Woodfordia fruticosa (L.) Kurz: Integrating Phytochemistry, Novel Bioactivities and Sustainable Utilization Strategies

INTRODUCTION

W. fruticosa (Figure 1) belong to the family of "Lythraceae" (Table 1), commonly known as Dhataki, Dhaava, Pushpa (Table 2), which grow in tropical and subtropical regions. ¹ According to Ayurvedic books, W. fruticosa is one of the most important fermentation agents in Arishtas and asavas. ² Aristas are prepared from plant decoctions, while Asavas are made from plant juices. Aristas are used as cardio tonics and digestives. ³ Arishtas act as general health stimulants. As per CCRIMH monograph (1978), 18 arishtas are mentioned. Out of 18 arishtas, 17 formulations of them include W. fruticosa extract as a key ingredient. ⁴ It has a wide geographical range, occurring across parts of Africa (Tanzania, Madagascar, Comoros), the Middle East (Oman, Saudi Arabia), and Asia (Bhutan, China-Guangdong, Guangxi, Myanmar, Yunnan-India, Sri Lanka, Indonesia, Nepal, Pakistan, and Vietnam). ^5,6,7 In Indonesia and Malaysia, the dried flowers are known as sidowava or sidawayah which are used for improving fertility in women. ⁸ The plant blooms abundantly between January and April, during which its leaves shed and new ones sprout in February-March. The flowers are small, bitter in taste, cold in nature. ⁹ Plant all parts flowers, roots, leaves, stem bark, creepers, flowers and fruits are considered beneficial. They are used in treatment of bone diseases, fever, diarrhea, piles, ulcers, cough, asthma, antiviral, antipyretic, antifertility, granuloma-inhibitory, antitumor, hepatoprotective, immunomodulatory and antihyperglycemic effects. ¹⁰ In Nepal and India its leaves are traditionally used to treat fever when combined with dried ginger and sugar and it also used in the management of leucorrhoea, piles and sitz baths etc. ¹¹ The plant is being overharvested which puts it at risk. At the same time, traditional medicinal uses are not being connected to modern techniques like biotechnology and novel formulations. To integrate new and old knowledge. A fresh review is needed to connect traditional information with recent science, highlight weak spots and effective use in medicines. ^12,13

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Morphological Characteristics

Morphology of W. fruticosa are summarized in Table 3 including habit, seeds, flowers, bark, leaves, shoots.

Phytochemistry

W. fruticosa contains numerous natural compounds. These include tannins, flavonoids and phenolics which are common plant compounds with antioxidant and protective properties (Table 4 and Figure 2). Researchers found that it's flowers is extremely rich in mesi-inositol, hecogenin, chrysophenol, β-sitosterol, octacosanol, diglucoside, gallic acid, methyl gallate, isoschimawalin A, tellimagrandin, heterophylliin A, gemin D, pentagalloylglucose, tetragalloylglucose, kaempferol, caffeic acid, chlorogenic acid, catechin, rutin, butyric acid, furfural, α-terpineol, 4-terpinenol, linalool, farnesol, α-humulene, β-myrcene, pyrazines, safrole, Methyl nonanoate, Eugenol acetate, Nonanoic acid, 2-Acetylpyrrole, pentadecanoic acid, γ-terpinene, dihydrocarvyl acetate, hexadecenoic acid, ellagic acid, myricetin, Nerol, Epoxylinalol, [Z,E]-α-Farnesene, Junipene, Butanoic acid, Benzeneacetaldehyde, Linalool acetate, 5-Methylfurfural, Octanol, 2-Nonenal, 2-Acetylfuran, 2-Ethylhexanol, Tetramethylpyrazine, 2-Methyltetrahydrofuran, sulcatone, 3-Hexenol, pelargonaldehyde, Tetradecane, 2-Pentylfuran, 2-Methylbutanol, 2-Hexenal, 2-Butanone, 3-Methylbutanal, 2-Ethylfuran, 2,3-Butanedione, Pentanal and 4-Terpineol. ^{4,5,9,15,16,17} Leaves are abundant in many compounds such as oleanolic acid, ursolic acid, lupeol, cosmene, γ-curcumene, elemol, β-selinene, germacrene-D, β-caryophyllene, woodfruticosin, glucogallin, quercetin, myricetin3-O-α-L-arabinopyranoside, cyclopropaneoctanoic acid, palmitic anhydride, dihydrocarvyl acetate, 2H-1-benzopyran-2-one, caryophyllene epoxide and lawsone. ^4,9,18,19

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Stem bark contains octacosanol, norbergenin, gum, c-glucosidenorbergenin and identified number of elements using energy spectroscopy such as hydrogen, phosphorus, potassium, calcium, chlorine, iron, carbon, calcium and sulfur were found in leaves while carbon, hydrogen, magnesium, phosphorus, calcium, platinum, sulfur, chlorine, potassium, silicon, and iron. ^4,9,20

Novel Bioactivities

Anti diabetic activity

Aqueous extract of plant lowers the raised glucose levels in blood to confirm this activity researcher was used alloxan to induce diabetes and extract were given orally. The rat who received the plant extract gained weight over time and displayed lower blood sugar levels than the untreated diabetic rats which indicated improved health. For comparison standard drug glibenclamide was used. ²⁸ Dexamethasone increases glucose and triglycerides levels in mice while leaves extracts lowering blood sugar and triglycerides similar to pioglitazone. ²⁹

Antioxidant activity

Methanolic extract of leaves was evaluated using DPPH free radical technique and strong effect with an IC₅₀ value of 1.86 µg/ mL with high potency. ¹¹ W. fruticosa flowers protected rats from carbon tetrachloride which cause liver and kidney damage. This indicates that the plant has a high potential for antioxidants. ^30,31

Hepatoprotective activity

Methanolic extract of flowers affects liver cancer caused by N-Nitrosodiethylamine in rats and extract protected liver by increasing antioxidant enzymes and lowering harmful markers in the blood like ALP, bilirubin, LDH, MDA, AFP and prevent or slow liver cancer progression ^32,33 and flowers protected rat liver from diclofenac induced damage. ³⁴

Wound healing

W. fruticosa gel was prepared to speed up recovery in mice for this activity three groups were compared and flowers gel enhanced healing by lowering inflammatory cells and encouraging tissue repair. ^7,35

Antimicrobial activity

W. fruticosa extract was tested against E. coli, Shigella species show small inhibition zones and Cryptococcus neoformans, Candida albicans and results confirms that extract preventing fungal adhesion and also disrupting biofilm structures. ³⁶ In this study various extract is tested but methanol extract of leaves showing inhibition zones of 5-7 mm against Aspergillus niger, Trichoderma harzianum, Alternaria burnsii, Alternaria solani and Fusarium oxysporum (Fungi) and E. coli, Salmonella typhi, Bacillus subtilis and Staphylococcus aureus (bacteria). ^37,38,39,40

Gastrointestinal Effects

Flowers extract decreased dangerous bacteria like Alistipes, Parabacteroides and Bacteroides while increasing good bacteria like Akkermansia, Lactobacillus and Bifidobacterium. ^41,42

Immunostimulatory activity

Flowers enhances the body nonspecific immunity by stimulating bone marrow cells and release of lysosomal enzymes, nitric oxide, superoxide and myeloperoxidase from macrophages responsible to improves their capacity to engulf and destroy harmful particles. ^43,44

Antipsoriatic activity

Researcher use mice to treat psoriasis by using combination of formaldehyde and complete freund's adjuvant. Result shows that formulations decreased redness, erythema and scaling. ⁴⁵

Antifertility

Alcoholic extract demonstrated strong antifertility effects in female rats by preventing pregnancy. ⁴⁶

Thrombolytic activity

It's activity was tested using clot lysis technique and result showed that aqueous leaves extract breaking down 41.24% of the clot and chloroform (38.80%), methanol extract (38.22%) and methanol extract of bark have highest activity at 44.90%. ⁴⁷

Antidiarrheal activity

Leaves and bark extract of leaves were given to mice. Result shows that bark extract reduced diarrheal feces by about 56% and 63% while leaves extract reduced them about 69% and 29%. ⁴⁸

Antiulcer activity

Aqueous extract of flower demonstrated strong protective effect than the standard antiulcer drug omeprazole. ⁴⁹ W. fruticosa contain tannins which is responsible for antiulcer activity in this study diclofenac sodium is used to induced ulcers in rats and demonstrated strong protective properties. ^50,51

Anti-inflammatory action

Egg protein denaturation method was used to test W. fruticosa leaves anti-inflammatory properties which mimics inflammation caused by protein damage. Results showed that W. fruticosa extract decreased swelling on the paw and knee and TNF-α levels are also decreased and prevent and reduces inflammatory responses in animals. ^52,53

Antiatherosclerosis action

Methanolic extracts were examined to determine atherosclerosis caused by triton and high fat diet. Result decrease HDL and increase triglycerides, cholesterol, harmful lipoproteins and decreased tissue damage. ⁵⁴

Antiasthmatic activity

Ethanolic extract reversed contractions caused by histamine and acetylcholine by causing total bronchodilation at concentration of 1.6-3.2 mg and protect spasms, airway narrowing and also reducing frequency of cough. ⁵⁵

Anti-urolithiasis activity

W. fruticosa extracts demonstrated potent anti urolithiasis properties by preventing kidney damage and stone formation. ⁵⁶

Antidepressant activity

Flower extract was given to animal orally for 14 days and activity were examined using forced swim test and tail suspension test and extract decreased immobility time and shows strong antidepressants activity. ⁵⁷

Antiviral activity

Flower extract demonstrates strong enterovirus 71 with an IC₅₀ of 1.2 μg/mL with no toxicity up to 100 μg/mL. ⁵⁸

Alzheimer Disease

Scopolamine was used to induced memory impairments in the mice and used Elevated Plus Maze and Morris Water Maze technique. Result shows that extract enhanced the mice's memory and learning similar to the effect of standard drug piracetam. ⁵⁹

Anticancer activity

Flowers and leaves extract was evaluated on human liver cancer cells (PLC/PRF/5) and determine cytotoxic effect of human breast (MDA-MB-231) and pancreatic (PANC-1) cancer cells. Result shows it eliminates cancer cells, decreased liver tumours and extracts had an IC₅₀ value 230.5 μg/mL for pancreatic cancer cells and 124 μg/mL for breast cancer cells. ^60,61

Antinutritional Factors

Numerous antinutritional chemicals were found in W. fruticosa flowers and study shows that phytate, tannin and oxalate are present whereas saponins are absent. Oxalates can cause kidney stones, phytates can decrease the absorption of minerals including iron, calcium and zinc and tannins also obstruct the digestion of proteins. However, in W. fruticosa has mild amounts that are safe. ⁶²

Clinical Studies

In this study W. fruticosa powder mixed with honey to an infant's gums during teething was investigated. 35 out of 40 finish the clinical trial. For half a year parents used this drug three times a day along with regular checkups. common teething symptoms like vomiting, fever, coughing and diarrhea were reduced by drug also help in maintaining normal weight gain. It is safe, efficient and reduces problem related to teeth. ⁶³ In this study herbal mouthwashes made from leaves of W. fruticosa and peels of Punica granatum could benefit with chronic periodontitis. Ethanolic extracts are used to prepare mouthwashes. Total 100 patient are divided into two groups in which one group used Woodfordia leaves mouthwash and other used Punica peel mouthwash. Then researchers evaluated gum health and salivary amylase levels and both groups clearly reduced especially after 1 hr of using the mouthwash and shows potent anti-inflammatory effects. ⁶⁴

Herbal Formulations

Tablet W. fruticosa tablet formulation were created by researchers and tablets cleared all parameters such as friability, weight variation, hardness and disintegration time then tablets were tested in vitro for therapeutic properties and they demonstrated good antimicrobial, anti-inflammatory and antiulcer activity. ⁶⁵

Hand sanitizer

Study focused on preparation of non-alcoholic hand sanitizer from flowers of W. fruticosa known as Dhathaki Arka. Alcohol based sanitizers that dry out the skin, this ayurvedic preparation effectively fights with germs and safe for skin. Result reported good sanitization and have antimicrobial activity. ⁶⁶

Vanishing cream

Study focused on creating vanishing herbal cream from W. fruticosa leaves. Numerous characteristics of cream are tested such as spread ability, pH, viscosity, compatibility and washability. Result demonstrate that the cream was smooth and uniform had pH of 5.9 and cream was yellowish green with pleasant smell. It was stable and simple to apply leaving no residue and forming a non-greasy layer with viscosity of 27,025 cps. Test verified that it was an oil in water emulsion which is typically more comfortable to apply topically. ⁶⁷

Sustainable Utilization Strategies

The goal of study was to create a technique for micropropagation of W. fruticosa which is at risk of going extinct because of its extensive use in ayurvedic medicine. Plant growth regulators such as auxins and cytokinins were cultivated in different concentration on MS media for shoot tip explants either separately and in combination. For direct shoot regeneration combined effects of these hormones were examined. MS media shows best result contain BAP at 2.0 µM which give 52 shoots per explant. The highest number of roots were produced in half strength MS medium supplemented with 5.0 µM IBA and 3% sucrose for root growth. After two weeks, the regenerated plantlets had a survival rate of roughly 70% and have been hardened and successfully transferred to the greenhouse. ² Using Agrobacterium tumefaciens researchers created a genetic transformation technique for W. fruticosa. Transformed tissue were selected by treating leaves segment with bacterium that carried marker and reporter genes then growth on nutritional media supplemented with antibiotics and plant hormones. Hygromycin resistant shoots were generated then successfully rooted and demonstrated a transformation efficiency of 44.5% with two to three transgenic shoots per explant. The marker gene had incorporated into plant genome confirmed through PCR, RT-PCR, southern blot analysis and GUS staining. By strengthening its therapeutic properties this protocol offers a means of improving the plant. ⁶⁸ Seed are tiny and easily self sow easily in open, well-drained soil but germination is more difficult in nursery conditions. since regular seedbeds frequently fail to generate seedlings, planting seed on broken bricks is most effective artificial approach. Light, well-drained soil and sunny location are essential for growing in nursery beds. It is best to spread seeds on ground, push them gently into the soil without covering them and water them sparingly so avoid wash them away. If soil start to dry up it is advised to water it once or twice a day. The plant can be cultivated by cutting from hardwood and softwood both of which are regarded as simple methods. ⁶⁹ Vegetative propagation by stem cuttings is therefore the most dependable technique. When branch cuttings are planted in moist, shaded soil which root quickly and start to flower within two years making it simple for farmers to cultivate plants. This decreases dependence on wild harvesting and ensures supply for the pharmaceutical and ayurvedic sectors. The authors stress the importance of tissue culture research, organized farming and cultivation of herbal gardens in order to preserve the species satisfy industrial demand and stop habitat loss. ⁷⁰ During growing season, a balanced 10-10-10 fertilizer can be applied sparingly but it Favours well-draining loamy soil and does not require extensive treatment. Healthy growth is maintained by replanting in early spring every two to three years. The plant can be multiplied by cutting obtained during growing season or by seeds propagation. After flowering pruning helps the plant maintain its shape, promote new development and remove unhealthy branches. ⁷¹

CONCLUSION

W. fruticosa are rich in bioactive components which exhibits broad range biological activities such as antiviral, Wound healing, antioxidants, antimicrobial, anticancer, anti-inflammatory, cardioprotective, Alzheimer and antidiabetic effect but most study still remain limited mechanistic and molecular level research and more precise therapeutic pathways are required. Most studies still limited in vitro and animal studies to determine safety, effectiveness and dosage toxicity evaluation, clinical trials and active biomarker characterization are still necessary. Future studies may focus on mechanism based pharmacological evaluation, isolation of phytochemical and formulation development. The sustainable use of W. fruticosa is equally significant because of its ecological importance and increasing popularity in herbal formulations.

For long term sustainability without losing biodiversity methods including cultivation, conservation agricultural intervention is important.

ACKNOWLEDGEMENT

The authors are grateful to the School of Pharmaceutical Sciences, Bahra University, Solan for the support and assistance in the preparation of this review article.

ABBREVIATIONS

W. fruticosa: Woodfordia fruticosa; ALP: Alkaline Phosphate; AFP: Alpha-Fetoprotein; BAP: 6-Benzylaminopurine; CFA: Complete Freund's Adjuvant; DPPH: 2,2-Diphenyl-1-Picrylhydrazyl; GC-MS: gas Chromatography-Mass Spectrometry; GUS: Beta-Glucuronidase; HDL: high-density lipoprotein; HPLC: High-Performance Liquid Chromatography; HPTLC: High-Performance Thin-Layer Chromatography; IBA: Indole-3-Butyric Acid; IC50: Half Maximal Inhibitory Concentration; LDH: Lactate Dehydrogenase; MDA: Malondialdehyde; MS: Murashige and Skoog; PCR: Polymerase Chain Reaction; RT-PCR: Reverse Transcription PCR; TNF-α: Tumour Necrosis Factor-Alpha; VEGF: Vascular Endothelial Growth Factor; WHO: World Health Organization; CCRIMH: Central Council for Research in Indian Medicine and Homoeopathy; HSP70: Heat Shock Protein 70.

CONFLICT OF INTEREST

The authors declare that there is no conflict of interest.

AUTHOR CONTRIBUTIONS

Manish Kumar: Conceptualization, literature survey and drafting. Shruti Thakur: Conceptual guidance, supervision, critical revision and final approval of manuscript. Vishnu Das, Joohee Pradhan, Isha Sharma: Literature survey, editing, and assistance in manuscript preparation.