Evaluation of the anxiolytic activity of dry extract of Patrinia scabiosifolia under acoustic stress.

OBJECTIVE: Patrinia scabiosifolia has been used in traditional medicine in East Asia, Africa, and South America for a variety of diseases for more than 2000 years. The purpose of the article is to evaluate the anxiolytic properties of dry extract of P. scabiosifolia. MATERIALS AND METHODS: In vivo experiments were performed on outbred white male mice. The psychotropic effect of P. scabiosifolia dry extract was assessed using behavioral test systems aimed at identifying changes in the psycho-emotional state of animals under the influence of acoustic stress. In addition, the preparation toxicity was also assessed. HPLC-MS analysis was carried out to confirm the presence of active components in local raw materials. RESULTS: The article describes the possibility of using dry extract of P. scabiosifolia as an anxiolytic and sedative for psycho-emotional stress in experimental animals. Based on comprehensive research results, the effectiveness and safety of the studied herbal preparation have been proven. CONCLUSIONS: In this study, a dry extract of P. scabiosifolia has been proposed as a novel means of combating neuropsychiatric disorders. P. scabiosifolia showed efficacy comparable to the reference drug (Mebicar), reducing sleep time and increasing sleep duration. The results obtained can subsequently serve as the basis for clinical trials.

Methylated flavones of the hairy root culture Scutellaria baicalensis.

Perennial plants in northern Dauria (Zabaikalsky region) grow in low temperatures in winter and in a dry hot summer. The prairies of northern Dauria are rich in a variety of medicinal herbs, including S. baicalensis, which has roots that are in demand for traditional Chinese medicine. In addition to two monomethylated flavones (wogonin and oroxylin A), determining the pharmacological significance of the root, there is also a minority of their polymethylated congeners. Little is known about their role in the plant or their connection with the conditions of growth and cultivation of their hairy root culture (HRC). Therefore, the purpose of this study was to determine whether and to what extent the biosynthesis of the latter is retained in the hairy root culture established from wild plants of Dauria. The composition of the main methylated flavones of HRC was established using LC-MS and a previously unknown pentamethylated flavone was found in the roots. This study showed a more significant accumulation of polymethylated flavones in the root of the wild plant than in HRC.

Regulation of resveratrol production in Vitis amurensis cell cultures by calcium-dependent protein kinases.

Resveratrol is a naturally occurring plant stilbene that exhibits a wide range of valuable biological and pharmacological properties. Although the beneficial effects of trans-resveratrol to human health and plant protection against fungal pathogens are well-established, little is known about the molecular mechanisms regulating stilbene biosynthesis in plant cells. It has been recently shown that overexpression of the calcium-dependent protein kinase VaCPK20 gene considerably increased resveratrol accumulation in cell cultures of Vitis amurensis. It is possible that calcium-dependent protein kinases (CDPKs) play an important role in the regulation of resveratrol biosynthesis. In the present work, we investigated the effects of overexpression of other members of the CDPK multigene family (VaCPK9, VaCPK13, VaCPK21, and VaCPK29) on resveratrol accumulation and growth parameters of grape cell cultures. The obtained data show that overexpression of VaCPK29 increased resveratrol content 1.6-2.4-fold and fresh biomass accumulation 1.1-1.4-fold in the four independently transformed cell lines of V. amurensis compared with that in the empty vector-transformed calli. However, overexpression of the VaCPK9, VaCPK13, and VaCPK21 genes did not considerably affect resveratrol content and fresh/dry biomass accumulation in the independently transformed cell lines of V. amurensis. VaCPK29-transformed calli were capable of producing between 1.02 and 1.39 mg/l of resveratrol, while the control calli produced 0.48 to 0.79 mg/l of resveratrol. The data indicate that the VaCPK9, VaCPK13, and VaCPK21 genes are not involved in the regulation of stilbene biosynthesis in grape cells, while the VaCPK29 and VaCPK20 genes are implicated in resveratrol biosynthesis as positive regulators.

VaCPK20 gene overexpression significantly increased resveratrol content and expression of stilbene synthase genes in cell cultures of Vitis amurensis Rupr.

Resveratrol, a naturally occurring plant phenol, has been reported to exhibit a wide range of valuable biological and pharmacological properties. In the present investigation, we show that transformation of a Vitis amurensis Rupr. cell suspension with the gene VaCPK20 for a calcium-dependent protein kinase (CDPK) under the control of double CaMV 35S promoter increased resveratrol production in five independently transformed cell lines in 9-68 times compared with control cells. The VaCPK20-transformed calli were capable of producing 0.04-0.42 % dry wt. of resveratrol, while the control calli produced up to 0.008 % dry wt. of resveratrol Also, we characterized expression of stilbene synthase (STS) genes in the five VaCPK20-transgenic cell lines of V. amurensis. In all VaCPK20-transgenic cell lines, expression of VaSTS7 increased; while expression of VaSTS1 decreased. We suggest that transformation of V. amurensis calli with the VaCPK20 gene induced resveratrol accumulation via enhancement of expression of the VaSTS7 gene involved in resveratrol biosynthesis. The obtained data first demonstrate that overexpression of a CDPK gene resulted in increased accumulation of a stilbenoid phytoalexine in transgenic plant cells. We propose that the VaCPK20 gene could play an important role in the regulation of resveratrol biosynthesis in grape cells.

Structure and expression profiling of a novel calcium-dependent protein kinase gene, CDPK3a, in leaves, stems, grapes, and cell cultures of wild-growing grapevine Vitis amurensis Rupr.

KEY MESSAGE : VaCDPK3a is actively expressed in leaves, stems, inflorescences, and berries of Vitis amurensis and may act as a positive growth regulator, but is not involved in the regulation of resveratrol biosynthesis. Calcium-dependent protein kinases (CDPKs) are known to play important roles in plant development and defense against biotic and abiotic stresses. It has previously been shown that CDPK3a is the predominant CDPK transcript in cell cultures of wild-growing grapevine Vitis amurensis Rupr., which is known to possess high resistance against environmental stresses and to produce resveratrol, a polyphenol with valuable pharmacological effects. In this study, we aimed to define the full cDNA sequence of VaCDPK3a and analyze its organ-specific expression, responses to plant hormones, temperature stress and exogenous NaCl, and the effects of VaCDPK3a overexpression on biomass accumulation and resveratrol content in V. amurensis calli. VaCDPK3a was actively expressed in all analyzed V. amurensis organs and tissues and was not transcriptionally regulated by salt and temperature stresses. The highest VaCDPK3a expression was detected in young leaves and the lowest in stems. A reduction in the VaCDPK3a expression correlated with a lower rate of biomass accumulation and higher resveratrol content in calli of V. amurensis under different growth conditions. Overexpression of the VaCDPK3a gene in the V. amurensis calli significantly increased cell growth for a short period of time but did not have an effect on resveratrol production. Further subculturing of the transformed calli resulted in cell death and a decrease in expression of the endogenous VaCDPK3a. The data suggest that while VaCDPK3a acts as a positive regulator of V. amurensis cell growth, it is not involved in the signaling pathway regulating resveratrol biosynthesis and resistance to salt and temperature stresses.