Nrf2 activator peptide protects the brain from cerebral vascular dysfunction in alcohol ingestion
Bibhuti Ballav Saikia, Saleena Alikunju, Yemin A. Poovanthodi, Zayan Kassim, P.M. Abdul Muneer
Bibhuti Ballav Saikia, Saleena Alikunju, Yemin A. Poovanthodi, Zayan Kassim, P.M. Abdul Muneer
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Research Article Neuroscience Vascular biology

Nrf2 activator peptide protects the brain from cerebral vascular dysfunction in alcohol ingestion

Abstract

Oxidative signaling is a central mechanism in alcohol-induced injury and has strong implications for blood-brain barrier (BBB) dysregulation and neuroinflammation. Here, by targeting oxidative signaling, we hypothesized an innovative approach to develop a clinically relevant therapeutic strategy for alleviating alcohol-mediated neurovascular damage. To accomplish this, we enhanced the endogenous activity of nuclear factor E2–related factor 2 (Nrf2) by treatment with a Nrf2 activator III TAT peptide (Nrf2 peptide [NP]) and investigated the neuroprotective role of Nrf2 in promoting antioxidant defense properties and reducing BBB damage and transmigration of leukocytes to the brain following alcohol ingestion. We administered the NP subcutaneously to alcohol-ingested mice and evaluated its therapeutic potential in alleviating alcohol-associated neurovascular impairments. We compared the results with those seen in animals treated with control peptide (random sequence with TAT). The studies showed that the NP treatment preserved the oxidant-antioxidant balance, downregulated ICAM-1 and its receptors, and mitigated BBB damage and leukocyte infiltration into the brain. We validated the effect of the NP in Nrf2-knockout (Nrf2−/−) mice. Thus, this study demonstrates that NP exerts neurovascular protective effects by regulating the oxidant-antioxidant balance, reducing oxidative stress–induced BBB disruption, and limiting transmigration of immune cells to the brain in a mouse model of alcohol ingestion.

Authors

Bibhuti Ballav Saikia, Saleena Alikunju, Yemin A. Poovanthodi, Zayan Kassim, P.M. Abdul Muneer

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Figure 7

Nrf2 peptide protects the BBB in alcohol ingestion.

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Nrf2 peptide protects the BBB in alcohol ingestion. (A) Immunofluorescen...
(A) Immunofluorescence staining of claudin-5 (red) colocalized with vWF (green) and merged with DAPI (blue) in the cross section of intact brain microvessels of mice fed with CD or ED with CP or NP treatments. Scale bar: 25 μm. (B) Quantification of claudin-5 staining analyzed using ImageJ software (n = 8 per group). (C) Immunofluorescence staining of occludin colocalized with vWF (green) and merged with DAPI (blue) in the intact brain microvessels of animals fed with CD or ED with CP or NP treatments. Scale bar: 10 μm. (E–H) Western blot analysis of claudin-5, occludin, JAM-A, and β-actin from the mouse cortex tissue lysate of CD- and ED-fed animals in WT and Nrf2–/– groups with CP or NP treatments. Bar graphs with dot plot show the ratio densitometry of claudin-5 (F), occludin (G), and JAM-A (H) to β-actin (n = 8 per group). All values are expressed as mean ± SD. Statistical analysis was performed by 2-way ANOVA (B and D) and 3-way ANOVA (F and G) followed by Bonferroni’s post hoc test. Statistically significant, *P < 0.05, **P < 0.01, ***P < 0.001 vs. their representative control groups (CD) in B and D. @P < 0.05, @@P < 0.01, @@@P < 0.001 vs. representative groups in WT animals. P < 0.05 was considered statistically significant. Exact P values are shown between the compared groups in F–H.