab115777, Abcam).

Techniques: Knock-Out, Staining, Enzyme-linked Immunosorbent Assay, Expressing, Western Blot, Comparison

Journal: Functional & Integrative Genomics

Article Title: Huang Qin decoction increases SLC6A4 expression and blocks the NFκB-mediated NLRP3/Caspase1/GSDMD pathway to disrupt colitis-associated carcinogenesis

doi: 10.1007/s10142-024-01334-x

Figure Lengend Snippet: SLC6A4 downregulation promotes MODE-K cell pyroptosis treated with DS. MODE-K cells were infected with sh-SLC6A4 or sh-NC, pre-treated with DS, and induced with LPS-ATP. (A) Detection of SLC6A4 mRNA expression in MODE-K cells after infection by RT-qPCR. (B) MODE-K cell viability was examined using a CCK-8 assay. (C) Measurement of LDH release. (D) The expression of NLRP3, Cleaved-Caspase1 P20, and Cleaved-GSDMD in MODE-K cells was examined using a western blot. (E) Immunofluorescence detection of ASC expression in MODE-K cells. The data are presented as the means ± SD ( n = 3). ** p < 0.01, **** p < 0.0001. The data were analyzed by unpaired t-test or two-way ANOVA and Tukey’s multiple comparison test

Article Snippet: After being blocked with skim milk for 45 min at room temperature, the PVDF membranes were incubated overnight at 4 °C with primary antibodies against SLC6A4 (1:1000, H00006532-D01P, Novus Biological Inc., Littleton, CO, USA), p-NFκB p65 (1:1000, #3033, Cell Signaling Technologies, Beverly, MA, USA), NFκB p65 (1:1000, #8242, Cell Signaling Technologies), NLRP3 (1:1000, #15,101, Cell Signaling Technologies), Cleaved-Caspase-1 P20 (1:1000, PA5-99390, Invitrogen), Cleaved-GSDMD (1:1000, #50,928, Cell Signaling Technologies), GSDMD (1:1000, ab225867, Abcam, Cambridge, UK), Caspase1 (1:1000, ab138483, Abcam), ASC (1:1000, #67,824, Cell Signaling Technologies), and β-actin (1:200, ab115777, Abcam).

Techniques: Infection, Expressing, Quantitative RT-PCR, CCK-8 Assay, Western Blot, Immunofluorescence, Comparison

Journal: Functional & Integrative Genomics

Article Title: Huang Qin decoction increases SLC6A4 expression and blocks the NFκB-mediated NLRP3/Caspase1/GSDMD pathway to disrupt colitis-associated carcinogenesis

doi: 10.1007/s10142-024-01334-x

Figure Lengend Snippet: A schematic drawing of HQD in CAC. Activation of SLC6A4 expression by HQD decreased the secretion of serotonin. After serotonin secretion was reduced, the NFκB pathway-induced NLRP3/Caspase1/GSDMD pyroptosis pathway was blocked, which alleviated CAC.

Article Snippet: After being blocked with skim milk for 45 min at room temperature, the PVDF membranes were incubated overnight at 4 °C with primary antibodies against SLC6A4 (1:1000, H00006532-D01P, Novus Biological Inc., Littleton, CO, USA), p-NFκB p65 (1:1000, #3033, Cell Signaling Technologies, Beverly, MA, USA), NFκB p65 (1:1000, #8242, Cell Signaling Technologies), NLRP3 (1:1000, #15,101, Cell Signaling Technologies), Cleaved-Caspase-1 P20 (1:1000, PA5-99390, Invitrogen), Cleaved-GSDMD (1:1000, #50,928, Cell Signaling Technologies), GSDMD (1:1000, ab225867, Abcam, Cambridge, UK), Caspase1 (1:1000, ab138483, Abcam), ASC (1:1000, #67,824, Cell Signaling Technologies), and β-actin (1:200, ab115777, Abcam).

Techniques: Activation Assay, Expressing