Acute and sub-acute toxicity study reveals no dentrimental effect of formononetin in mice upon repeated i.p. dosing.

AIM: Formononetin is a phytoestrogen which possess different pharmacological activities. The intraperitoneal route permits the identification of target organs involved in toxicity without compromising the molecule's bioavailability. The current study investigated the safety profile of intraperitoneal formononetin in Swiss albino mice. MATERIAL AND METHODS: For acute toxicity study, formononetin administered intraperitoneally to mice at the doses of 5, 50, 100, 150, 200, and 300 mg/kg for 14 days. For the subacute toxicity study, mice were intraperitoneally administered with formononetin (12.5, 25, and 50 mg/kg) daily for 28 days. RESULTS: During the acute study, no deteriorating effect was observed on body weight, food and water intake, no behavioral changes were observed in animals. The lethal dose 50% (LD50) of formononetin was determined to be 103.6 mg/kg of BW, with a no observed adverse effect level (NOAEL) of 50 mg/kg of BW. Mortality was observed in the 300 mg/kg dose group and histopathological changes such as a mild degree of diffuse granular degeneration in the liver but for rest all doses did not have any adverse effect. In subacute study, no signs of adverse effects, mortality, no changes in body weight, food and water intake, and hematological and biochemical parameters were observed. Histopathology of subacute study indicates, formononetin did not have any noxious effect on organs. CONCLUSION: Formononetin shows mortality at acute dose 300 mg/kg and LD50 at 103.6 mg/kg of BW, with a NOAEL of 50 mg/kg of BW, rest all doses for acute and sub-acute are safe when given intraperitoneally.

Pharmacological Features of 18ß-Glycyrrhetinic Acid: A Pentacyclic Triterpenoid of Therapeutic Potential.

Glycyrrhiza glabra L. (belonging to the family Leguminosae), commonly known as Licorice, is a popular medicinal plant that has been used in traditional medicine worldwide for its ethnopharmacological efficacy in treating several ailments. Natural herbal substances with strong biological activity have recently received much attention. The main metabolite of glycyrrhizic acid is 18ß-glycyrrhetinic acid (18ßGA), a pentacyclic triterpene. A major active plant component derived from licorice root, 18ßGA has sparked a lot of attention due to its pharmacological properties. The current review thoroughly examines the literature on 18ßGA, a major active plant component obtained from Glycyrrhiza glabra L. The current work provides insight into the pharmacological activities of 18ßGA and the potential mechanisms of action involved. The plant contains a variety of phytoconstituents such as 18ßGA, which has a variety of biological effects including antiasthmatic, hepatoprotective, anticancer, nephroprotective, antidiabetic, antileishmanial, antiviral, antibacterial, antipsoriasis, antiosteoporosis, antiepileptic, antiarrhythmic, and anti-inflammatory, and is also useful in the management of pulmonary arterial hypertension, antipsychotic-induced hyperprolactinemia, and cerebral ischemia. This review examines research on the pharmacological characteristics of 18ßGA throughout recent decades to demonstrate its therapeutic potential and any gaps that may exist, presenting possibilities for future drug research and development.

A standardized polyherbal preparation POL-6 diminishes alcohol withdrawal anxiety by regulating Gabra1, Gabra2, Gabra3, Gabra4, Gabra5 gene expression of GABAA receptor signaling pathway in rats.

BACKGROUND: Alcohol abuse is a major problem worldwide and it affects people's health and economy. There is a relapse in alcohol intake due to alcohol withdrawal. Alcohol withdrawal anxiety-like behavior is a symptom that appears 6-24 h after the last alcohol ingestion. METHODS: The present study was designed to explore the protective effect of a standardized polyherbal preparation POL-6 in ethanol withdrawal anxiety in Wistar rats. POL-6 was prepared by mixing the dried extracts of six plants Bacopa monnieri, Hypericum perforatum, Centella asiatica, Withania somnifera, Camellia sinesis, and Ocimum sanctum in the proportion 2:1:2:2:1:2 respectively. POL-6 was subjected to phytochemical profiling through LC-MS, HPLC, and HPTLC. The effect of POL-6 on alcohol withdrawal anxiety was tested using a two-bottle choice drinking paradigm model giving animals' free choice between alcohol and water for 15 days. Alcohol was withdrawn on the 16th day and POL-6 (20, 50, and 100 mg/kg, oral), diazepam (2 mg/kg) treatment was given on the withdrawal days. Behavioral parameters were tested using EPM and LDT. On the 18th day blood was collected from the retro-orbital sinus of the rats and alcohol markers ALT, AST, ALP, and GGT were studied. At end of the study, animals were sacrificed and the brain was isolated for exploring the influences of POL-6 on the mRNA expression of GABAA receptor subunits in the amygdala and hippocampus. RESULTS: Phytochemical profiling showed that POL-6 contains major phytoconstituents like withaferin A, quercetin, catechin, rutin, caeffic acid, and ß-sitosterol. In-vivo studies showed that POL-6 possesses an antianxiety effect in alcohol withdrawal. Gene expression studies on the isolated brain tissues showed that POL-6 normalizes the GABAergic transmission in the amygdala and hippocampus of the rats. CONCLUSION: The study concludes that POL-6 may have therapeutic potential for treating ethanol-type dependence.

Bacopa monnieri abrogates alcohol abstinence-induced anxiety-like behavior by regulating biochemical and Gabra1, Gabra4, Gabra5 gene expression of GABAA receptor signaling pathway in rats.

Evidence has revealed a high degree of comorbidity of excessive alcohol intake and abstinence anxiety-like behavior. The ethanolic extracts of Bacopa monnieri (BME) are used in Indian traditional medicines for the management of alcoholic disorders. However, the underlying mechanism(s) associated with the influence of BME on alcohol abstinence-induced anxiety-like behavior have not been adequately addressed. Therefore, the present study was planned to examine the beneficial effects of BME in alcohol abstinence-induced anxiety-like behavior and the underlying mechanism of action subsequent to long-term voluntary drinking of alcohol. For the assessment of the effects of BME, Wistar rats were exposed to voluntary ingestion of 4.5%, 7.5% and 9% v/v alcohol for 15 days. The doses (100, 200, and 500 mg/kg) of BME and diazepam (2 mg/kg) were administered via gavage for three consecutive days in the alcohol abstinence period on the days 16, 17, and 18. The behavioral studies were conducted employing the elevated plus maze test (EPM), and light-dark test on day 18 to determine the effects of BME and diazepam in the ethanol abstinence-induced anxiety-like behavior. Alcohol biomarkers like ALT, AST, ALP, GGT, and MCV were estimated using commercially available kits. The expression of Gabra1, Gabra2, Gabra3, Gabra4, Gabra5 genes of the GABAA receptors subunits in the hippocampus as well as amygdala were also examined by reverse-transcription quantitative polymerase chain reaction. The HPLC analysis demonstrated that BME contained 9.9% bacoside-A as a major component. The results revealed that alcohol abstinence group depicted a reduction in the time spent on the open arms, numbers of entries in the open as well as closed arms in EPM test and similarly decrease in latency to the dark chamber, time spent in light chamber and numbers of transitions in LDT. Further, BME at the doses of 200 mg/kg and 500 mg/kg alleviated anxiety-like behavior which was escalated during alcohol abstinence. However, BME (100 mg/kg) exhibited insignificant protection against alcohol abstinence-induced syndrome. The escalated levels of alcohol-intake biomarkers were also reversed by BME at the dose of 200 mg/kg and 500 mg/kg. The down-regulation of Gabra1, Gabra4, and Gabra5 gene expression following alcohol abstinence were also reversed with a higher dose of BME (200 and 500 mg/kg) treatment. These results show that BME abrogates anxiety-like behavior by modulating alcohol markers and Gabra1, Gabra4, Gabra5 gene expression of GABAA receptor signaling pathway in rats.

Protective effect of Convolvulus pluricaulis against neuroinflammation associated depressive behavior induced by chronic unpredictable mild stress in rat.

Depression is a heterogeneous disorder and has been regarded as an inflammatory disease. The aerial parts of the Convolvulus pluricaulis are used in Indian traditional medicines for the management of nervous disorders. However, the influence of methanolic extract of aerial parts of Convolvulus pluricaulis (CPE) on a chronic animal model of depression has not been investigated yet, and associated biochemical changes are still unclear. Therefore, this study investigates the effects of CPE on a chronic rat model of depression and explores its underlying mechanism of action on neuroinflammation and brain monoamines. The antidepressant-like effect of CPE (25, 50, and 100 mg/kg, p.o.) was depicted using the sucrose preference test and the forced swimming test (FST) while CUMS-induced alteration in the locomotor index was measured using the open field test (OFT) and actophotometer. A consecutive one-week treatment of CPE (50, and 100 mg/kg) or fluoxetine (10 mg/kg, p.o.) treatment significantly increased sucrose preference index, reduced immobility time in the FST, and increased the number of squares crossed, the number of rearing in the OFT and locomotion in the actophotometer in the CUMS-exposed rats. Moreover, elevated levels of pro-inflammatory cytokines IL-1ß, IL-6, TNF-α and liver biomarkers ALT, AST were also significantly reversed by CPE (50, and 100 mg/kg) or fluoxetine administration in the CUMS-exposed rats. Furthermore, a one-week treatment of CPE (50 and 100 mg/kg) or fluoxetine also remarkably restored the serotonin and noradrenaline levels in the hippocampus as well as in the prefrontal cortex of the CUMS-exposed rats. However, CPE (25 mg/kg) exerted insignificant protection against CUMS-induced depressive-like behavior and associated neuroinflammation. Therefore, this study demonstrates that CPE exerted antidepressant-like effect which could be mediated by anti-inflammatory potential, restoring liver biomarkers or monoaminergic responses in the stressed rats.

Effect of Withania somnifera Dunal in ethanol-induced anxiolysis and withdrawal anxiety in rats.

Withania somnifera (WS) or its psychotropic preparation is known to play a critical role in morphine, alcohol and benzodiazepines addiction. This study investigates the role of WS in acute ethanol and withdrawal from chronic ethanol consumption using elevated plus maze paradigm in rats. Acute administration of ethanol (1.5-2 g/kg, ip) triggered anxiolytic effect and withdrawal from prolonged ethanol (9% v/v ethanol, 15 days) consumption elicited enhanced behavioral despair (anxiety). Acute administration of WS (50 mg/kg, oral) potentiated the anxiolytic action of subeffective dose of ethanol (0.5 or 1 g/kg, ip). Moreover, the ethanol withdrawal anxiety was markedly antagonized in dose dependent manner by WS at 200 and 500 mg/kg or higher dose of ethanol (2.5 g/kg). However, co-administration of subeffective doses of WS (50 mg/kg, oral) and ethanol also attenuated withdrawal-induced anxiety due to chronic ethanol (9% v/v ethanol, 15 days) consumption. The results suggest the protective effect of WS in the management of ethanol withdrawal reactions.

Protective effect of Withania somnifera dunal root extract against protracted social isolation induced behavior in rats.

This study investigated the effect of Withania somnifera Dunal (WS) root extract and diazepam in social isolation induced behavior such as anxiety and depression in rats. Rats were isolated for 6 weeks and the assessment of changed behavior were done on elevated plus maze (EPM) and forced swim test (FST). Isolation reared rats spent less time into the open arms on EPM and significantly increased immobility time in FST compared to group housed rats. WS (100, 200 or 500 mg/kg, oral) and diazepam (1 or 2 mg/kg, ip) dose dependently increased the time spent and entries into the open arms on EPM test and showed the anxiolytic activity. Subeffective dose of WS (50 mg/kg, oral) potentiated the anxiolytic action of diazepam (0.5, 1 or 2 mg/kg, ip). WS (100, 200 or 500 mg/kg, oral) also reduced the immobility time in FST, thus showed antidepressant effect in both group housed and social isolates. The investigations support the use of WS as a mood stabilizer in socially isolation behavior in Ayurveda.