Phytochemical combination is more effective than individual components in reducing stress signaling in rat hippocampal neurons and microglia

Authors: Fisher, Derek; ZHENG, TONG - Tufts University; BIELINSKI, DONNA - Tufts University; KELLY, MEGAN - University Of North Texas; CAHOON, DANIELLE - Tufts University; Shukitt-Hale, Barbara

Submitted to: International Journal of Molecular Sciences
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 10/15/2022
Publication Date: 10/21/2022
Citation: Fisher, D.R., Zheng, T., Bielinski, D.F., Kelly, M.E., Cahoon, D.S., Shukitt Hale, B. 2022. Phytochemical combination is more effective than individual components in reducing stress signaling in rat hippocampal neurons and microglia. International Journal of Molecular Sciences. 23(20):12651. https://doi.org/10.3390/ijms232012651.
DOI: https://doi.org/10.3390/ijms232012651

Interpretive Summary: Age-related changes in the brain are thought to be due to oxidative stress, inflammation, and problems regulating calcium. However, certain nutrients in foods may be able to lessen some of these decreases. This study investigated the individual and combination effects of a green tea compound, the spice tumeric, and broccoli sprouts on inflammation and calcium regulation to determine if the individual compounds were working together and/or independently. Cells derived from rats were pre-treated for a week with either the individual components or all in combination before inducing calcium regulatory deficits or inflammation. The green tea compound and combination protected against stressor-induced deficits in calcium regulation and inflammation. Overall, broccoli sprouts and tumeric were not as effective as the green tea compound 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 brain inflammation and calcium dysfunction.

Technical 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, 0.1uM for 2 hours) or inflammation using lipopolysaccharide (LPS, 100 ng/ml overnight), respectively. The EGCG (3uM) 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-a) release (p < 0.05), but not cyclooxygenase-2 (COX-2) expression. Overall, broccoli sprouts (2uM) and curcumin (1uM) 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.