Journal: CNS neuroscience & therapeutics

Article Title: Oral 7,8-Dihydroxyflavone Protects Retinal Ganglion Cells by Modulating the Gut-Retina Axis and Inhibiting Ferroptosis via the Indoleacrylic Acid-AhR-ALDH1A3-FSP1 Pathway.

doi: 10.1111/cns.70442

Figure Lengend Snippet: FIGURE 5 | IDA inhibits ferroptosis in retinal neurons via the AhR-ALDH1A3-FSP1 pathway. (a) Representative western blot images of f AhR-ALDH1A3-FSP1 pathway proteins. (b–d) Quantification of nAhR, ALDH1A3, and FSP1 levels relative to β-actin. IDA increased nAhR and ALDH1A3 levels, effects partially reversed by the AhR inhibitor (iAhR) but unaffected by ALDH1A3 or FSP1 inhibitors. FSP1 levels remained un- changed. (e–g) Quantification of oxidative stress markers. IDA reduced 4-HNE and MDA levels and increased GSH levels, effects negated by iAhR, iALDH1A3, or iFSP1. Data are mean ± SEM (n = 3). One-way ANOVA with Tukey's post hoc test for (d); unpaired t-test for (b, c, e–g). p values were adjusted for multiple testing by the Benjamini Hochberg method. **p < 0.01, ***p < 0.001 vs. control; ##p < 0.01, ###p < 0.001 vs. IDA group; ns (above bars), p > 0.05 vs. ONC + DHF; ns (above horizontal lines), p > 0.05 among groups below the line.

Article Snippet: IDA's impact on ferroptosis proteins and AhR, ALDH1A3, and FSP1 inhibitors was examined in vitro and in vivo using rabbit anti- AhR (1:1000, #SAB4500725, Sigma), rabbit anti- ALDH1A3 (1:1000, #ABN427, Sigma), rabbit anti- FSP1 (1:1000, #A06541- 2, Boster), rabbit anti- GPX4 (1:1000, #SAB5700944, Sigma), rabbit anti- ACSL4 (1:2000, #SAB2100035, Sigma), rabbit anti- SLC7A11 (1:1000, #SAB5700735, Sigma), rabbit anti- GCH1 (1:1000, #PA5- 103865, Invitrogen), rabbit anti- FTH1 (1:500, #ZRB2695, Sigma), rabbit anti- DHODH (1:1000, #SAB2100574, Sigma), rabbit anti- 4- HNE (1:1000, #MA5- 27570, Invitrogen), and rabbit anti- β- actin (1:2000, #AF5003, Beyotime).

Techniques: Western Blot, Control

Journal: CNS neuroscience & therapeutics

Article Title: Oral 7,8-Dihydroxyflavone Protects Retinal Ganglion Cells by Modulating the Gut-Retina Axis and Inhibiting Ferroptosis via the Indoleacrylic Acid-AhR-ALDH1A3-FSP1 Pathway.

doi: 10.1111/cns.70442

Figure Lengend Snippet: FIGURE 6 | Neuroprotective effects of 7,8-DHF and the gut microbiota-IDA-AhR-ALDH1A3-FSP1 pathway on RGC survival and retinal function following ONC injury. (a) Retinal flat-mount images of RBPMS-labeled RGCs. Scale bar: 100 μm. (b) Representative PhNR traces showing retinal function. (c) Quantification of RGC survival (%). RGC density was reduced in the ONC group compared to the Sham group, partially preserved by 7,8-DHF, attenuated by antibiotics, and restored by IDA. AhR, ALDH1A3, and FSP1 inhibitors partially reduced RGC survival. (d) Quantification of PhNR amplitudes (μV). Retinal function decreased in the ONC group, was partially restored by 7,8-DHF, diminished by antibiotics, and recov- ered by IDA. AhR, ALDH1A3, and FSP1 inhibitors partially reduced the recovery. Data are mean ± SD (n = 6). Unpaired t-test was used except for Sham vs. ONC in (c), where Welch's t-test was applied; +++p < 0.001 vs. Sham; *p < 0.05, ***p < 0.001 vs. ONC; #p < 0.05, ##p < 0.01, ###p < 0.001 vs. ONC + DHF; &&p < 0.01, &&&p < 0.001 vs. ONC + DHF + Abx.

Article Snippet: IDA's impact on ferroptosis proteins and AhR, ALDH1A3, and FSP1 inhibitors was examined in vitro and in vivo using rabbit anti- AhR (1:1000, #SAB4500725, Sigma), rabbit anti- ALDH1A3 (1:1000, #ABN427, Sigma), rabbit anti- FSP1 (1:1000, #A06541- 2, Boster), rabbit anti- GPX4 (1:1000, #SAB5700944, Sigma), rabbit anti- ACSL4 (1:2000, #SAB2100035, Sigma), rabbit anti- SLC7A11 (1:1000, #SAB5700735, Sigma), rabbit anti- GCH1 (1:1000, #PA5- 103865, Invitrogen), rabbit anti- FTH1 (1:500, #ZRB2695, Sigma), rabbit anti- DHODH (1:1000, #SAB2100574, Sigma), rabbit anti- 4- HNE (1:1000, #MA5- 27570, Invitrogen), and rabbit anti- β- actin (1:2000, #AF5003, Beyotime).

Techniques: Labeling

Journal: CNS neuroscience & therapeutics

Article Title: Oral 7,8-Dihydroxyflavone Protects Retinal Ganglion Cells by Modulating the Gut-Retina Axis and Inhibiting Ferroptosis via the Indoleacrylic Acid-AhR-ALDH1A3-FSP1 Pathway.

doi: 10.1111/cns.70442

Figure Lengend Snippet: FIGURE 7 | Effects of 7,8-DHF, gut microbiota disruption, and IDA on AhR-ALDH1A3-FSP1 pathway proteins and oxidative stress in retinal tissues following ONC injury. (a) Representative western blot images for AhR-ALDH1A3-FSP1 pathway proteins. (b–d) Quantification of nAhR, ALDH1A3, and FSP1 levels relative to β-actin. 7,8-DHF increased nAhR and ALDH1A3 levels, which were reduced by antibiotics and restored by IDA. AhR inhibitors decreased nAhR and ALDH1A3, while ALDH1A3 or FSP1 inhibition had no effect on nAhR. FSP1 levels remained unchanged across groups. (e–g) Oxidative stress markers: 7,8-DHF reduced 4-HNE and MDA while increasing GSH levels, effects reversed by antibiotics and restored by IDA. Inhibitors of AhR, ALDH1A3, and FSP1 partially reversed these effects, implicating the AhR-ALDH1A3-FSP1 pathway. Data are mean ± SEM (n = 3). p values were adjusted for multiple testing by the Benjamini Hochberg method. *p < 0.05, **p < 0.01 vs. ONC; #p < 0.05 vs. ONC + DHF; &p < 0.05, &&p < 0.01 vs. ONC + DHF + Abx; ns (above bars), p > 0.05 vs. ONC + DHF; ns (above horizontal lines), p > 0.05 among groups below the line.

Article Snippet: IDA's impact on ferroptosis proteins and AhR, ALDH1A3, and FSP1 inhibitors was examined in vitro and in vivo using rabbit anti- AhR (1:1000, #SAB4500725, Sigma), rabbit anti- ALDH1A3 (1:1000, #ABN427, Sigma), rabbit anti- FSP1 (1:1000, #A06541- 2, Boster), rabbit anti- GPX4 (1:1000, #SAB5700944, Sigma), rabbit anti- ACSL4 (1:2000, #SAB2100035, Sigma), rabbit anti- SLC7A11 (1:1000, #SAB5700735, Sigma), rabbit anti- GCH1 (1:1000, #PA5- 103865, Invitrogen), rabbit anti- FTH1 (1:500, #ZRB2695, Sigma), rabbit anti- DHODH (1:1000, #SAB2100574, Sigma), rabbit anti- 4- HNE (1:1000, #MA5- 27570, Invitrogen), and rabbit anti- β- actin (1:2000, #AF5003, Beyotime).

Techniques: Disruption, Western Blot, Inhibition