Phenolic Antioxidant Compounds Produced byin Vitro. Cultures of Rosemary (Rosmarinus officinalis.) and Their Anti-inflammatory Effect on Lipopolysaccharide-Activated Microglia

Abstract

Different culture types (shoot culture, callus culture, and cell suspension culture) of Rosmarinus officinalis L. were established and their ability to biosynthesize the phytochemicals carnosic acid, carnosol, and rosmarinic acid were assessed and compared with subcultures of the same stem plant over time. Furthermore, we examined the antioxidative effect of extracts from different culture types by measuring their scavenging activity on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals as well as their anti-inflammatory potential on neonatal microglial rat cells activated by means of the inflammogen lipopolysaccharide (LPS) taking nitric oxide (NO) as parameter. Our results show that during an investigation period of 17 months, the shoot culture accumulated varying amounts of carnosic acid and carnosol, which were also present in callus culture but about 20-to 80-fold lower than in the shoot culture. In suspension culture, only carnosic acid and no carnosol could be detected. The level of carnosic acid in suspension culture was threefold less than detected for the callus culture on average. The amount of rosmarinic acid produced in shoot culture and callus culture were comparable, whereas in suspension culture higher concentrations of rosmarinic acid could be measured than in shoot and callus culture. Thus, the content of carnosic acid, carnosol, and rosmarinic acid in the extracts depended on the differentiation grade of the cell culture type. The DPPH radical-scavenging activity of the extracts depended on the amount of all three phytochemicals, in particular of rosmarinic acid. The anti-inflammatory character of the extracts was mainly based on their carnosic acid content.

Schisandra rubriflora Plant Material and In Vitro Microshoot Cultures as Rich Sources of Natural Phenolic Antioxidants

“…Kuhlmann and Röhl [70] compared the phenolic content using HPLC-DAD and the antioxidant capacity with DPPH of different in vitro culture types, namely shoot culture, callus culture and cell suspension of Rosmarinus officinalis. The dominant compounds found were diterpenes (carnosic acid and carnosol) and depside (rosmarinic acid).…”

Mechanisms Involved in the Modulation of Astroglial Resistance to Oxidative Stress Induced by Activated Microglia: Antioxidative Systems, Peroxide Elimination, Radical Generation, Lipid Peroxidation

“…Flat, well attached, nonstellate-shaped microglia were considered as quiescent (Röhl and Sievers 2005), if no NO release into the culture medium could be measured by means of the Griess assay. Then, stimulation with 0.02 lg/ml lipopolysaccharides (LPS, L-8274, Sigma) in 310 ll/cm 2 growth medium was performed for 24 h. Previously, we have shown that this concentration induces an increased microglial mRNA expression of IL-6 (130-fold), IL-1b (60-fold), TNF-a (3-fold), iNOS (200-fold) and COX-2 (14-fold) (Röhl et al 2007) and a maximal LPSinduced NO production in our microglial cell cultures (Kuhlmann and Röhl 2006) but not in purified astroglial cultures (Röhl and Sievers 2005).…”

The Therapeutic Potential of Rosemary (Rosmarinus officinalis) Diterpenes for Alzheimer’s Disease

“…Moreover, both microglia and astrocytes have been shown to be highly regulated in AD brains [ 133 , 134 ]. The potent anti-inflammatory activity of rosemary diterpenes in both the microglial cells [ 126 , 135 ] and other inflammatory models therefore suggests their potential in tackling AD.…”