Journal: iScience

Article Title: Zhigancao decoction alleviates Parkinson’s disease via inhibiting TNF/NF-κB and Ras/ERK-mediated neuroinflammation and apoptosis

doi: 10.1016/j.isci.2025.114489

Figure Lengend Snippet: ZGCD treatment may alleviate MPTP-induced injury of SN DA neurons by inhibiting the TNF/NF-kB and Ras/ERK pathways (A) WB was used to detect the representative expressions of TNFR1, P65, and p-P65 in SN. (B and C) Statistics of the relative expression level of TNFR1 and p-P65/P65 in SN ( n = 4). (D) WB was used to detect the representative expressions of Ras, ERK, and p -ERK in SN. (E and F) Statistics of the relative expression level of Ras and p -ERK/ERK in SN ( n = 4). (G–K) Representative expression of Tnfr1, Nfkb, Grb2, Ras, and Erk detected by RT-qPCR ( n = 4). ### p < 0.001 vs. Con; n.s., not significant, ∗ p < 0.05, ∗∗ p < 0.01 and ∗∗∗ p < 0.001 vs. MPTP. Data are expressed as mean ± SD.

Article Snippet: To verify the potential connection between TNF/NF-κB and Ras/ERK, we observed the activation changes of the two pathways and the therapeutic effect of ZGCDS after treatment with the TNFR1 inhibitor R-7050 (MCE, China, HY-110203) in vitro .

Techniques: Expressing, Quantitative RT-PCR

Journal: iScience

Article Title: Zhigancao decoction alleviates Parkinson’s disease via inhibiting TNF/NF-κB and Ras/ERK-mediated neuroinflammation and apoptosis

doi: 10.1016/j.isci.2025.114489

Figure Lengend Snippet: ZGCDS exerts neuroprotective effects by inhibiting the activation of TNF/NF-κB and Ras/ERK signaling pathways in SH-SY5Y cells (A) WB was used to detect the representative expressions of TNFR1, P65, and p-P65 in SH-SY5Y cells. (B) Statistics of the relative expression level of TNFR1 in SH-SY5Y cells ( n = 3). (C) Statistics of the relative expression level of p-P65/P65 in SH-SY5Y cells ( n = 3). (D–G) IF was used to analyze the expressions of TNFR1 and p-P65 in SH-SY5Y cells ( n = 3). Scale bars, 200 μm. (H–J) WB analysis of Ras and ERK expression and phosphorylation in SH-SY5Y cells ( n = 6). (K–P) IF was used to analyze the expressions of Grb2, Ras, and p -ERK in SH-SY5Y cells ( n = 3). Scale bars, 100 μm. (Q–S) WB analysis of TNFR1 and P65 expression and phosphorylation in SH-SY5Y cells ( n = 4). (T–W) WB analysis of Grb2, Ras, and ERK expression and phosphorylation in SH-SY5Y cells ( n = 4). ### p < 0.001 vs. Con; n.s., not significant, ∗ p < 0.05, ∗∗ p < 0.01 and ∗∗∗ p < 0.001 vs. MPTP. Data are expressed as mean ± SD.

Article Snippet: To verify the potential connection between TNF/NF-κB and Ras/ERK, we observed the activation changes of the two pathways and the therapeutic effect of ZGCDS after treatment with the TNFR1 inhibitor R-7050 (MCE, China, HY-110203) in vitro .

Techniques: Activation Assay, Protein-Protein interactions, Expressing, Phospho-proteomics

Journal: iScience

Article Title: Zhigancao decoction alleviates Parkinson’s disease via inhibiting TNF/NF-κB and Ras/ERK-mediated neuroinflammation and apoptosis

doi: 10.1016/j.isci.2025.114489

Figure Lengend Snippet: Bioactive compounds (Apigenin, Bisdemethoxycurcumin) from ZGCD may exert anti-PD effects by targeting the TNF/NF-kB and Ras/ERK pathways (A and B) Total ion chromatograms (TICs) of Con in positive (A) and negative (B) ion modes. (C and D) Total ion chromatograms (TICs) of ZGCD-CS in positive (C) and negative (D) ion modes. (E) Venn diagram of serum pharmacochemical analysis. (F) Molecular docking analysis of Apigenin and Bisdemethoxycurcumin with key proteins in the pathway. (G–I) The levels of TNF-α, IL-1β, and IL-6 were detected by ELISA in SH-SY5Y cells ( n = 3). (J–L) WB analysis of TNFR1 and P65 expression and phosphorylation in SH-SY5Y cells ( n = 3). (M–P) WB analysis of Grb2, Ras, and ERK expression and phosphorylation in SH-SY5Y cells ( n = 3). ### p < 0.001 vs. Con; n.s., not significant, ∗ p < 0.05, ∗∗ p < 0.01 and ∗∗∗ p < 0.001 vs. MPTP. Data are expressed as mean ± SD.

Article Snippet: To verify the potential connection between TNF/NF-κB and Ras/ERK, we observed the activation changes of the two pathways and the therapeutic effect of ZGCDS after treatment with the TNFR1 inhibitor R-7050 (MCE, China, HY-110203) in vitro .

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Phospho-proteomics

Journal: Journal of Inflammation Research

Article Title: Deciphering the Therapeutic Mechanisms of Banxia Xiexin Decoction Against Ulcerative Colitis: Targeting Pyroptosis and Necroptosis

doi: 10.2147/JIR.S564333

Figure Lengend Snippet: BXD reversed pyroptosis and necroptosis via the P2RX7/NEK7 and TNFR1/RIPK3 pathways in DSS mice. ( A ) IHC results of p-MLKL and GSDMD expression. ( B ) Mean OD density of p-MLKL and GSDMD immunostaining (n=4). ( C and D ) Western blot analysis of colonic P2RX7, NEK7, NLRP3, caspase-1, cleaved caspase-1, ASC, GSDMD, GSDMD-N and cleaved IL-1β (n=3). ( E and F ) Western blot analysis of colonic TNFR1, p-RIPK1, RIPK1, p-RIPK3, RIPK3, p-MLKL and MLKL (n=3). Data were presented as mean±SEM. ### P < 0.001, ## P < 0.01, # P < 0.05 vs control group; *** P < 0.001, ** P < 0.01, * P < 0.05 vs DSS group.

Article Snippet: Primary antibodies used included NF-κB p65 (1:2000, GB12142, Servicebio), p-NF-κB p65 (1:1000, 3033S, Cell Signaling technology), ZO-1 (1:2000, GB111402 , Servicebio), P2RX7 (1:5000, 28,207-1-AP, Proteintech), NEK7 (1:10000, ab133514, Abcam), NLRP3 (1:1000, T55651S, Abmart), ASC (1:5000, 10,500-1-AP, Proteintech), caspase-1 (1:1000, ab179515, Abcam), GSDMD (1:5000, ab219800, Abcam), cleaved IL-1β (1:1000, 63124S, Cell Signaling technology), TNFR1 (1:500, 21,574-1-AP, Proteintech), p-RIPK1 (1:500, AP1314, ABclonal), RIPK1 (1:1000, A27859 , ABclonal), p-RIPK3 (1:1000, 91702, Cell Signaling technology), RIPK3 (1:2000, YM8350, Immunoway), p-MLKL (1:1000, YP1884, Immunoway), MLKL (1:2000, YM8455, Immunoway), GADPH (1:50,000, 60,004-1-Ig, Proteintech), and β-actin (1:10000, GB11001, Servicebio), which were incubated overnight at 4°C.

Techniques: Expressing, Immunostaining, Western Blot, Control

Journal: Journal of Inflammation Research

Article Title: Deciphering the Therapeutic Mechanisms of Banxia Xiexin Decoction Against Ulcerative Colitis: Targeting Pyroptosis and Necroptosis

doi: 10.2147/JIR.S564333

Figure Lengend Snippet: BXD inhibited pyroptosis and necroptosis via the P2RX7/NEK7 and TNFR1/RIPK3 pathways in NCM460 cells. ( A and B ) Western blot analysis of P2RX7, NEK7, NLRP3, caspase-1, cleaved caspase-1, ASC, GSDMD, GSDMD-N, and cleaved IL-1β ( C and D ) Western blot analysis of TNFR1, p-RIPK1, RIPK1, p-RIPK3, RIPK3, p-MLKL and MLKL. Data were presented as mean±SEM (n=3). ### P < 0.001, # P < 0.05 vs control group; *** P < 0.001, ** P < 0.01, * P < 0.05 vs LPS+TNF-α group.

Article Snippet: Primary antibodies used included NF-κB p65 (1:2000, GB12142, Servicebio), p-NF-κB p65 (1:1000, 3033S, Cell Signaling technology), ZO-1 (1:2000, GB111402 , Servicebio), P2RX7 (1:5000, 28,207-1-AP, Proteintech), NEK7 (1:10000, ab133514, Abcam), NLRP3 (1:1000, T55651S, Abmart), ASC (1:5000, 10,500-1-AP, Proteintech), caspase-1 (1:1000, ab179515, Abcam), GSDMD (1:5000, ab219800, Abcam), cleaved IL-1β (1:1000, 63124S, Cell Signaling technology), TNFR1 (1:500, 21,574-1-AP, Proteintech), p-RIPK1 (1:500, AP1314, ABclonal), RIPK1 (1:1000, A27859 , ABclonal), p-RIPK3 (1:1000, 91702, Cell Signaling technology), RIPK3 (1:2000, YM8350, Immunoway), p-MLKL (1:1000, YP1884, Immunoway), MLKL (1:2000, YM8455, Immunoway), GADPH (1:50,000, 60,004-1-Ig, Proteintech), and β-actin (1:10000, GB11001, Servicebio), which were incubated overnight at 4°C.

Techniques: Western Blot, Control

Journal: Journal of Inflammation Research

Article Title: Deciphering the Therapeutic Mechanisms of Banxia Xiexin Decoction Against Ulcerative Colitis: Targeting Pyroptosis and Necroptosis

doi: 10.2147/JIR.S564333

Figure Lengend Snippet: Molecular docking and SPR screening of BXD bioactive components targeting P2RX7 and TNFR1. ( A ) Molecular docking analysis of the bioactive components in BXD with P2RX7 and TNFR1 target proteins. The positive controls A839977 and balinatunfib were included for comparison. ( B and C ) SPR screening of the top five BXD compounds that bind to P2RX7 and TNFR1. ( D and E ) SPR analysis of Baicalin binding to P2RX7 and glycyrrhizic acid binding to TNFR1. ( F ) Visual representation of the protein-ligand interaction profiles between baicalin, A839977 and P2RX7, as well as between glycyrrhizic acid, balinatunfib and TNFR1.

Article Snippet: Primary antibodies used included NF-κB p65 (1:2000, GB12142, Servicebio), p-NF-κB p65 (1:1000, 3033S, Cell Signaling technology), ZO-1 (1:2000, GB111402 , Servicebio), P2RX7 (1:5000, 28,207-1-AP, Proteintech), NEK7 (1:10000, ab133514, Abcam), NLRP3 (1:1000, T55651S, Abmart), ASC (1:5000, 10,500-1-AP, Proteintech), caspase-1 (1:1000, ab179515, Abcam), GSDMD (1:5000, ab219800, Abcam), cleaved IL-1β (1:1000, 63124S, Cell Signaling technology), TNFR1 (1:500, 21,574-1-AP, Proteintech), p-RIPK1 (1:500, AP1314, ABclonal), RIPK1 (1:1000, A27859 , ABclonal), p-RIPK3 (1:1000, 91702, Cell Signaling technology), RIPK3 (1:2000, YM8350, Immunoway), p-MLKL (1:1000, YP1884, Immunoway), MLKL (1:2000, YM8455, Immunoway), GADPH (1:50,000, 60,004-1-Ig, Proteintech), and β-actin (1:10000, GB11001, Servicebio), which were incubated overnight at 4°C.

Techniques: Comparison, Binding Assay

Journal: Journal of Inflammation Research

Article Title: Deciphering the Therapeutic Mechanisms of Banxia Xiexin Decoction Against Ulcerative Colitis: Targeting Pyroptosis and Necroptosis

doi: 10.2147/JIR.S564333

Figure Lengend Snippet: Molecular mechanism diagram. Banxia Xiexin decoction reduces pyroptosis and necroptosis by inhibiting the P2RX7/NEK7 and TNFR1/RIPK3 signaling.

Article Snippet: Primary antibodies used included NF-κB p65 (1:2000, GB12142, Servicebio), p-NF-κB p65 (1:1000, 3033S, Cell Signaling technology), ZO-1 (1:2000, GB111402 , Servicebio), P2RX7 (1:5000, 28,207-1-AP, Proteintech), NEK7 (1:10000, ab133514, Abcam), NLRP3 (1:1000, T55651S, Abmart), ASC (1:5000, 10,500-1-AP, Proteintech), caspase-1 (1:1000, ab179515, Abcam), GSDMD (1:5000, ab219800, Abcam), cleaved IL-1β (1:1000, 63124S, Cell Signaling technology), TNFR1 (1:500, 21,574-1-AP, Proteintech), p-RIPK1 (1:500, AP1314, ABclonal), RIPK1 (1:1000, A27859 , ABclonal), p-RIPK3 (1:1000, 91702, Cell Signaling technology), RIPK3 (1:2000, YM8350, Immunoway), p-MLKL (1:1000, YP1884, Immunoway), MLKL (1:2000, YM8455, Immunoway), GADPH (1:50,000, 60,004-1-Ig, Proteintech), and β-actin (1:10000, GB11001, Servicebio), which were incubated overnight at 4°C.

Techniques: