ABSTRACT
A histological analysis was performed with the aim of elucidating the spontaneous regeneration process of the hairy root lines LRT 2.3 and LRT 6.4, derived from Lopezia racemosa leaf explants and genetically transformed with the Agrobacterium rhizogenes strain ATCC15834/pTDT. The analysis showed both lines regenerate via indirect somatic embryogenesis; LRT 6.4 also regenerated by direct organogenesis. The morphogenic characteristics of the regenerated plantlets from both lines showed the typical characteristics, described previously, including a higher number of axillary shoot formation, short internodes, and plagiotropic roots compared with wild-type seedlings. The regeneration process occurred without the addition of plant growth regulators and was linked to the sucrose concentration in the culture medium. Reducing the sucrose concentration from 3% to 2%, 1%, and 0.5% increased the regeneration rate in LRT 6.4; the effect was less pronounced in LRT 2.3. The cytotoxic activity of different organic extracts obtained from roots and shoots were evaluated in the cancer cell lines HeLa (cervical carcinoma), HCT-15 (colon adenocarcinoma), and OVCAR (ovary carcinoma). The hexane and dichloromethane extracts from roots of both lines showed cytotoxic activity against the HeLa cell line. Only the dichloromethane extract from the roots of PLRT 2.3 showed cytotoxic activity against the OVCAR cell line. None of the methanol extracts showed cytotoxic activity, nor the shoot extracts from any solvent.
ABSTRACT
AIM OF THE STUDY: To isolate compounds with anti-inflammatory activity from Bursera cuneata by a bioassay-guided fractionation. MATERIALS AND METHODS: Three extracts of different polarities were elaborated by maceration. These extracts were assayed for their inhibitory effects on phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) induced edema in mice. The dichloromethane extract was subjected to activity guided fractionation using successive chromatographic procedures. Additionally, the levels of histamine were determined in the ear samples obtained from the TPA assay, which were determined by high-performance liquid chromatography (HPLC). Effect of moronic Acid on RAW 264.7 stimulated with LPS was evaluated for NO and TNF secretion. RESULTS: The dichloromethane extract had the highest anti-inflammatory effect (89.1⯱â¯2.2% inhibition) over that of the hexane (53.3⯱â¯1.2%) and methanolic (77.4⯱â¯1.8%) extracts at a dose of 0.1 mg/ear. The FS-3 fraction, obtained from the dichloromethane extract, comprised triterpenes ß-sitosterol (1), α-amyrin (2), moronic acid (3), and ursolic acid (4), and all the compounds showed significant activity in comparison with that of indomethacin (41.5⯱â¯0.6%) at 0.1 mg/mouse ear. However, moronic acid displayed the highest inhibitory effect (68.1⯱â¯1.3%). Additionally, levels of histamine were determined by HPLC in the treated tissues. moronic acid was the most active (73.3⯱â¯1.1%, indomethacin 33.8⯱â¯0.8%). The bio-guided isolation resulted in the identification of moronic acid as the principal anti-inflammatory and antihistaminic compound present in B. cuneata. To confirm a general anti-inflammatory effect, moronic acid was evaluated on the activation of RAW 264.7â¯cell stimulated with LPS. At 30 and 15â¯mg/mL a significant reduction of ON was observed (36% and 28% respectively) but had no significant effect on TNFα production. CONCLUSIONS: Our study showed that the organic extracts and isolated compounds from the aerial parts of B. cuneata had topical anti-inflammatory and antihistaminic activities in vivo, but in vitro only modified the production of ON in RAW cells. The results of this study validated the use of B. cuneata in folk medicine for the treatment of inflammatory diseases.
