Induction and Submerged Cultivation of Valeriana Jatamansi Adventitious Root Cultures for Production of Valerenic Acid and its Derivatives

In vitro adventitious roots were induced from Valeriana jatamansi to assess their quality as an alternative ingredient for extraction of secondary metabolites to meet the demand of phytopharmaceutical industries. A significantly (p ≤ 0.05) high root induction (90 %) was achieved on Schenk and Hildebrandt medium fortified with 9.84 µM indole-3-butyric acid. A maximum root biomass (144.09 ± 11.36 g/L) with high relative growth rate (2.01 ± 0.04) and growth index (13.41) was achieved in submerged cultivation. The total valerenic acid derivatives (1525.14 µg/g DW) and acetoxy valerenic acid (534.91µg/g DW) were significantly high in induced adventitious roots, with notable quantity of hydroxyl valerenic acid (919.57 µg/g DW) that otherwise not quantifiable in parent plant parts. In addition, 0.059% essential oil yield consisting of 24.00% patchouli alcohol was also obtained from induced adventitious roots. The phenolic acid derivatives were also significantly higher in adventitious roots (451.58 µg/g DW) as compared to rhizome (187.79 µg/g DW) and leaves (263.68 µg/g DW) of the parent plant. Notably, a substantial increase in phytochemicals was evident at subsequent culture stages with significantly reduced in vitro cultivation cycle (2 months) as compared to field grown plants (24 months). Conclusively, a comparable metabolic profile of in vitro induced V. jatamansi adventitious roots and considerably shorter growth cycle clearly determines its potential as a feasible source of phytoconstituents.

Liquisolid Orodispersible Tablets of Valeriana officinalis L. Root Extraction: Formulation, Development, and Evaluation

Background: Valerian root extract is used in traditional medicine to treat sleep disorders. According to previous studies, sedative effects are related to the presence of valerenic acid. Formulating orodispersible tablets (ODTs) is an effective and cost-benefit technique for accelerating drugs' therapeutic effects. This study aimed to formulate ODTs of valerian root extract and evaluate their properties. Materials and Methods: Valerian root was extracted by percolation in 70% ethanol. The solubility of valerenic acid was investigated in various liquid carriers. The extract was uniformly dispersed in the selected carrier (Tween 80), then mixed with other excipients and compressed into tablets. Fourteen formulations with different amounts of sodium starch glycolate (SSG, as a super disintegrant) and camphor (as a sublimating agent) were prepared. The physicochemical properties of the ODTs, drug release rates, and microbial limit tests (MLTs) were studied. Results: Both SSG and camphor accelerated tablet disintegration rates, and their composition showed a synergistic effect (P<0.05). The infrared spectroscopy revealed no chemical interaction between formulation components. The MLTs confirmed that a limited number of microbial colonies were grown in the liquid medium, and no pathogen growth occurred in the specific culture media. Conclusion: The results revealed that Valerian ODT's physicochemical properties were significantly improved compared to conventional tablets. The technique can be utilized for other poorly water-soluble pharmaceuticals.

Ameliorative Effects of the Sesquiterpenoid Valerenic Acid on Oxidative Stress Induced in HepG2 Cells after Exposure to the Fungicide Benomyl

Valerenic acid (VA) is a sesquiterpenoid and a phytoconstituent of the plant valerian used for sleeping disorders and anxiety. The frequency of using herbal components as therapeutic nutritional agents has increased lately. Their ability to improve redox homeostasis makes them a valuable approach against harmful xenobiotics. The purpose of this study was to evaluate the putative beneficial role of VA against the redox-perturbating role of the fungicide benomyl in HepG2 human liver cells in terms of oxidative stress in the cellular environment and in endoplasmic reticulum (ER). Benomyl increased cell total oxidant status and reactive oxygen species production and decreased total antioxidant status. The expression of genes coding for antioxidant molecules, namely, heme oxygenase-1, alpha glutathione s-transferase, NF-ĸB, and liver fatty acid binding protein, were decreased due to benomyl. VA ameliorated these effects. Benomyl also increased ER-stress-related molecules such as endoplasmic reticulum to nucleus signaling 1 protein, glucose-regulated protein 78, and caspase-12 levels, and VA acted also as a preventive agent. These results indicate that VA exerts ameliorative effects after benomyl-induced oxidative stress. VA, a widely used nutritional supplement, is a compound with potent antioxidant properties, which are valuable for the protection of cells against xenobiotic-induced oxidative damage.

Constituents of Passiflora incarnata, but Not of Valeriana officinalis, Interact with the Organic Anion Transporting Polypeptides (OATP)2B1 and OATP1A2

Herbal medication used in the treatment of sleep disorders and anxiety often contain extracts of Valeriana officinalis or Passiflora incarnata. Valerenic acid in V. officinalis and apigenin, orientin, and vitexin in P. incarnata are thought to contribute to their therapeutic effect. It was the aim of this study to test whether these constituents of herbal extracts are interacting with the uptake of estrone 3-sulfate, pregnenolone sulfate, and dehydroepiandrosterone sulfate mediated by the uptake transporters organic anion transporting polypeptide 2B1 (OATP2B1) or organic anion transporting polypeptide 1A2 (OATP1A2). Madin-Darby canine kidney cells overexpressing OATP2B1 or OATP1A2 were used to determine the influence of the constituents on the cellular accumulation of the sulfated steroids. Subsequently, competitive counterflow experiments were applied to test whether identified inhibitors are also substrates of the transporters. Valerenic acid only interacted with OATP2B1, whereas apigenin, orientin, and vitexin interacted with OATP2B1 and OATP1A2. Competitive counterflow revealed that orientin is a substrate of both transporters, while apigenin was transported by OATP1A2 and vitexin by OATP2B1. In a next step, commercially available P. incarnata preparations were assessed for their influence on the transporters, revealing inhibition of transporter-mediated estrone 3-sulfate uptake. HPLC-UV-MS analysis confirmed the presence of orientin and vitexin in these preparations, thereby suggesting that these constituents are involved in the interaction. Our data indicate that constituents of P. incarnata may alter the function of OATP2B1 and OATP1A2, which could affect the uptake of other compounds relying on uptake mediated by the transporters.

A Review Focused on Molecular Mechanisms of Anxiolytic Effect of Valerina officinalis L. in Connection with Its Phytochemistry through in vitro/in vivo Studies

: Valeriana officinalis L. (Valerianaceae) is one of the most reputed ancient medicinal plants used in modern phytotherapy and traditional medicine. Its root extract is one of the most effective herbal sedatives and tranquilizers, where the plant is also used for treatment of gastrointestinal spasms. V. officinalis has a complex phytochemistry consisting of the esterified iridoid derivatives known as valepotriates (e.g. valtrate, didrovaltrate, isovalerenic acid), sesquiterpenes (e.g. valerenic acid), flavonoids (e.g. linarin, apigenin), lignans (e.g. pinoresinol, hydroxypinoresinol), alkaloids (e.g. actinidine, valerine), triterpenes (e.g. ursolic acid), monoterpenes (e.g. borneol, bornyl acetate). Among them, valerenic acid is a marker compound for standardization of the root extracts of the plant and has been reported in many in vitro/in vivo studies to be responsible for anxiolytic action of the plant. Although modulation of gammaaminobutyric acid (GABA) receptors has been revealed to be the leading mechanism of the plant based on the existence of valerenic acid, several studies described interaction of valerenic acid with glutamergic receptors. In addition to valerenic acid; isovaleric acid, didrovaltrate, borneol, and some lignans have been also proposed to contribute to anxiolytic effect of the plant. In the current review, the data selectively scrutinized from the in vitro/in vivo studies about identifying anxiolytic molecular mechanisms of V. officinalis is focused.

Contribution to the biological interest of Valeriana officinalis: an update

Valeriana officinalis L. (Caprifoliaceae family) has been traditionally used to treat mild nervous tension and sleep problems. The basis of these activities are mainly attributed to valerenic acid through the modulation of the GABA receptor. Moreover, V. officinalis is claimed to have other biological activities such as cardiovascular benefits, anticancer, antimicrobial and spasmolytic. The current review aims to update the biological and pharmacological studies (in vitro, in vivo and clinical trials) of V. officinalis and its major secondary metabolites in order to guide future research. Databases PubMed, Science Direct and Scopus were used for literature search including original papers written in English and published between 2014 and 2020. There have been identified 33 articles which met inclusion criteria. Most of these works were performed with V. officinalis extracts and only a few papers (in vitro and in vivo studies) evaluated the activity of isolated compounds (valerenic acid and volvalerenal acid K). In vitro studies focused on studying antioxidant and neuroprotective activity. In vivo studies and clinical trials mainly investigated activities on the nervous system (anticonvulsant activity, antidepressant, cognitive problems, anxiety and sleep disorders). Just few studies were focused on other different activities, highlight effects on symptoms of premenstrual and postmenopausal syndromes. Valeriana officinalis continues to be one of the medicinal plants most used by today's society for its therapeutic properties and whose biological and pharmacological activities continue to arouse great scientific interest as evidenced in recent publications. This review shows scientific evidence on traditional uses of V. officinalis on nervous system.

DNA-Based Authentication and Metabolomics Analysis of Medicinal Plants Samples by DNA Barcoding and Ultra-High-Performance Liquid Chromatography/Triple Quadrupole Mass Spectrometry (UHPLC-MS)

There is growing interest for medicinal plants in the world drug market. Particularly, Matricaria recutita L., Valeriana officinalis L., Tilia spp., and Camellia sinensis (L.) Kuntze are some of the most consumed medicinal plants for treatment of minor health problems. Medicinal plants are seen as natural and safe; however, they can cause interactions and produce adverse reactions. Moreover, there is lack of consensus in medicinal plants regulation worldwide. DNA barcoding and UHPLC-MS technique are increasingly used to correctly identify medicinal plants and guarantee their quality and therapeutic safety. We analyzed 33 samples of valerian, linden, tea, and chamomile acquired in pharmacies, supermarkets, and herbal shops by DNA barcoding and UHPLC-MS. DNA barcoding, using matk as a barcode marker, revealed that CH1 sold as Camellia sinensis was Blepharocalyx tweediei, and sample TS2 sold as linden belong to Malvales. On the other hand, UHPLC-MS analysis revealed the presence of bioactive compounds (apigenin-7-glucoside, acetoxy valerenic acid, valerenic acid, epigallocatechin, and tiliroside). However, none of samples met minimum content of these active principles (except for valerenic acid in VF3) according to the European Medicines Agency (EMA) and Real Spanish Pharmacopeia. In conclusion, this study revealed the need to incorporate DNA barcoding and HPLC-MS techniques in quality controls of medicinal plants.

Effect of Valerian Extract on GABRB3 Gene MRNA Expression and Sedation in BALB/C Mice

Background: Valerian, also known as Setwall, Baldrianwurzel, or Phu is an herbaceous perennial plant from Europe and Asia, ubiquitously dispersed in almost all countries with a pH of 6-7. Valeriana officinalis is often used to treat sleep disorders, anxiety, fatigue, seizures, epilepsy, and depression as traditional medicine for 2000 years. The main constituent of Valeriana officinalis is Valerenic acid with hydroxyl and acetone group derivatives. Valerenic acid has anxiolytic, tranquilizing, and sleep-inducing effects that have been demonstrated in both animal studies and clinical trials. Valerenic acid inhibits catabolism-enzyme induced breakdown of Gamma-Aminobutyric Acid (GABA) in the brain resulting in sedation. Gamma-aminobutyric acid is the most important brain inhibitory neurotransmitter with key roles in the regulation and function of the central nervous system. The sedative effect of Valerian extract is facilitated by the GABAA receptor β3 subunit. GABRB3 gene is a ligand-gated ion channel that encodes the GABAA receptor β3 subunit. GABRB3 also functions as a receptor for diazepam and other anesthetic drugs (i.e. phenobarbital). Valerian influences the presynaptic component of GABA-ergic neurons that affect the release of synaptic GABA. Valerian also inhibits GABA reuptake and GABA catabolism by inhibiting the enzyme GABA transaminase. On that basis, this study aims to determine the effect of valerian extract on GABRB3 gene mRNA expression and sedative effect in BALB/c mice. Objective: This is an experimental study using an animal model, with a posttest-only control group design to determine the effect of Valerian extract on GABRB3 gene mRNA expression and sedative effect in BALB/c mice. Methods: Twenty selected BALB/c mice were randomly allocated into four groups; each group consisting of 5 mice. Group A (negative control) was given 5 ml of aqua dest (distilled water). Group B (positive control) was given 0.025 mg/10 g of diazepam. Group C (treatment group 1) was given 2.5 mg/10 g of Valerian extract, and group D (treatment group 2) was given 5 mg/10 g of Valerian extract. The drugs were administered for seven days through a gastric gavage. Rotarod test was performed on the seventh day. A blood sample of 1 ml was taken on the first day before drug administration and after the rotarod test on the seventh day to be analyzed using RT-PCR. Results: GABRB3 gene mRNA expression showed a significant increase in group B, C, and D (p <0.0001). There was a significant difference between group C and D. The examination of motor coordination functions (Rotarod test) showed a significant difference (p <0.05) between group A and group B, between group A and group C, and between group A and group D. There was no significant difference between group B and both group C and D. Conclusions: GABRB3 gene mRNA expression was increased significantly after the administration of Valerian extract. Based on the Rotarod test, Valerian extract and Diazepam had a clinically similar sedation effect. A higher dose of Valerian extract does not yield a higher level of GABRB3 gene mRNA expression nor sedative effects.