Stress, Inflammation, and Astrodegeneration; Epicatechin And Alpha Lipoic Acid as Dietary Supplements

Date of Award

7-2019

Degree Type

Thesis

Degree Name

Master of Science in Biomedical Sciences

First Advisor

Harold Komisky, PhD

Second Advisor

Abigail Hielscher, PhD

Third Advisor

Xinyu Wang, PhD

Fourth Advisor

Richard White, PhD

Abstract

Astrocytes play a critical role in the development, function, and viability of the central nervous system. Stress induced inflammation may reduce the viability of astrocytes in vivo and in cell cultures. Although essential trace metals like manganese (Mn) are required for normal physiological processes. Increased exposure to Mn had been claimed to interfere with brain development and cognition. Researchers have argued about possible mechanisms, like mitochondrial dysfunction, inflammation, and increased free radical involvement in mediating the toxic effects of this essential trace metal. There is also a need to detect early signs of neurotoxicity from environmental and occupational exposure to Mn in order to provide a quantitative estimate of health risk.

Significant changes in glial fibrillary acidic protein (GFAP) has been argued to be a biomarker for neurotoxicity. The ability of manganese sulfate (MnS04) to increase GF AP levels in astrocytes of rats fed diets containing high levels of this metal have been reported. Supplements may be helpful in preventing this brain pathology. To assess the hypothesis that alpha lipoic acid and epicatechin increases viability of primary rat embryonic (E18) hypothalamic astrocytes, cell cultures were pretreated with epicatechin or alpha-lipoic acid prior to Mn treatment. The levels of GFAP released by the astrocytes was measured using sandwiched Enzyme-Linked lmmunosorbent Assay (ELISA). ELISA analysis indicated that alpha lipoic acid and epicatechin exposure had no significant effect on GFAP levels at 72 hrs.

In addition, a Water-Soluble Tetrazoliurn-8 (WST-8) method was utilized to determine cell viability. The WST-8 assay showed that alpha lipoic acid and epicatechin had no significant effect on dehydrogenase activity in astrocytes at 72 hrs. In summary, alpha-lipoic acid, and epicatechin as dietary supplements had no protective effect on rat E18 hypothalamic astrocytes after acute Mn over exposure.

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