The protection of sulforaphane on subarachnoid hemorrhage-induced intestinal mucosa injury in rats

萝卜硫素对大鼠蛛网膜下腔出血引起的肠黏膜损伤的保护作用

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Abstract

INTRODUCTION: Sulforaphane (SFN) is recognized for its anti-inflammatory properties; however, the underlying molecular mechanisms remain unclear. In this study, we explored the effect of SFN on subarachnoid hemorrhage (SAH) and the potential mechanisms. METHODS: Sprague-Dawley (SD) rats were divided into three groups (n = 12): Sham + vehicle group (Sham + V), SAH + vehicle group (SAH + V), and SAH + SFN group (SAH + S). SFN (50 mg/kg) dissolved in 250-280 μL corn oil was intraperitoneally injected, and the same volume of corn oil was served as the control. The appetite score, gut wet/dry weight ratio, and histological changes in ileum tissues were examined to determine intestinal mucosal injury. Quantitative real-time PCR (qRT-PCR) and Western blot were performed to examine the expression of genes. LC3 immunofluorescence and Hoechst 33258 staining were used to assess cell autophagy and apoptosis. RESULTS: Compared to the SAH + V group, the SAH + S group demonstrated a significantly increased appetite score (1.55 ± 0.23 vs. 1.90 ± 0.35); decreased gut wet/dry weight ratio (4.02 ± 0.21 vs. 3.18 ± 0.21) and inflammatory score (2.89 ± 0.33 vs. 1.89 ± 0.60); elevated mRNA expression of Nrf-2 (1.12 ± 0.14 vs. 1.89 ± 0.12), HO-1 (0.46 ± 0.02 vs. 1.02 ± 0.10), and NQO-1 (1.35 ± 0.09 vs. 1.97 ± 0.18); and elevated protein levels of Nrf-2 (0.92 ± 0.18 vs. 1.43 ± 0.23), Keap1 (0.31 ± 0.03 vs. 0.44 ± 0.02), HO-1 (0.65 ± 0.02 vs. 0.88 ± 0.02), NQO-1 (0.58 ± 0.02 vs. 0.78 ± 0.02), LC3-II/I (0.20 ± 0.004 vs. 0.28 ± 0.01), ATG4D (0.45 ± 0.01 vs. 0.72 ± 0.04), and P62 (0.85 ± 0.01 vs. 0.99 ± 0.03). The in vitro experiments further revealed that 3-methyladenine (3-MA) significantly reversed the decreased apoptosis of IEC-6 cells induced by 20 μmol/L SFN (20.60 ± 1.28 vs. 11.50 ± 0.58). CONCLUSION: SFN exhibited the protective effect on intestinal mucosa injury after SAH via activating autophagy, which may provide an innovative approach to alleviate the intestinal mucosa injury caused by SAH.

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