Chlorogenic Acid and Biohanin A from Trifolium pratense L. Callus Culture Extract: Functional Activity In Vivo

Abstract

Polyphenols are potential neuroprotectors that increase lifespan and slow down aging. Red clover (Trifolium pratense L.) is a promising source of biologically active substances. Its extracts contain biochanin A and chlorogenic acid. This research used Caenorhabditis elegans to study the effect of polyphenols extracted from red clover callus cultures on SOD-3 and HSP-16.2 genes, as well as their anti-amyloid potential. 
 The chlorogenic acid and biochanin A (200, 100, 50, and 10 µM) with a purity of 95% were isolated from callus extracts of T. pratense L. The effect of polyphenols on SOD-3 and HSP-16.2 was assessed after 5 and 2 h of heat stress (35°C), respectively, using C. elegans N2 Bristol as model organism. The neuroprotective potential was measured by counting paralyzed nematodes after 18, 40, and 62 h of incubation.
 The research established a dose-dependent effect between the concentration of biologically active substances and the percentage of paralyzed nematodes after 18 h of cultivation. The lowest paralysis phenotype count occurred at a concentration of 200 μM. The activity of 200 μM biochnin A was 1.18 times as high as that of a 200 μM chlorogenic acid solution. Biochanin A solutions increased SOD-3 expression by 3.7 times, compared to the control.
 The biologically active substances exhibited relative neuroprotective activity and affected the expression of antioxidant defense gene in C. elegans.

The Biological Active Substances of Taraxacum officinale and Arctium lappa from the Siberian Federal District

“…Volumes of 2, 3, 5, 7, 10 µL of the studied extract and 3 µL of 0.1% solutions of standard samples of rutin and quercitin were simultaneously applied to the plastic. To obtain standard solutions of rutin and quercetin, stock solutions of dimethyl sulfoxide (Lenreaktiv, St Petersburg, Russia) were prepared and diluted with distilled water [ 15 ]. After TLC analysis, the spots were exposed under a lamp with ultraviolet light at 254 nm.…”

“…Antioxidants derived from plant raw materials can react with reactive oxygen species, thereby protecting cellular structures from free-radical damage and contributing to the reduction of oxidative stress [ 15 ]. Substances such as vitamins, macro- and microelements, phenolic compounds, alkaloids, carotenoids, minerals, and enzymes found in fruits, vegetables, berries, and plants fall into the category of natural antioxidants.…”