Phytochemical Combination Is More Effective than Individual Components in Reducing Stress Signaling in Rat Hippocampal Neurons and Microglia In Vitro

Int J Mol Sci. 2022 Oct 21;23(20):12651. doi: 10.3390/ijms232012651.

Abstract

Age-related decrements in the central nervous system (CNS) are thought to result from: (1) increased susceptibility to and accumulating effects of free radicals and inflammation; and (2) dysregulation in Ca2+ homeostasis, which affects numerous signaling pathways. Certain bioactive phytochemicals exhibit potent anti-inflammatory activities which may mitigate these age-related CNS decrements. This study investigated the individual and combination effects of green tea catechin (epigallocatechin gallate, EGCG), curcumin from turmeric, and broccoli sprouts which contain the isothiocyanate sulforaphane on inflammation and dysregulation in Ca2+ homeostasis to determine if the individual compounds were working synergistically and/or through independent mechanisms. Rat hippocampal neurons or highly aggressive proliferating immortalized (HAPI) microglial cells were pre-treated for a week with either the individual components or all in combination before inducing Ca2+ buffering deficits with dopamine (DA, 0.1 µM for 2 h) or inflammation using lipopolysaccharide (LPS, 100 ng/mL for 18 h), respectively. The EGCG (3 µM) and combination protected against DA-induced deficits in Ca2+ buffering (both % of cells that recovered and recovery time, p < 0.05). Additionally, the EGCG and combination reduced stress-mediated inflammation in HAPI rat microglial cells by attenuating LPS-induced nitrite release, inducible nitrous oxide synthase (iNOS) expression, and tumor necrosis factor-alpha (TNF-α) release (p < 0.05), but not cyclooxygenase-2 (COX-2) expression. Overall, broccoli sprouts (2 µM) and curcumin (1 µM) were not as effective as the EGCG or combination. Further research is needed to determine if dietary intervention with a variety of foods containing compounds such as those found in green tea, turmeric, or broccoli sprouts can play a role in reducing age-related CNS inflammation, microglial activation, and downstream signaling pathways that can lead to neuronal dysfunction.

Keywords: EGCG; anti-inflammatory; broccoli sprouts; calcium signaling; curcumin; microglia; neurons; sulforaphane.

MeSH terms

  • Animals
  • Anti-Inflammatory Agents / pharmacology
  • Catechin* / therapeutic use
  • Curcumin* / therapeutic use
  • Cyclooxygenase 2 / metabolism
  • Dopamine / metabolism
  • Hippocampus / metabolism
  • Inflammation / pathology
  • Isothiocyanates / therapeutic use
  • Lipopolysaccharides / pharmacology
  • Microglia / metabolism
  • Neurons / metabolism
  • Nitrites / metabolism
  • Nitrous Oxide / adverse effects
  • Nitrous Oxide / metabolism
  • Phytochemicals / therapeutic use
  • Rats
  • Tea / metabolism
  • Tumor Necrosis Factor-alpha / metabolism

Substances

  • Catechin
  • Lipopolysaccharides
  • Tumor Necrosis Factor-alpha
  • Curcumin
  • Dopamine
  • Cyclooxygenase 2
  • Nitrites
  • Nitrous Oxide
  • Isothiocyanates
  • Tea
  • Anti-Inflammatory Agents
  • Phytochemicals

Grants and funding

Source of Support: USDA ARS Project Number 8050-51000-102-000-D.