Genetic Diversity and Structure of Terminalia bellerica (Gaertn. Roxb.) Population in India as Revealed by Genetic Analysis.

In this study, an extensive exploration survey of wild progeny was conducted which yielded 18 candidate plus trees (CPTs) of Terminalia bellerica. Seeds of these CPTs were collected from diverse locations between 10°54' and 28°07' E longitude, and 76°27' and 95°32' N latitude, covering 18 different locations across 5 states of the Indian subcontinent. The objective of the progeny trial was to assess genetic associations and variability in growth and physio-chemical characteristics. Significant variations (p < 0.05) were observed among the growth traits, encompassing plant height, basal diameter, girth at breast height and volume, as well as physio-chemical characteristics such as leaf length, width, area and chlorophyll content, carotenoids, and protein in the progeny trial. Broad-sense heritability (h2b) estimates were consistently high, exceeding 80% for all growth and physiological related traits under investigation except for plant height, leaf length, and girth at breast height. A correlation study revealed that selecting based on plant height, leaf area, and girth at breast height effectively enhances T. bellerica volume. A moderate genetic advance in percent of the mean (GAM) was observed for most traits, except leaf length, leaf width, girth at breast height, and plant height. Across all 13 traits, phenotypic coefficient of variation (PCV) surpassed genotypic coefficient of variation (GCV). Utilizing principal component analysis (PCA) and dendrogram construction categorized the genotypes into seven distinct groups. In conclusion, the study has demonstrated that targeting girth at breast height and plant height would be a highly effective strategy for the establishment of elite seedling nurseries and clonal seed nurseries for varietal and hybridization programs in the future.

Comparative evaluation of the potential anti-spasmodic activity of Piper longum, Piper nigrum, Terminalia bellerica, Terminalia chebula, and Zingiber officinale in experimental animals.

Background: Spasm of muscle is one of the frequent complaints seen by most of the population worldwide. The present study evaluated the efficacy of some of the commonly used herbal extracts against known spasmogens, such as histamine and 5-hydroxytryptamine (5-HT). Material and methods: The study was conducted on isolated guinea pig ileum and rat uterus preparations using histamine and 5-HT, respectively. Five herbal extracts such as Piper longum (P.L), Piper nigrum (P.N), Terminalia bellerica (T.B), Terminalia chebula (T.C), and Zingiber officinale (Z.O) were tested. Herbal extracts at doses 50, 150, 500, 1500, and 5000 mcg/ml were pretreated to the isolated tissue preparation, and the contractile response of histamine and 5-HT was recorded. The efficacy and the inhibitory concentration (IC50) were calculated and statistically analyzed by one-way ANOVA. Results: The study indicated that all five herbal extracts produced a concentration-dependent suppression of histamine and 5-HT-induced responses. A significant (p < 0.05) non-competitive antagonism was observed against the known spasmogen induced smooth muscle contraction for P.L, P.N, T.B, and Z.O in both guinea pigs and rat uterus preparation. Moreover, P.L and P.N completely abolished (100%) the contractile response induced by histamine and 5-HT. Although, T.C produced a concentration-dependent reduction in known spasmogen-induced contraction but the response was found to be statistically non-significant (p greater than 0.05). Conclusion: The finding suggested that P.L. and P.N. have better activity in terms of reducing the spasmogenic contractions compared to other extracts. Additionally, T.B. and Z.O. can lessen the uterine and intestinal contractions brought on by spasmogens. Although P.L and P.N demonstrated better efficacy against the spasmogenic activity of histamine and 5-HT, more research, particularly on isolated phytochemicals of the extracts and involving different experimental models, is required before establishing the precise safety and efficacy against spasmogenic-induced disorders.

Bioinspired Gold Nanoparticle Synthesis Using Terminalia bellerica Fruit Parts and Exploring Their Anti-bacterial Potency In Vitro.

Gold nanoparticles with their excellent biocompatibility are extensively used in pharma and biological applications. Terminalia bellerica (TB) dry fruit parts mediated gold nanoparticles were synthesized using the aqueous extracts. The secondary metabolites screening of the aqueous extracts was done using phytochemical analysis. The green synthesized gold nanoparticles show vibrant colours. They were characterized using UV-Visible spectroscopy, FT-IR spectroscopy, XRD analysis and FE-SEM. The analytical characterization methods ensured the formation of nanoparticles and could predict the nanometric size of the nanoparticles. The study also lay to determine the antibacterial potential of the TB fruit parts and TB fruit parts mediated gold nanoparticles. The pathogens chosen for the study were pathogens from clinical species such as Acinetobacter pneumonia, Bacillus subtilis, and Enterococcus faecalis which cause common infections. The TB fruit part extracts, as well as TB fruit parts mediated gold nanoparticles were capable enough to destroy clinical pathogens. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12088-021-00937-3.

A randomized, double-blind, positive-controlled, prospective, dose-response clinical study to evaluate the efficacy and tolerability of an aqueous extract of Terminalia bellerica in lowering uric acid and creatinine levels in chronic kidney disease subjects with hyperuricemia.

BACKGROUND: Hyperuricemia is an independent risk factor in chronic kidney disease (CKD). Allopurinol and febuxostat are prescription medicines used to treat hyperuricemia but suffer side-effects. Earlier clinical study has shown that an aqueous extract of Terminalia bellerica (TBE), significantly reduced uric acid levels with no serious adverse effects in hyperuricemic subjects. The objective of this study is to determine the efficacy and tolerability of TB in reducing uric acid and creatinine levels in CKD subjects. METHODS: 59-subjects were randomized to three groups-40 mg-once-daily febuxostat, 500 mg-twice-daily and 1000 mg-twice-daily of TBE. Serum uric acid, creatinine levels and estimated-glometular-filtration-rate were measured at baseline, 4, 8, 12, 16, 20, 24-weeks. Biomarkers of oxidative-stress, endothelial function, systemic inflammation, and platelet-aggregation were evaluated at baseline, 4, 8, 12, 24-weeks. Adverse drug reactions were recorded. Statistical analysis evaluated using GraphPadPrism4. RESULTS: 55-subjects completed 24-week study. Starting at 4-weeks, all treatment groups showed a significant decrease in serum uric acid levels from baseline (p ≤ 0.0001). At 24-weeks, febuxostat, T.bellerica 500 mg-twice-daily, and T.bellerica 1000 mg-twice-daily doses decreased mean-percentage serum uric acid by 63.70 ± 4.62, 19.84 ± 6.43 and 33.88% ± 4.95% respectively (p ≤ 0.0001). Significant decrease in serum creatinine with all the groups starting at 16-weeks was seen (p ≤ 0.005-p ≤ 0.0001). At 24-weeks, the mean-percentage change in creatinine levels was 23.71 ± 12.50, 11.70 ± 9.0, and 24.42 ± 8.14, respectively with febuxostat, T.bellerica 500 mg-twice-daily and T.bellerica 1000 mg-twice-daily. Statistically significant (p ≤ 0.05) increase in estimated glomerular filtration rate-(eGFR) was seen at 20 (p ≤ 0.05) and 24-weeks (p ≤ 0.01) for both febuxostat vs T.bellerica 500 mg-twice-daily and T.bellerica 1000 mg-twice-daily vs T.bellerica 500 mg-twice-daily. There was no statistically significant difference between febuxostat and T.bellerica 1000 mg-twice-daily, with an increase of eGFR of 41.38 and 40.39 ml/min/1.73m2 respectively, with the inference that T.bellerica at 1000 mg-twice-daily dose is as good as febuxostat 40 mg-once-daily. Positive improvements were made by all the groups in endothelial function and the related biomarkers and high-sensitivity C-reactive protein. None of the products showed effect on platelet aggregation. CONCLUSION: In this 24-week study Febuxostat 40 mg, T. bellerica 500 mg-twice-daily and 1000 mg-twice-daily, significantly decreased the serum uric acid and creatinine levels, increased eGFR in CKD subjects. T. bellerica 500 mg-twice-daily and 1000 mg-twice-daily were one-third and more than half as effective at 24-weeks, respectively. T. bellerica extract may be considered a natural alternative for reducing serum uric acid levels. TRIAL REGISTRATION: This study was registered with the Clinical Trials Registry - India (CTRI) with the registration number: CTRI/2019/11/022093 [Registered on: 21/11/2019] Trial Registered Retrospectively.

Mass modeling of Belleric Myrobalan and its physical characterization in relation to post-harvest processing and machine designing.

The present research was inducted to assess the physical properties of Terminalia bellerica fruit which is commonly known as belleric myrobalan. The research includes dimensional properties, gravimetric properties, and frictional properties of the T. bellerica. These properties form an important base for the designing of machine or equipment; those are used in the post-harvest operations. The reported average value for the dimensions of the fruit were 26.80 mm, 20.85 mm, 20.85 mm as long dimensional axis, intermediate dimensional axis, and short dimensional axis respectively. The arithmetic mean diameter, equivalent mean diameter, and geometric mean diameter were calculated as 22.54 mm, 22.32 mm, and 22.31 mm respectively. The sphericity was found to be 0.83 while the aspect ratio was 0.78. In the gravimetric properties, true density and the bulk density for the dried fruit of T. bellerica was found to be 0.63 g/cm3 and 0.57 g/cm3 respectively. In the frictional properties, static coefficient of friction was found to be highest on the mild steel and plywood sheet (0.25) and lowest was on the stainless steel sheet (0.19). The angle of repose was found to be 33.96°. Along with the physical properties of the fruit, mass modeling was done for which the fruits were categorized into 3 categories based on their weight. Each category of fruit showed the different significance level and regression factor in every model namely linear, quadratic, power, and S-curve. The quadratic model showed highest level of significance for all the dimensions, area and volume in the small and large mass group while power model was the best fit in the medium mass group. It was concluded that the quadratic model in the small and large mass group while power model in the medium mass group are recommended as best fit and can be used for the design considerations of any post-harvest machine or equipment.

Wound healing activity of Terminalia bellerica Roxb. and gallic acid in experimentally induced diabetic animals.

Background Diabetic wound are the major problem of society because of its delay healing of wound in hyperglycemia patient. The target of this study need to know the gallic acid compelling synergistically wound healing in diabetic animals as it is available in fruit extract or is more effective in given pure form in diabetic animals. Methods The wound healing effects of fruit extract Terminalia bellerica Roxb. and gallic acid were evaluated in experimentally induced hyperglycemic animals. In this investigation, excision and dead space wound models were used. The contraction wound %, re-epithelialization days, area of scar, histopathological examination of tissue, hydroxyproline level and weight of tissue granuloma were evaluated in this study. Results Terminalia bellerica fruit extract (400 mg/Kg) and gallic acid (200 mg/Kg) have been ingested by orally in streptozotocin induced diabetic rats. They were founded significantly (p<0.05) improved percentage wound contraction and decreased the size of scar area and days of re-epithelialization in excision wound model. Also, they were improved the level of hydroxyproline level in diabetic rats as compared to diabetic control group in dead space wound model. Standard drug vitamin C was also founded significantly improved wound healing activity in rats. These results suggested that treated groups accelerated wound healing activity in diabetic rat. Conclusions The observations and results obtained in the present study indicated the wound healing efficacy of fruit ethanolic extract and gallic acid in diabetic animals. Yet, the wound healing effect was increasingly reported with gallic acid.

Grilling enhances antidiarrheal activity of Terminalia bellerica Roxb. fruits.

ETHNOPHARMACOLOGICAL RELEVANCE: Terminalia bellerica Roxb. fruits are rich in a variety of biologically active ingredients. Tharu and Buksa tribes of Udham Singh Nagar, Uttarakhand, India use grilled fruits of Terminalia bellerica as an effective cure for diarrhea AIM OF THE STUDY: We validated the ethnobotanical claim by comparing the antidiarrheal effect of grilled fruits (GF) with dried fruits (DF). MATERIALS AND METHODS: The 50% ethanolic extracts of GF and DF were successively fractionated; the antioxidant and bacterial inhibition activity were studied using DPPH free radical scavenging, anti-lipid peroxidation and broth dilution method respectively. Difference in metabolites of ethyl acetate fractions of GF and DF was analyzed using GC-MS, gallic acid content was determined through HPTLC. Further the in-vivo antidiarrheal effect of ethyl acetate fractions of DF and GF was studied on castor oil induced diarrhea model. RESULTS: The ethyl acetate fractions showed potential DPPH free radical scavenging (IC50 11.13µg/ml in DF and 8.56µg/ml in GF), anti-lipid peroxidation and antibacterial activity. The non-targeted metabolic profiling showed higher content of tartaric acid, valeric acid, gallic acid, succinic acid, oxalic acid, malonic acid, malic acid, 1,2,3 trisbenzene, uridine and 11-eicosenoic acid in GF. The HPTLC results indicated that gallic acid content was 2.8 (±0.14) and 4.92 (±0.28) mg/g while ellagic acid content was 4.7 (±0.32) and 4.45 (±0.45) mg/g dry powder in DF and GF respectively. According to in vivo antidiarrheal activity DF and GF (100mg/kg oral) inhibited diarrhea by 41.87% and 71.72% respectively. CONCLUSION: Grilling significantly altered the levels of metabolites in T. bellerica fruits which could be responsible for its increased therapeutic potential.

A randomized, double-blind, placebo-, and positive-controlled clinical pilot study to evaluate the efficacy and tolerability of standardized aqueous extracts of Terminalia chebula and Terminalia bellerica in subjects with hyperuricemia.

OBJECTIVES: To evaluate the efficacy and tolerability of standardized aqueous extracts of Terminalia chebula and Terminalia bellerica versus febuxostat and placebo on reduction in serum uric acid levels in subjects with hyperuricemia. MATERIALS AND METHODS: A total of 110 eligible subjects with hyperuricemia were enrolled and randomized to either of the five treatment groups - T. chebula 500 mg twice a day (BID), T. bellerica 250 mg BID, T. bellerica 500 mg BID, placebo BID, and febuxostat 40 mg once daily plus an identical placebo - for a duration of 24 weeks. Serum uric acid levels were measured at baseline and at the end of 4, 8, 12, 16, 20, and 24 weeks. Statistical analysis was done using GraphPad Prism Software 4. RESULTS AND INTERPRETATION: All active treatment groups showed a reduction in serum uric acid levels compared to baseline and placebo. Significant reduction in mean serum uric acid levels started as early as 4 weeks following treatment, compared to baseline, with T. bellerica (500 and 250 mg), febuxostat (P<0.001), and T. chebula 500 mg (P<0.01); an increase in serum uric acid levels was seen with placebo (P<0.05). The serum uric acid levels became steady after 16 weeks of treatment and remained the same until the end of 24 weeks. The reduction of serum uric acid levels in the T. bellerica 500 mg group was nearly twice that of the T. chebula 500 mg group as well as T. bellerica 250 mg group at all time points. T. bellerica 500 mg reduced serum uric acid levels from 8.07±0.87 to 5.78±0.25 compared to febuxostat, which reduced serum uric acid levels from 8.53±0.97 to 4.28±0.67 (P<0.001) at the end of 24 weeks. The efficacy of T. bellerica appeared to be dose dependent. All the formulations were well tolerated. CONCLUSION: T. bellerica has the potential for treating hyperuricemia as it was devoid of any serious adverse effects in the present study. Further studies are needed to confirm this potential.

Micropropagtion of Terminalia bellerica from nodal explants of mature tree and assessment of genetic fidelity using ISSR and RAPD markers.

The present study reports an efficient in vitro micropropagation protocol for a medicinally important tree, Terminalia bellerica Roxb. from nodal segments of a 30 years old tree. Nodal segments taken from the mature tree in March-April and cultured on half strength MS medium gave the best shoot bud proliferation response. Combinations of serial transfer technique (ST) and incorporation of antioxidants (AO) [polyvinylpyrrolidone, PVP (50 mg l(-1)) + ascorbic acid (100 mg l(-1)) + citric acid (10 mg l(-1))] in the culture medium aided to minimize browning and improve explant survival during shoot bud induction. Highest multiplication of shoots was achieved on medium supplemented with 6-benzyladenine (BA, 8.8 µM) and α-naphthalene acetic acid (NAA, 2.6 µM) in addition to antioxidants. Shoot elongation was obtained on MS medium containing BA (4.4 µM) + phloroglucinol (PG, 3.9 µM). Elongated shoots were transferred to half strength MS medium containing indole-3-butyric acid (IBA, 2.5 µM) for root development. The acclimatization of plantlets was carried out under greenhouse conditions. The genetic fidelity of the regenerated plants was checked using inter simple sequence repeats (ISSR) and randomly amplified polymorphic DNA (RAPD) analysis. Comparison of the bands among the regenerants and mother plant confirmed true-to-type clonal plants.

Expression of pro-inflammatory interleukin-8 is reduced by ayurvedic decoctions.

Eleven decoctions, obtained from indian plants widely used in ayurvedic medicine, have been investigated as a possible source of molecules exhibiting biological activity on the interaction between DNA and NF-kB, a transcription factor involved in the expression of proinflammatory genes. Cystic fibrosis (CF) cell line stimulated by TNF-α has been used as inflammatory cellular model to determinate interleukin-8 (IL-8), one of the most relevant pro-inflammatory mediator in CF regulated by the NF-kB. The chemical characterization of these 11 decoctions by spectrophotometric analysis and NMR fingerprinting highlighted that sugars and polyphenols seemed to be the main compounds. Our results demonstrated that Azadirachta indica, Terminalia bellerica, Terminalia chebula, Hemidesmus indicus, Emblica officinalis and Swertia chirata are the most active decoctions in inhibiting NF-kB/DNA interactions by EMSA assay and in reducing pro-inflammatory IL- 8 expression in CF cells at IC50 concentrations by Real-Time and Bio-plex analyses. Finally, we observed the increase of all inhibitory activities with the rise of total polyphenols, procyanidins and flavonoids, except for the levels of IL-8 mRNA accumulation, that were as high as flavonoid content grown up by the statistical multivariate analyses. In conclusion, these six decoctions might be interesting to explore new anti-inflammatory treatments for diseases, such as CF.

Immunomodulatory effects of triphala and its individual constituents: a review.

The role of plant extracts and Ayurvedic polyherbal preparations in treating various ailments has been acknowledged since time immemorial. Studies based on the effect of these extracts in treatment of different diseases have also been well documented. Indian medicinal literature also emphasizes the synergistic effect of polyherbal drugs in restoring and rejuvenating immune system. This review focuses on the immunomodulatory potential of the polyherbal preparation, Triphala and its three constituents, Terminalia bellerica, Terminalia chebula and Emblica officinalis. The role of Triphala and its extract has been emphasized in stimulating neutrophil function. Under stress condition such as noise, Triphala significantly prevents elevation of IL-4 levels as well as corrects decreased IL-2 and IFN-γ levels. Under the condition of inflammatory stress its immunosuppressive activity is attributed to its inhibitory action on complement system, humoral immunity, cell mediated immunity and mitogen-induced T-lymphocyte proliferation. The aqueous and alcoholic extracts of the individual constituents reportedly enhance especially the macrophage activation due to their free radical scavenging activity and the ability to neutralize reactive oxygen species. This study thus concludes the use of Triphala and its three individual constituents as potential immunostimulants and/or immunosuppressants further suggests them to be a better alternative for allopathic immunomodulators.

Acute and chronic toxicity studies of the water extract from dried fruits of Terminalia bellerica (Gaertn.) Roxb. In Spargue-Dawley rats.

Acute and chronic toxicities of the water extract from the dried fruits of Terminalia bellerica (Gaertn.) Roxb. were assessed in both female and male rats. For the study of acute toxicity, a single oral administration of the water extract at a dose of 5,000 mg/kg body weight (10 female, 10 male) was performed and the results showed no signs of toxicity such as general behavior changes, morbidity, mortality, changes on gross appearance or histopathological changes of the internal organs of rats. The study of chronic toxicity was determined by oral feeding both female and male rats (10 female, 10 male) daily with the test substance at the dose of 300, 600 and 1,200 mg/kg body weight continuously for 270 days. The examinations of signs of toxicity showed no abnormalities in the test groups compared to the controls. In addition, these rats were analyzed for final body and organ weights, necropsy, as well as hematological, blood chemical and histopathological parameters. Taken together, the water extract from the dried fruits of T. bellerica did not cause acute or chronic toxicities in either female or male rats.

Constituents from Terminalia species increase PPARα and PPARγ levels and stimulate glucose uptake without enhancing adipocyte differentiation.

ETHNOPHARMACOLOGICAL RELEVANCE: The fruits of Terminalia bellerica Roxb. (Combretaceae) and T. chebula Retz. (Combretaceae) are important components of triphala, a popular Ayurvedic formulation, for treating diabetes in Indian traditional medicine. AIM OF THE STUDY: The aim of this study was to evaluate the effects of the constituents of T. bellerica and T. chebula fruit extracts on PPARα and PPARγ signaling/expression, cellular glucose uptake and adipogenesis. MATERIALS AND METHODS: PPARα and PPARγ signaling and expression (luciferase assay and western blot) and the insulin-stimulated uptake of 2-NBDG were determined in HepG2 cells. The effects on adipogenesis were determined in 3T3-L1 cells by Oil red O staining and measurement of lipid content by AdipoRed reagent. RESULTS: Out of the 20 compounds, two ellagitannins, chebulagic acid (1) and corilagin (2), and three gallotannins, 2,3,6-tri-O-galloyl-ß-D-glucose (3), 1,2,3,6-tetra-O-galloyl-ß-D-glucose (4), and 1,2,3,4,6-penta-O-galloyl-ß-D-glucose (5), showed the enhancement of PPARα and/or PPARγ signaling. Two of the gallotannins (4 and 5) also increased PPARα and PPARγ protein expression, while all three (3-5) enhanced insulin-stimulated glucose uptake into HepG2 cells. Compound 1,2,3,6-tetra-O-galloyl-ß-D-glucose (4) was the most potent in increasing cellular glucose uptake (9.92-fold increase at 50 µM). In the test for adipogenesis, 3-5 did not enhance the differentiation of 3T3-L1 preadipocytes but inhibited the adipogenic effect of rosiglitazone. CONCLUSION: Three gallotannins (3-5) from Terminalia fruits acting as enhancers of both PPARα and PPARγ signaling increased insulin-stimulated glucose uptake without inducing the adipogenesis, with 1,2,3,6-tetra-O-galloyl-ß-D-glucose (4) being the most effective in stimulating glucose uptake and 1,2,3,4,6-penta-O-galloyl-ß-D-glucose (5) being most effective in increasing PPAR protein expression.

Quantification of gallic acidin fruits of three medicinal plants.

Triphala is a traditional herbal formulation consisting of dried fruits originating from three medicinal plants, namely Terminalia chebula, Terminalia bellerica and Phyllanthus emblica. It is used in folk medicine for the treatment of headaches, dyspepsia and leucorrhoea. There are some reports regarding Triphala's pharmacological effects including its anti-cancer, radioprotective, hypocholesterolaemic, hepatoprotective and anti-oxidant activities. The most important components of these plants are the tannins and gallic acid which they contain. Gallic acid being a compound with tannin structure existing in the Triphala fruit. In this research, the gallic acid content contained in the three plants constituting Triphala was determined. Plant fruits were purchased from available Iranian markets. Milled and powdered fruits from each plant were extracted with 70% acetone and subjected to a reaction with rhodanine reagent in the process forming a colored complex. The complex's absorbance was measured at 520 nm and the amount of gallic acid was determined using its calibration curve. According to the results, the highest amount of gallic acid was observed in Phyllanthus embelica (1.79-2.18%) and the lowest amount was found in Terminalia chebula (0.28-0.80%). Moreover, differences between plant samples from different markets places were found to be statistically significant (p < 0.05). These differences can possibly be due to the source of plant preparation, storage condition and period of Triphala storage. In general, the rhodanine assay is a simple, rapid and reproducible method for the standardization of Triphala as gallic acid.