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Sauchinone inhibits M1 macrophage polarization and promotes M2 macrophage polarization in BMDM. (A) The cytotoxicity of Sauchinone on BMDM was detected by CCK-8 kit. (B-G) BMDM were stimulated with LPS plus IFN-γ to induce M1 polarization in the presence or absence of different doses of Sauchinone. The transcription levels of Tnf-α , Il-6 , Il −12 , Inos , Il-1β , Cox-2 and Nlrp3 (B) were detected by RT-qPCR. TNF-α, IL-6, IL-12 and NO levels in culture supernatants (C) were measured by ELISA or Griess assay. The protein levels of <t>p-STAT-1,</t> STAT-1, and INOS in cell lysates (D) were detected by western blot. The percentages of INOS + , MHCⅡ + and CD86 + BMDM (E) were measured by FACS. (F-H) BMDM were stimulated with IL-4 plus IL-13 to induce M2 polarization in the presence or absence of different doses of Sauchinone. The transcription levels of Arg-1, Fizz1 , Ym-1 , Cd206 , Cd301 , and Dectin-1 (F) were detected by RT-qPCR. The protein levels of p-STAT-6 and STAT-6 in cell lysates (G) were detected by western blot. The percentages of CD301 + BMDM (H) were measured by FACS. Data were shown as Mean ± SEM from triplicate measurements. *P < 0.05, **P < 0.01 compared as indicated.
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Sauchinone inhibits M1 macrophage polarization and promotes M2 macrophage polarization in BMDM. (A) The cytotoxicity of Sauchinone on BMDM was detected by CCK-8 kit. (B-G) BMDM were stimulated with LPS plus IFN-γ to induce M1 polarization in the presence or absence of different doses of Sauchinone. The transcription levels of Tnf-α , Il-6 , Il −12 , Inos , Il-1β , Cox-2 and Nlrp3 (B) were detected by RT-qPCR. TNF-α, IL-6, IL-12 and NO levels in culture supernatants (C) were measured by ELISA or Griess assay. The protein levels of p-STAT-1, STAT-1, and INOS in cell lysates (D) were detected by western blot. The percentages of INOS + , MHCⅡ + and CD86 + BMDM (E) were measured by FACS. (F-H) BMDM were stimulated with IL-4 plus IL-13 to induce M2 polarization in the presence or absence of different doses of Sauchinone. The transcription levels of Arg-1, Fizz1 , Ym-1 , Cd206 , Cd301 , and Dectin-1 (F) were detected by RT-qPCR. The protein levels of p-STAT-6 and STAT-6 in cell lysates (G) were detected by western blot. The percentages of CD301 + BMDM (H) were measured by FACS. Data were shown as Mean ± SEM from triplicate measurements. *P < 0.05, **P < 0.01 compared as indicated.

Journal: Journal of Advanced Research

Article Title: A novel TGR5 agonist Sauchinone ameliorates IMQ induced murine psoriasis by regulating macrophage polarization

doi: 10.1016/j.jare.2025.04.034

Figure Lengend Snippet: Sauchinone inhibits M1 macrophage polarization and promotes M2 macrophage polarization in BMDM. (A) The cytotoxicity of Sauchinone on BMDM was detected by CCK-8 kit. (B-G) BMDM were stimulated with LPS plus IFN-γ to induce M1 polarization in the presence or absence of different doses of Sauchinone. The transcription levels of Tnf-α , Il-6 , Il −12 , Inos , Il-1β , Cox-2 and Nlrp3 (B) were detected by RT-qPCR. TNF-α, IL-6, IL-12 and NO levels in culture supernatants (C) were measured by ELISA or Griess assay. The protein levels of p-STAT-1, STAT-1, and INOS in cell lysates (D) were detected by western blot. The percentages of INOS + , MHCⅡ + and CD86 + BMDM (E) were measured by FACS. (F-H) BMDM were stimulated with IL-4 plus IL-13 to induce M2 polarization in the presence or absence of different doses of Sauchinone. The transcription levels of Arg-1, Fizz1 , Ym-1 , Cd206 , Cd301 , and Dectin-1 (F) were detected by RT-qPCR. The protein levels of p-STAT-6 and STAT-6 in cell lysates (G) were detected by western blot. The percentages of CD301 + BMDM (H) were measured by FACS. Data were shown as Mean ± SEM from triplicate measurements. *P < 0.05, **P < 0.01 compared as indicated.

Article Snippet: Anti-mouse STAT-1 (9712L), anti-mouse p-STAT-1 (7649L), anti-mouse Inducible nitric oxide synthase (INOS) (13120S), Anti-mouse STAT-6 (9362S), anti-mouse p-STAT-6 (56554S), anti-mouse PKA (5842S), anti-mouse p-CREB (9197S), anti-mouse CREB (9198L),anti-mouse p-IκB (9246L), anti-mouse IκB (4812S), anti-mouse p-p65 (3033L), anti-mouse p65 (4764S), anti-mouse ASC (67824S) and anti-mouse NLRP3 (15101S) were obtained from Cell Signaling Technology (Danvers, MA, USA).

Techniques: CCK-8 Assay, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Griess Assay, Western Blot

Sauchinone inhibits macrophage M1 polarization in TGR5 dependent manner. (A-B) BMDM from WT mice were stimulated with LPS plus IFN-γ to induce M1 polarization in the presence or absence of Sauchinone (30 μM) or SBI-115 (5 μM). The transcription levels of Tnf-α , Il-6 , Inos and Il-1β (A) were detected by RT-qPCR. The protein levels of p-STAT-1, STAT-1 and INOS in cell lysates (B) were detected by western blot. (C-E) BMDM from WT mice were stimulated with LPS plus IFN-γ to induce M1 polarization in the presence or absence of Sauchinone (30 μM) or TGR5 si-RNA. The transcription levels of Tnf-α , Il-6 , Inos and Il-1β (C) were detected by RT-qPCR. The protein levels of p-STAT-1, STAT-1 and INOS in cell lysates (D) were detected by western blot. IL-6 and NO levels in culture supernatants (E) were measured by ELISA and Griess assay. (F-H) BMDM from Tgr5 -/- mice were stimulated with LPS plus IFN-γ to induce M1 polarization in the presence or absence of Sauchinone (30 μM). IL-6 and NO levels in culture supernatants (F) were measured by ELISA or Griess assay. The transcription levels of Tnf-α , Il-6 , Inos and Il-1β (G) were detected by RT-qPCR. The protein levels of p-STAT-1, STAT-1 and INOS in cell lysates (H) were detected by western blot. Data were shown as Mean ± SEM from triplicate measurements. *P < 0.05, **P < 0.01 compared as indicated.

Journal: Journal of Advanced Research

Article Title: A novel TGR5 agonist Sauchinone ameliorates IMQ induced murine psoriasis by regulating macrophage polarization

doi: 10.1016/j.jare.2025.04.034

Figure Lengend Snippet: Sauchinone inhibits macrophage M1 polarization in TGR5 dependent manner. (A-B) BMDM from WT mice were stimulated with LPS plus IFN-γ to induce M1 polarization in the presence or absence of Sauchinone (30 μM) or SBI-115 (5 μM). The transcription levels of Tnf-α , Il-6 , Inos and Il-1β (A) were detected by RT-qPCR. The protein levels of p-STAT-1, STAT-1 and INOS in cell lysates (B) were detected by western blot. (C-E) BMDM from WT mice were stimulated with LPS plus IFN-γ to induce M1 polarization in the presence or absence of Sauchinone (30 μM) or TGR5 si-RNA. The transcription levels of Tnf-α , Il-6 , Inos and Il-1β (C) were detected by RT-qPCR. The protein levels of p-STAT-1, STAT-1 and INOS in cell lysates (D) were detected by western blot. IL-6 and NO levels in culture supernatants (E) were measured by ELISA and Griess assay. (F-H) BMDM from Tgr5 -/- mice were stimulated with LPS plus IFN-γ to induce M1 polarization in the presence or absence of Sauchinone (30 μM). IL-6 and NO levels in culture supernatants (F) were measured by ELISA or Griess assay. The transcription levels of Tnf-α , Il-6 , Inos and Il-1β (G) were detected by RT-qPCR. The protein levels of p-STAT-1, STAT-1 and INOS in cell lysates (H) were detected by western blot. Data were shown as Mean ± SEM from triplicate measurements. *P < 0.05, **P < 0.01 compared as indicated.

Article Snippet: Anti-mouse STAT-1 (9712L), anti-mouse p-STAT-1 (7649L), anti-mouse Inducible nitric oxide synthase (INOS) (13120S), Anti-mouse STAT-6 (9362S), anti-mouse p-STAT-6 (56554S), anti-mouse PKA (5842S), anti-mouse p-CREB (9197S), anti-mouse CREB (9198L),anti-mouse p-IκB (9246L), anti-mouse IκB (4812S), anti-mouse p-p65 (3033L), anti-mouse p65 (4764S), anti-mouse ASC (67824S) and anti-mouse NLRP3 (15101S) were obtained from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, Griess Assay

Sauchinone inhibits macrophage M1 polarization through TGR5/PKA pathway. (A-C) BMDM were stimulated with LPS plus IFN-γ to induce M1 polarization in the presence or absence of different doses of Sauchinone (30 μM) or PKA inhibitor H89 (10 μM). IL-6 and NO levels in culture supernatants (A) were measured by ELISA or Griess assay. The transcription levels of Tnf-α , Il-6 , Inos and Il-1β (B) were detected by RT-qPCR. The protein levels of p-STAT-1, STAT-1 and INOS in cell lysates (C) were detected by western blot. Data were shown as Mean ± SEM from triplicate measurements. *P < 0.05, **P < 0.01 compared as indicated.

Journal: Journal of Advanced Research

Article Title: A novel TGR5 agonist Sauchinone ameliorates IMQ induced murine psoriasis by regulating macrophage polarization

doi: 10.1016/j.jare.2025.04.034

Figure Lengend Snippet: Sauchinone inhibits macrophage M1 polarization through TGR5/PKA pathway. (A-C) BMDM were stimulated with LPS plus IFN-γ to induce M1 polarization in the presence or absence of different doses of Sauchinone (30 μM) or PKA inhibitor H89 (10 μM). IL-6 and NO levels in culture supernatants (A) were measured by ELISA or Griess assay. The transcription levels of Tnf-α , Il-6 , Inos and Il-1β (B) were detected by RT-qPCR. The protein levels of p-STAT-1, STAT-1 and INOS in cell lysates (C) were detected by western blot. Data were shown as Mean ± SEM from triplicate measurements. *P < 0.05, **P < 0.01 compared as indicated.

Article Snippet: Anti-mouse STAT-1 (9712L), anti-mouse p-STAT-1 (7649L), anti-mouse Inducible nitric oxide synthase (INOS) (13120S), Anti-mouse STAT-6 (9362S), anti-mouse p-STAT-6 (56554S), anti-mouse PKA (5842S), anti-mouse p-CREB (9197S), anti-mouse CREB (9198L),anti-mouse p-IκB (9246L), anti-mouse IκB (4812S), anti-mouse p-p65 (3033L), anti-mouse p65 (4764S), anti-mouse ASC (67824S) and anti-mouse NLRP3 (15101S) were obtained from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Griess Assay, Quantitative RT-PCR, Western Blot

Sauchinone normalizes TGR5/PKA/CREB pathway and regulates M1/M2 macrophages markers in skin of IMQ-induced psoriasis mice. (A) The protein levels of TGR5, PKA. p-CREB, CREB in skin tissues were detected by western blot. (B) Cytokines including IL-6, IL-12 TNF-α, IFN-γ, IL-17 and IL-10 levels in serum were measured by ELISA (n = 6). (C) Cytokines including IL-6, IL-12, TNF-α, IFN-γ, IL-1β and IL-10 levels in skin homogenates were measured by ELISA (n = 6). (D) The mRNA levels of M2 markers including Ym-1 , Cd-301 , Dectin-1 , Arg-1 , Irf-4 and Ppar-γ in skin tissues were measured by qPCR (n = 6). (E) The protein levels of M1 markers including p-STAT-1, STAT1, p-p65, p65, NLRP3, p-IκB, IκB, Pro Caspase-1 and ASC in skin tissues were detected by western blot. Data were shown as Mean ± SEM from triplicate measurements. *P < 0.05, **P < 0.01 compared as indicated.

Journal: Journal of Advanced Research

Article Title: A novel TGR5 agonist Sauchinone ameliorates IMQ induced murine psoriasis by regulating macrophage polarization

doi: 10.1016/j.jare.2025.04.034

Figure Lengend Snippet: Sauchinone normalizes TGR5/PKA/CREB pathway and regulates M1/M2 macrophages markers in skin of IMQ-induced psoriasis mice. (A) The protein levels of TGR5, PKA. p-CREB, CREB in skin tissues were detected by western blot. (B) Cytokines including IL-6, IL-12 TNF-α, IFN-γ, IL-17 and IL-10 levels in serum were measured by ELISA (n = 6). (C) Cytokines including IL-6, IL-12, TNF-α, IFN-γ, IL-1β and IL-10 levels in skin homogenates were measured by ELISA (n = 6). (D) The mRNA levels of M2 markers including Ym-1 , Cd-301 , Dectin-1 , Arg-1 , Irf-4 and Ppar-γ in skin tissues were measured by qPCR (n = 6). (E) The protein levels of M1 markers including p-STAT-1, STAT1, p-p65, p65, NLRP3, p-IκB, IκB, Pro Caspase-1 and ASC in skin tissues were detected by western blot. Data were shown as Mean ± SEM from triplicate measurements. *P < 0.05, **P < 0.01 compared as indicated.

Article Snippet: Anti-mouse STAT-1 (9712L), anti-mouse p-STAT-1 (7649L), anti-mouse Inducible nitric oxide synthase (INOS) (13120S), Anti-mouse STAT-6 (9362S), anti-mouse p-STAT-6 (56554S), anti-mouse PKA (5842S), anti-mouse p-CREB (9197S), anti-mouse CREB (9198L),anti-mouse p-IκB (9246L), anti-mouse IκB (4812S), anti-mouse p-p65 (3033L), anti-mouse p65 (4764S), anti-mouse ASC (67824S) and anti-mouse NLRP3 (15101S) were obtained from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay