A Multi-Mineral Intervention to Modulate Colonic Mucosal Protein Profile: Results from a 90-Day Trial in Human Subjects

Abstract

The overall goal of this study was to determine whether Aquamin®, a calcium-, magnesium-, trace element-rich, red algae-derived natural product, would alter the expression of proteins involved in growth-regulation and differentiation in colon. Thirty healthy human subjects (at risk for colorectal cancer) were enrolled in a three-arm, 90-day interventional trial. Aquamin® was compared to calcium alone and placebo. Before and after the interventional period, colonic biopsies were obtained. Biopsies were evaluated by immunohistology for expression of Ki67 (proliferation marker) and for CK20 and p21 (differentiation markers). Tandem mass tag-mass spectrometry-based detection was used to assess levels of multiple proteins. As compared to placebo or calcium, Aquamin® reduced the level of Ki67 expression and slightly increased CK20 expression. Increased p21 expression was observed with both calcium and Aquamin®. In proteomic screen, Aquamin® treatment resulted in many more proteins being upregulated (including pro-apoptotic, cytokeratins, cell–cell adhesion molecules, and components of the basement membrane) or downregulated (proliferation and nucleic acid metabolism) than placebo. Calcium alone also altered the expression of many of the same proteins but not to the same extent as Aquamin®. We conclude that daily Aquamin® ingestion alters protein expression profile in the colon that could be beneficial to colonic health.

Cell-matrix adhesion contributes to permeability control in human colon organoids

“…Equally important, when Aquamin ® -treated subjects were compared to subjects receiving placebo for the same period, we saw up-regulation of laminin chains along with increased levels of other basement membrane components and hemidesmosome moieties in colonic biopsies (46). Subjects receiving calcium alone (i.e., the most abundant mineral in Aquamin ® ) also demonstrated increases in several of the same molecules, but the degree of up-regulation with calcium alone was lower than that seen with Aquamin ® (46).…”

“…Subjects receiving calcium alone (i.e., the most abundant mineral in Aquamin ® ) also demonstrated increases in several of the same molecules, but the degree of up-regulation with calcium alone was lower than that seen with Aquamin ® (46).…”

“…To summarize the results of this pilot study, there were no tolerability issues with daily Aquamin ® ingestion over a 90-day period and no safety concerns (45). Equally important, when Aquamin ® -treated subjects were compared to subjects receiving placebo for the same period, we saw up-regulation of laminin chains along with increased levels of other basement membrane components and hemidesmosome moieties in colonic biopsies (46).…”

Liver Protein Expression in Nash Mice on a High-Fat Diet

“…We have recently completed a 90-day pilot phase trial in which 30 healthy human subjects were randomized to receive Aquamin ® formulated to provide 800 mg of calcium per day, calcium carbonate at the same level or placebo (37,77). To summarize, no safety or tolerability issues were seen with Aquamin ® .…”

“…For protein mass spectrometry analysis, we employed liver tissue from five mice from each group and assessed each separately. A cryopreserved tissue piece from the left lobe (of each mouse liver) was weighed and homogenized in Radioimmuno-Precipitation Assay (RIPA) -lysis and extraction buffer (Pierce, # 89901; ThermoFisher Scientific) for protein isolation, as described in our previous publications (37)(38)(39). Fifty micrograms of sample protein from each liver were digested separately with trypsin and individual samples labeled with one of 11 isobaric mass tags following the manufacturer's protocol.…”