In vitro cytotoxic and genotoxic effects of Cissus verticillata and Sphagneticola trilobata used for treatment of Diabetes Mellitus in Brazilian folk medicine

Cissus verticillata and Sphagneticola trilobata have been used in Brazilian folk medicine for Diabetes Mellitus treatment, although their pharmacological and toxicological profile has not been clearly established. Thus, the aim of this study was to evaluate the preclinical toxicity of the aqueous extracts of C. verticillata and S. trilobata. The main groups of secondary metabolites were investigated, and the species differed by the presence of coumarins in C. verticillata and by tannins in S. trilobata extracts. The highest contents of phenolic compounds and flavonoids were quantified in C. verticillata infusion with 2.594 ± 0.04 mg equivalents of gallic acid g-1 of extract and 1.301 ± 0.015 mg equivalents of catechin g-1 of extract, respectively. While the extract of S. trilobata showed minimum values of these compounds, with 0.002 ± 0.001 mg equivalents of gallic acid g-1 extract and 0.005 ± 0.0004 mg equivalents of catechin g-1 of extract, respectively. These differences implied the results of in vitro antioxidant activity evaluated using ferric reducing antioxidant power (FRAP), in which the sample of C. verticillata at 5 mg mL-1 showed a value of 122 µM ferrous sulfate equivalents (FSE), while S. trilobata showed 0.93 µM FSE at the same concentration. With respect to cytotoxic assay with murine fibroblast cell line (3T3) only S. trilobata exhibited cytotoxic effects measured by MTT and Sulforhodamine B assays, evidenced by the cell viability value of approximately 16%, in both tests after 24 and 72 hours of exposure of the cells to 5 mg mL-1 of the extract. Comparatively, at 5 mg mL-1 the C. verticillata extract showed cell viability of 142% and 95%, respectively, after 24 hours of cell exposure. On the other hand, both species showed genotoxic profiles evidenced by chromosomal aberrations by Allium cepa bioassay, observed by the higher percentage values of chromosome bridges, chromosome loss, and disturbed anaphase for all concentrations of both extracts than those of the negative control. The results support the characterization of the toxicological profile for both species and create an alert regarding the use of S. trilobata, which should be avoided.

Influence of rare earth elements on metabolism and related enzyme activity and isozyme expression in Tetrastigma hemsleyanum cell suspension cultures.

The effects of rare earth elements (REEs) not only on cell growth and flavonoid accumulation of Tetrastigma hemsleyanum suspension cells but also on the isoenzyme patterns and activities of related enzymes were studied in this paper. There were no significant differences in enhancement of flavonoid accumulation in T. hemsleyanum suspension cells among La(3+), Ce(3+), and Nd(3+). Whereas their inductive effects on cell proliferation varied greatly. The most significant effects were achieved with 100 µM Ce(3+)and Nd(3+). Under treatment over a 25-day culture period, the maximal biomass levels reached 1.92- and 1.74-fold and the total flavonoid contents are 1.45- and 1.49-fold, than that of control, respectively. Catalase, phenylalanine ammonia-lyase (PAL), and peroxidase (POD) activity was activated significantly when the REE concentration range from 0 to 300 µM, whereas no significant changes were found in superoxide dismutase activity. Differences of esterase isozymes under REE treatment only laid in expression level, and there were no specific bands. The expression level of some POD isozymes strengthened with increasing concentration of REEs within the range of 50-200 µM. When REE concentration was higher than 300 µM, the expression of some POD isozymes was inhibited; meanwhile, some other new POD isozymes were induced. Our results also showed REEs did not directly influence PAL activity. So, we speculated that 50-200 µM REEs could activate some of antioxidant enzymes, adjust some isozymes expression, trigger the defense responses of T. hemsleyanum suspension cells, and stimulate flavonoid accumulation by inducing PAL activity.

[Macroscopic and microscopic identification of Zhuang medicine Tetrastigma planicaule].

OBJECTIVE: To study the macroscopic and microscopic characteristics of Zhuang medicine Tetrastigma planicaule. METHODS: Macroscopic and microscopic identification were studied. RESULTS: The microscopic characteristics were significant. The root had more than 10 layers of cork cells arranging in line. The starch grain and calcium oxalate cluster crystals distributed in the phloem parenchymas, and mucilage cells scattered in the cortex of the stem. There were 8 vascular bundles in the vein of leaves. CONCLUSION: This study provides a scientific basis for the identification and utilizing of Tetrastigma planicaule.

Structural and immunocytochemical characterization of the adhesive tendril of Virginia creeper (Parthenocissus quinquefolia [L.] Planch.).

The tendrils of Virginia creeper (Parthenocissus quinquefolia) do not coil around their supports. Rather, they adhere to supporting objects by flattening against the support surface and secreting an adhesive compound which firmly glues the tendril to the support. In this study, microscopic and immunocytochemical techniques were utilized to determine the nature of this adhesive. Following touch stimulation, epidermal cells of the tendril elongate toward the support substrate, becoming papillate in morphology. Following contact with the support surface, an adhesive is produced at the base of the papillate cells. The adhesive appears as a highly heterogeneous, raftlike structure and consists of pectinaceous, rhamnogalacturonan (RG) I-reactive components surrounding a callosic core. In addition, more mobile components, composed of arabinogalactans and mucilaginous pectins, intercalate both the support and the tendril, penetrating the tendril to the proximal ends of the papillate cells. Following adherence to the support, the anticlinal walls of the papillate cells are devoid of RG I side-chain reactivity, indicating that extensive debranching of RG I molecules has taken place. Furthermore, a large amount of RG I backbone reactivity was observed in the contact area. These results may indicate that the debranched RG I molecules diffuse into and permeate the contact region, forming an integral part of the adhesive compound. These results indicate that Virginia creeper adheres to objects by a composite adhesive structure consisting of debranched RG I, callose, and other, less-well characterized mucilaginous pectins and that this structure subsequently becomes lignified and very weather-resistant upon the ultimate senescence of the tendril.