RESUMEN
Introduction: Inflammatory bowel diseases (IBDs) include Crohn's disease, and ulcerative colitis. Cannabis sativa preparations have beneficial effects for IBD patients. However, C. sativa extracts contain hundreds of compounds. Although there is much knowledge of the activity of different cannabinoids and their receptor agonists or antagonists, the cytotoxic and anti-inflammatory activity of whole C. sativa extracts has never been characterized in detail with in vitro and ex vivo colon models. Material and Methods: The anti-inflammatory activity of C. sativa extracts was studied on three lines of epithelial cells and on colon tissue. C. sativa flowers were extracted with ethanol, enzyme-linked immunosorbent assay was used to determine the level of interleukin-8 in colon cells and tissue biopsies, chemical analysis was performed using high-performance liquid chromatography, mass spectrometry and nuclear magnetic resonance and gene expression was determined by quantitative real-time PCR. Results: The anti-inflammatory activity of Cannabis extracts derives from D9-tetrahydrocannabinolic acid (THCA) present in fraction 7 (F7) of the extract. However, all fractions of C. sativa at a certain combination of concentrations have a significant increased cytotoxic activity. GPR55 receptor antagonist significantly reduces the anti-inflammatory activity of F7, whereas cannabinoid type 2 receptor antagonist significantly increases HCT116 cell proliferation. Also, cannabidiol (CBD) shows dose dependent cytotoxic activity, whereas anti-inflammatory activity was found only for the low concentration of CBD, and in a bell-shaped rather than dose-dependent manner. Activity of the extract and active fraction was verified on colon tissues taken from IBD patients, and was shown to suppress cyclooxygenase-2 (COX2) and metalloproteinase-9 (MMP9) gene expression in both cell culture and colon tissue. Conclusions: It is suggested that the anti-inflammatory activity of Cannabis extracts on colon epithelial cells derives from a fraction of the extract that contains THCA, and is mediated, at least partially, via GPR55 receptor. The cytotoxic activity of the C. sativa extract was increased by combining all fractions at a certain combination of concentrations and was partially affected by CB2 receptor antagonist that increased cell proliferation. It is suggested that in a nonpsychoactive treatment for IBD, THCA should be used rather than CBD.
RESUMEN
Strigolactones (SLs) are plant hormones and regulators of root development, including lateral root (LR) formation, root hair (RH) elongation and meristem cell number, in a MORE AXILLARY GROWTH 2 (MAX2)-dependent way. However, whether SL signaling is acting cell-autonomously or in a non-cell-autonomous way in roots is unclear. We analyzed root phenotype, hormonal responses and gene expression in multiple lines of Arabidopsis thaliana max2-1 mutants expressing MAX2 under various tissue-specific promoters and shy2 mutants. The results demonstrate for the first time that expression of MAX2 under the SCARECROW (SCR) promoter, expressed mainly in the root endodermis, is sufficient to confer SL sensitivity in the root for RH, LR and meristem cell number. Moreover, loss of function mutation of SHORT HYPOCOTYL 2 (SHY2), a key component in auxin and cytokinin regulation of meristem size, has been found to be insensitive to SLs in relation to LR formation and meristem cell number. Endodermal SL signaling, mediated by MAX2, is sufficient to confer SL sensitivity in root, and SHY2 may participate in SL signaling to regulate meristem size and LR formation. These SL signaling pathways thus may act through modulation of auxin flux in the root tip, and may indicate a root-specific, yet non-cell-autonomous regulatory mode of action.