Orthodox healers have long used herbal remedies to cure anxiety disorders and provide relief, with no recorded side effects. The medicinal plant Cuscuta reflexa was chosen for anti-anxiety studies based on personal interactions with some of the traditional healers and locals in the collection areas, who say that the plant is used to reduce anxiety symptoms in animals. The present study entitled as “Phytochemical Standardization and Anti-Anxiety (Izterab-e-Nafsani) study of Aftimoon Hindi (Cuscuta reflexa Roxb.) On Animal Model” has been carried out at the department of Ilmul Advia in Regional Research Institute of Unani Medicine (RRIUM), and University of Kashmir-Srinagar. The aerial parts of the plants were studied for their pharmacognostic standardization and anxiolytic activity
Pharmacognostic standardization and preliminary phytochemical analysis
The current study demonstrates numerous Pharmacognostic standardisation findings of the aerial parts of Cuscuta reflexa, with the aim of potential investigational references and plant identification.
➢ The macroscopic analysis of drugs refers to the assessment of drugs based on colour, odour, taste, scale, shape, and other special characteristics such as touch and texture. It is a method of qualitative assessment focused on the examination of entire drug morphological and sensory profiles. Conclusions taken from studies arising from impressions on sense organs are referred to as organoleptic assessment (Kokate, Purohit and Gokhale 2005; Veeresham et al., 2003). For the aerial parts of Cuscuta reflexa, all of these parameters were reported. These will aid in the initial detection of the crude drugs being investigated.
➢ Regulated incineration of crude drugs produces an ash residue made up of inorganic materials (metallic salts and silica). This parameter has a wide range of values, making it an important factor to consider when analysing crude drugs. The ash value immediately detects more direct pollutants like sand or earth. Inorganic variables in crude drugs, such as calcium oxalate, silica, and carbonate content, may also affect “total ash” values. The acid-insoluble ash value is determined after removing certain variables with acid (as they are soluble in hydrochloric acid) (Mukherjee, Kontz and Reinhardt, 2002). Cuscuta reflexa's total ash, acid insoluble ash, and water soluble ash were found to be 7.62, 0.91, and 4.61 percent, respectively.
➢ Different extractive values determine the amount of active constituents in a specified amount of medicinal plant material when extracted with solvents. It's used to evaluate crude drugs that are difficult to test using other approaches. The amount of extractive drug yield to a given solvent is often used to estimate the amount of a particular constituent or group of related constituents in a drug. In certain cases, the amount of drug soluble in a given solvent may be used to determine its purity. The extraction solvent should be capable of dissolving large quantities of the target substances (Mukherjee et al., 2002).
➢ The pH value, which defines the basic nature of the constituents, i.e. acidity or basicity, gives a small hint as to the form of constituents present. The basic range constituents are found in the Cuscuta reflexa.
The identity of the phyto-constituents present was determined through qualitative chemical tests. In phytochemical experiments, the hydroalcholic extract of Cuscuta reflexa was subjected to a variety of chemical tests in order to determine the identity of the constituents present. Chemical tests on the extract revealed the existence of a variety of essential constituents.
➢ The extracts of Cuscuta reflexa on hydroalcholic extract showed the presence of phytosterols, saponins, phenolics components, alkaloids, glycosides, tannins, terpenes and flavonoids.
➢ The Phenolic and flavonoid contents of aerial part of Cuscuta reflexa were found out to be as 166.95 mg/g of Gallic acid equivalent and 78.32 mg/g of Rutin respectively.
Anti-anxiety Studies
Mice of either sex were picked at random and divided into six groups for anti-anxiety testing. The elevated plus maze, light and dark arena, and open field test were used on the first and seventh days of the study to explore anti-anxiety effects. The number of entries and the amount of time spent in each arm were both recorded. The results were compared to those of a control group of healthy people. The current study shows that hydroalcholic extract of aerial sections of Cuscuta reflexa has antianxiety effects by demonstrating a strong anxiolytic effect using behavioural criteria such as EPM, light and dark, and open field tests. The EPM model of animal anxiety is now one of the most commonly used. EPM is focused on rodents' innate aversion to open spaces and exploits the tension between aversion and exploration. In the EPM, neophobia, discovery, and approach/avoidance conflict tend to be present in the elicited behaviour profiles (Bourin et al., 2007). It's known as the "approach avoidance conflict model" because it takes advantage of a rodent's inherent opportunity to discover new environments (approach) and their intrinsic aversion to extremely hazardous open spaces (aversion) (avoidance) (Lach et al., 2018). The most representative measures of anxiolytic behaviour are increases in open arm parameters. The total arm entries tend to represent shifts in anxiety or general behaviour, and time spent on the central platform appears to be linked to decision making and/or risk assessment (Blanco et al., 2009).
First day study: Open arm activity in EPM is sensitive to the anxiolytic effect of diazepam. Diazepam as expected significantly increased the open arm entries (8.548± 0.19**) and the time spent in open arms (194.77± 1.84**sec) as compared to the control group (4.295± 0.17) and (71.66± 5.17sec). A decrease in the time spent in open arm compared to control group (129.33±0.65) indicating anxiolytic activity of diazepam.
In the same order as HACA, the hydroalcholic extract of aerial sections of Cuscuta reflexa increased open arm entries and time spent in open arm relative to the control group (400mg/Kg) (145.21±2.68 *) > HACA (200mg/Kg) (26.16±14.76 ns).
7th day study: After chronic care, each group was assessed on the seventh day of the study. The diazepam community showed an improvement in open arm exploration as compared to the first day 182.087±1.54** and hydroalcholic extract of Cuscuta reflexa (159.23±3.42**) compared with the control group.
The light and dark arena, which is based on rodents' natural aversion to bright light, is another important parameter for evaluating anxiolytic activity. The number of changes between light and dark compartments, as well as the amount of time spent in the light arena, decide anxiety indices (Granziera et al., 2013; Zehravi, Maqbool and Ara, 2021).
First day study: The time spent in the light box arena increased significantly when the hydroalcholic extract of Cuscuta reflexa at 400mg/kg was used in this model of anxiolytic analysis (172±6.01**) compared to the control group (117.518±0.56). Nevertheless, the extract's response was lower than that of the control group (182.33±4.33**)
Based on the anxiety studies it was observed that:
● In the EPM model, the amount of open arm enteries and time spent in open arm were higher when the hydroalcholic extract of Cuscuta reflexa was 400mg/kg.
● In a light/dark arena test, it was discovered that a 400mg/kg hydroalcholic extract of Cuscuta reflexa increased the amount of time spent in the light arena.
The final outcome of this study confirms the presence of antianxiety properties in the Cuscuta reflexa. Flavanoids were discovered in abundance in the fruit. Preliminary phytochemical research also revealed the presence of Flavaonids, tannins, phytosterols, alkaloids, and other compounds that may be responsible for the anxiolytic impact. As a consequence, it is essential to isolate the plant phytoconstituent responsible for the anxiolytic effect, as well as to investigate the mechanism of its antianxiety action. Furthermore, the plant is one-of-a-kind, with just a few phytochemical studies conducted on it. The active moiety responsible for it can be identified and purified using NMR, Mass, and IR for future study.
Critical perspective
Science and technology have contributed significantly to the improvement of humanity's standard of living. On the other hand, modern life stress and its associated hardships and tribulations are to blame for the growth in the occurrence of psychiatric diseases. Anxiety disorders impact 15 to 20% of medical clinic patients, making them the most frequent mental illnesses in the general population. The presence and kind of precipitants, as well as the duration and course of primary anxiety disorders, are all classified. Although anxiety disorders can present with somatic symptoms in the absence of a diagnosable medical condition, a third of medical patients who report anxiety have an organic aetiology for their psychiatric symptoms. Psychopharmacological breakthroughs have flooded the market with specialized drugs. For anxiety, depression, epilepsy, and insomnia, benzodiazepines (diazepam, nitrazepam, lorazepam, and alprazolam, among others) are the most widely prescribed synthetic drugs. Long-term use of benzodiazepines, on the other hand, has a number of negative side effects, including hampering of cognitive function, physical dependency, and tolerance. Benzodiazepines have deleterious effects on the respiratory, digestive, and immunological systems of the body, as well as the potential for addiction, and long-term use of benzodiazepines is generally more harmful. The medicinal plant Cuscuta reflexa is very valuable. C. reflexa is a parasitic weed plant that kills a large number of crop plants each year. Cuscuta reflexa is still known as the "wonder medicinal plant" because various chemical compounds with therapeutic characteristics have been extracted from it. Although Cuscuta reflexa is not well-known in the urban world, it is well-known among rural people in various Indian states. In India and overseas, there are very few research activity in this plant. This plant's many parts are used to treat various ailments. More research on the plant is needed so that more formulations can be offered and employed in the treatment of various ailments. Anti-inflammatory, cytotoxic, antipyretic, hepatoprotective, anticonvulsant, nematicide, anti-androgenic, hypocholestrolemic, antiandrogenic, hemolytic, diuretic, dermatigenic, immunostimulant, antiarthritic, antiasthma, and anticancer activities have all been reported in Aftimoon pharmacological studies. The Unani system of medicine is based on a set of holistic principles that assist a practitioner turn a diagnosis into a treatment plan. For treating a patient, the Unani system of medicine employs a variety of techniques. It adheres to a holistic healing philosophy. As a result, it integrates a variety of therapies for curing not only the sickness but also the patient's general health. Herbal medications, which are made from natural things, are used for internal treatment. Although there is growing evidence of the effectiveness of herbs and their preparations in treating anxiety disorders, translating these findings into effective treatment for patients is hampered by a lack of understanding of the chemical composition of the products, a lack of standardization of these preparations, and a scarcity of well-controlled studies. Psychotherapy, nutrition treatment, or medication can be used alone or in combination to treat abnormal or pathological anxiety that has become unbearable and damaging to one's health. Herbal medicines may have a function in the treatment of anxiety disorders, according to preliminary findings, but more research is needed. However, this study is only the first step in that direction, and more research is needed to confirm Aftimoon Hindi's anti-anxiety (Izterab-e-Nafsani) effect (Cuscuta reflexa Roxb.)