Ethyl Acetate Extract Of Bark Research Articles

Phytochemical profiling, antioxidant, and tyrosinase inhibitory potential of the Acacia cyclops trunk bark: in vitro combined with in silico approach.

The aim of this study was to analyze the phytochemical profile of Acacia cyclops trunk bark ethyl acetate extract using LC-tandem mass spectrometry for the first time, along with evaluating its antioxidant and anti-tyrosinase properties. Consequently, we determined the total phenolic and flavonoid contents of the extract under investigation and identified and quantified 19 compounds, including phenolic acids and flavonoids. In addition to assessing their antioxidant potential against DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2'-azino-bis-[3-ethylbenzothiazoline-6-sulfonic] acid) assays, in vitro and in silico studies were conducted to evaluate the tyrosinase inhibitory properties of the A. cyclops extract. The ethyl acetate trunk bark extract exhibited a substantial total phenolic content and demonstrated significant antioxidant activity in terms of free radical scavenging, as well as notable tyrosinase inhibitory action (half-maximal inhibitory concentration [IC50] = 14.08 ± 1.10 μg/mL). The substantial anti-tyrosinase activity of the examined extract was revealed through molecular docking analysis and druglikeness prediction of the main selected compounds. The findings suggest that A. cyclops extract holds promise as a potential treatment for skin hyperpigmentation disorders.

Two tetrahydroxyterpenoids and a flavonoid from Xylocarpus moluccensis M.Roem. and their α-glucosidase inhibitory and antioxidant capacity

Context: Scientific verification of the anti-hyperglycemic potential of Kayu sarampa (Xylocarpus moluccensis), which is popularly used for diabetic medication in North Sulawesi, Indonesia, is strongly important to conduct to avoid inappropriate medication that might cause worse conditions. Aims: To investigate the anti-hyperglycemic potential of X. moluccensis stem bark ethyl acetate extract and isolated compounds through the α-glucosidase enzyme inhibitory and antioxidant capacity determination. Methods: The active compound was isolated from ethyl acetate extract of X. moluccensis stem bark. The structures of those compounds were elucidated by infrared spectroscopy, liquid chromatography–mass spectrometry, and nuclear magnetic resonance spectroscopies. To discover the anti-hyperglycemic potency, α-glucosidase enzyme inhibition assay, antioxidant assay (DPPH and FRAP), and in silico study were performed. Results: Three isolated compounds were obtained and characterized as two tetrahydroxyterpenoids compounds, namely xyloccensin E (1) and ruageanin D (2), and one flavonoid compound, namely 3,5,7,3ʹ,4ʹ-pentahidroxyflavan (catechin) (3). Compounds 1 and 3 inhibited the α-glucosidase enzyme with IC50 values of 118.60 and 55.19 µg/mL, respectively, with a competitive inhibition mechanism. This is also in line with in silico findings, and both compounds showed good binding affinity of α-glucosidase protein, which indicated their anti-hyperglycemic activity potential by inhibiting α-glucosidase enzyme. Moreover, compounds 1 and 3 showed antioxidant capacity through the DPPH method with an IC50 value of 54.69 and 2.87 µg/mL. In addition, 100 µg/mL of compounds 1 and 3 also exhibited antioxidant capacity through the FRAP method with values of 66.35 and 213.82 µmol Fe2+/g, respectively. Conclusions: The X. moluccensis stem bark ethyl acetate extract, and isolated compounds exhibit α-glucosidase enzyme inhibitory activity and antioxidant capacity, which confirms its potency as an alternative medication for hyperglycemia issues.

Chemical and biological investigation of Indigofera ammoxylum (DC.) Polhill. red and white phenotypes through feature-based molecular networking

Chemical investigation of ethyl acetate bark extracts of Indigofera ammoxylum red and white phenotypes led to the bio-guided isolation of four previously undescribed flavonoids, named (2S,3R)-3′,7-dihydroxy-4′,6-dimethoxyflavanol (1), (2S,3R)-6-methoxy-7-hydroxyflavanol (2), 2′,3′,7-trihydroxy-4′,6-dimethoxyisoflavone (7) and 2′,5' -dimethoxy-4′,5,7-trihydroxyisoflavanone (8), along with 14 known compounds (3–6 and 9–18). The previously undescribed structures were characterized based on NMR, HRESIMS, UV and IR data. Published spectroscopic data were used to deduce the structure of the known compounds. Eleven of the 18 isolated metabolites were evaluated for anti-inflammatory activity and cytotoxic activity against human liver carcinoma cells and human colon and colorectal adenocarcinoma cells. All tested compounds showed an anti-inflammatory activity (IC50 NO < 25 μg/mL), and compounds 2 and 3 were more selective than the positive control dexamethasone. Afromorsin (6) showed promising cytotoxic properties against both cancer cell lines (IC50 18.9 and 11.4 μg/mL). Feature-based molecular networking approach applied to bark and leaves extracts of the two phenotypes allowed to detect bioactive analogues, belonging to the families of flavones, isoflavones, flavanones, flavanols and flavonols, and to explore the chemodiversity of the species. The red and white phenotypes have a similar composition, whereas bark and leaves contain specific chemical entities. Finally, this approach highlighted a cluster of potentially bioactive and undescribed metabolites.

Therapeutic potential of natural products and antibiotics against bovine mastitis pathogen of cows and buffaloes.

The present study aims toevaluate the prevalence and antimicrobial sensitivity ofStaphylococcus aureus associated with bovine mastitis toselected antibiotics and plant extracts. Inthe current study, 140milk samples were collected from cows and buffaloes. Among the 140samples, 93samples were positive for sub-clinical mastitis based onthe California Mastitis Test (CMT). Out ofthe total positive samples, 45 were confirmed for S.aureus onaMannitol salt agar media. The antimicrobial susceptibility test revealed that 44.82% ofthe isolates were resistant tocefoxitin (oxacillin) confirming methicillin-resistant S.aureus (MRSA) with ahigher percentage (51.61%) inthe buffalo than inthe cow samples. Furthermore, the PCR assay confirmed the presence ofthemecA gene inall the MRSA isolates. Among the seven tested antibiotics, sulfamethoxazole + trimethoprim showed high efficacy (71.1%) against methicillin-susceptible S.aureus isolates (MSSA). Oxytetracycline and sulfamethoxazole+ trimethoprim showed 20% efficacy against MRSA followed byenrofloxacin (10%). Onthe other hand, the tested samples from Pistacia chinensis revealed that the ethyl acetate extract ofbark showed amaximum zone ofinhibition of21.3mm against MSSA and MRSA isolates at3000μg/disc. Moreover, the methanol extract ofCotoneaster microphyllus formed a12.3mm and 9.1mm zone ofinhibition against the MSSA and MRSA isolates, respectively.

Hepatoprotective and Nephroprotective Effects of Haronga (Harungana madagascariensis) in Combination with Antioxidant Compounds in Albino Spraque Dawley Rats

Aim: The aim of this study was to evaluate the effects of Harungana madagascariensis stem bark ethyl acetate extract with antioxidants on the liver and kidney in rats.&#x0D; Study Design: Experimental.&#x0D; Place and Duration of Study: Department of Veterinary Pharmacology and Toxicology, University of Ibadan (from January-February, 2020).&#x0D; Methodology: This was done using Albino rats (Wistar) grouped into 12 of 5 animals per group for protective studies,ethyl acetate extract (EHM) of the plant at 100mg/kg, 200mg/kg and 500mg/kg in combination with each of Ascorbic acid (200mg/kg), Selenium (1mg/kg) and/or Allopurinol (50mg/kg). Group I-III are control groups, IV-VI EHM/Ascorbic acid, groups VII-IX EHM/Se, and groups X-XII EHM/AL. The combination were administered for 8 days prior acetaminophen (800mg/kg) on day 9. Animals were sacrificed on day 10 and samples were collected for biochemical and histopathological evaluations.&#x0D; Results: The study showed that acetaminophen induced-stress on the liver and kidney causing increase in liver biomarkers such as the aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphate (ALP), creatinine, blood urea, total protein and gamma-glutamyl transferase (GGT). The combination of H. madagasacariensis Ethyl acetate extract with Ascorbic acid, Allopurinol and/or Selenium at different doses causes protective effects by reducing the release of biomarkers and stabilized the elevated enzymes level shown in acetaminophen group. The histopathology of the liver in the acetaminophen group showed moderate portal congestion with mild periportal cellular infiltration and a moderate to severe portal fibrosis, while the kidney showed severe diffuse tubular degeneration and necrosis.&#x0D; Conclusion: The EHM (500 mg/kg and 200 mg/kg) in combination with Ascorbic acid, Allopurinol or Selenium protected the liver and kidney from injury induced by Acetaminophen. The Ethyl acetate extract (EHM) of the plant showed synergy with Ascorbic acid, Selenium and Allopurinol to prevent injury on both organs.

Chemical and Biological Profiling of Three Ferulic Acids Alkyl Esters Isolated from Jatropha pandurifolia (Family: Euphorbiaceae) Stem Bark.

The study's objectives include phytochemical profiling and biological (antioxidant, thrombolytic and cytotoxic) analysis of pure chemicals from Jatropha pandurifolia stem bark ethyl acetate extract. Five different compounds including octacosanyl cis ferulate (1), hexacosyl (E)-ferulate (2) triacontyl ferulate (3), β-sitosterol (4) and stigmasterol (5) are elucidated. Their structures determine through 1HNMR analysis and comparison to published data, while three ferulic acid alkyl esters (1-3) were isolated for the first time from J. pandurifolia. Compounds 1, 2, and 3 all have significant thrombolytic potential with respective values of 68.92% ±1.17 (**P&lt;0.01), 66.56% ±2.35 (**P&lt;0.01) and 70.81%±0.98 (**P&lt;0.01) with comparison to standard streptokinase (73.6%±0.76). When compared to BHT (6.82± 0.99 μg/ml) the IC50 (DPPH assay) values were 16.26±1.07 (**P&lt;0.01), 14.12±1.23 (**P&lt;0.01), and 13.16±1.70 μg/ml (**P&lt;0.01). Comparing the three compounds to the reference vincristine sulphate (LC50: 0.52±0.18 μg/ml), of compound 1 (1.56±0.35 μg/ml) (**P&lt;0.01), compound 2 (1.3±0.78 μg/ml) (**P&lt;0.01) and compound 3 (1.29±0.33 μg/ml) (**P&lt;0.01). The results can therefore be interpreted as a concept of isolated molecules having potential for application in additional pharmaceutical research.

Antioxidant and antidiabetic properties of Garcinia cowa Roxb. extracts from leaves, fruit rind, and stem bark in different solvents

The objective of this investigation was to assess the antioxidant and antidiabetic abilities of Garcinia cowa Roxb., a Thai native plant that has a history of use in traditional medicine in several countries. To achieve this goal, different plant parts, such as the leaves, fruit rind, and stem bark, were subjected to extraction with hexane, ethyl acetate, and 70% ethanol using the maceration method. The Folin-Ciocalteu technique was used to quantify the extracts' total phenolic content (TPC). The antioxidant assays, including DPPH, ABTS, and FRAP, and antidiabetic properties through the alpha-amylase and alpha-glucosidase inhibitory capacities of different extracts from G. cowa were assessed. The results revealed that the stem bark extracted with ethyl acetate displayed the highest level of TPC at 153.68 mg GAE/100 g DW. Compared to the other extracts, the stem bark ethyl acetate extract demonstrated the highest antioxidant activity in DPPH, ABTS, and FRAP assays with values of 436.86, 359.17, and 526.98 µmol TE/100 g DW, respectively. Further examination of the antidiabetic effects of stem bark extract in ethyl acetate exhibited the highest alpha-amylase and alpha-glucosidase inhibitory activities with IC50 values of 12.54, and 8.48 mg/mL, respectively. The findings of this research provide initial indications that G. cowa has both antioxidant and antidiabetic attributes and could be viewed as a potential therapeutic agent for managing diabetes.

Antioxidant and anticancer potential of ethyl acetate extract of bark and flower of Tecoma stans (Linn) and In Silico studies on phytoligands against Bcl 2 and VEGFR2 factors

This study was designed to appraise the antioxidant and anticancer competence of solvent extracts of Tecoma stans (Linn) and analyze the phytoligands interaction against Bcl 2 VEGFR2 through in silico studies. The phytochemical analysis revealed that the ethyl acetate extract contains more number of pharmaceutically valuable phytochemicals than other solvent extracts. Among the various phytochemicals, flavonoid was found as a predominant component, and UV–Vis- spectrophotometer analysis initially confirmed it. Hence, the column chromatogram was performed to purify the flavonoid, and High-performance liquid chromatography (HPLC) was performed. It revealed that the flavonoid enriched fraction by compared with standard flavonoid molecules. About 84.69% and 80.43% of antioxidant activity were found from ethyl acetate extract of bark and flower at the dosage of 80 μg mL−1 with the IC50 value of 47.24 and 43.40 μg mL−1, respectively. In a dose-dependent mode, the ethyl acetate extract of bark and flower showed cytotoxicity against breast cancer cell line MCF 7 (Michigan Cancer Foundation-7) as up to 81.38% and 80.94% of cytotoxicity respectively. Furthermore, the IC50 was found as 208.507 μg mL−1 and 207.38 μg mL−1 for bark and flower extract correspondingly. About 10 medicinal valued flavonoid components were identified from bark (6) and flower (4) ethyl acetate extract through LC-MS analysis. Out of 10 components, the 3,5-O-dicaffeoylquinic acid (ΔG -8.8) and Isorhamnetin-3-O-rutinoside (ΔG -8.3) had the competence to interact with Bcl 2 (B-Cell Lymphoma 2) and VEGFR2 (Vascular Endothelial Growth Factor Receptor 2) respectively with more energy. Hence, these results confirm that the ethyl acetate extract of bark and flower of T. stans has significant medicinal potential and could be used as antioxidant and anticancer agent after some animal performance study.

Exploring Schleichera oleasa crude bark extract for its anti-helminthic potential

Schleichera oleosa (Lour.) Oken or Kusum, belonging to family Sapindaceae is an important medicinal plant reported to possess various pharmacological activities viz. analgesic, antibiotic, anti-dysentery, anti-helminthic, antioxidant and anticancer activities. Scientific studies reporting antihelminthic properties of bark extract of this plant are still lacking, therefore, the present work has been designed to investigate the ethanolic and ethyl acetate extracts of bark for its antihelminthic potential. Antihelminthic evaluation was done using Indian earthworms, Pheretima posthuma by determining time of paralysis and death. Phytochemical screening of the extracts revealed the presence of flavonoids, tannins and phenolic acids. The anti-helminthic activity of both extracts (0.1%) was found better than the standard drug Albendazole. Since, both extracts showed promising results, they can be further helpful in designing herbal formulations for commercial use.dry-powder inhaler (DPI) carries medication to lungs as a dry powder, useful against respiratory diseases.

BIOACTIVE CONSTITUENTS FROM BARK OF PELTOPHORUM PTEROCARPUM BAKER EX K. HEYNE

In vitro antimicrobial and antioxidant activities of sequential bark extracts of Peltophorumpterocarpum Baker ex K. Heyne,was tested against four Gram negative, two Gram-positive bacteria and fivefungal isolates. Antioxidant profile was investigated using 2, 2-Diphenyl-1-picryl-hydrazyl and reductioncapability by ferric ion reducing antioxidant potentials method. Bark ethyl acetate extract showed significantactivity against B. subtilis and P. aruginosa (IZ 18.33 ± 0.33 mm and 18.33 ± 0.57 mm with MIC of 62.5 and31.25 μg/ml, respectively) similarly, higher antioxidant potentials with lower IC50value (5 μg/ml; %inhibition of 97.08 at 80 μg/ml), whereas in FRAP method, methanol extract (1463.33 ± 3.33 AAE/mg dwantioxidant potential) show better activity. Further, GC-MS analysis of the ethyl acetate extract confirmed33 compounds. The main identified constituent was 1,2-benzenedicarboxylic acid, bis(2-ethylhexyl) esterwith 48.9% availability.

Isolation and Identification of two Triterpenoids from Ethyl acetate extract of Bark of Boehmeria rugulosa

Column chromatography of purified ethyl acetate extract of bark part of Boehmeria rugulosa afforded triterpenoids (3-oxo-20-demethylisoaleuritolic-28,29-dioic acid and 3-oxo-20-demethylisoaleuritolic-28,30-dioic acid.). Structure of these compounds was elucidated using spectroscopic techniques. This is the first report of isolation of this compound from bark of B. rugulosa.

English

BACKGROUND Stereospermum colais also known as yellow snake tree is widely utilised to alleviate rheumatic pain and inflammation in the conventional medicinal system. Lapachol has been reported to be anti-inflammatory by inhibiting the production of nitric oxide (NO) and tumour necrosis factor (TNF)-alpha by means of modulating the metabolism of arachidonic acid, activation of NF-aB, suppression of nitric oxide synthase (iNOS) expression and expression of cyclooxygenase-2 (COX-2). It has also been analysed for anticancer and antioxidant activity, renal disorders, endometriosis and cardiac dysfunction. The present study investigates the anti– arthritic activity of the ethyl acetate extracts of Stereospermum colais. METHODS The fruit rind of Garcinia indica was used to prepare extract and was quantified using liquid chromatography–mass spectrometry (LC-MS) / MS. Ethyl acetate extract showed increased content of the phytochemical constituent necessary for the treatment of arthritic pain. So, the ethyl acetate extract of bark of S. colais was evaluated for anti-arthritic activity by complete Freund's adjuvant (CFA). Arthritis index, body weight changes, and the biochemical analysis parameters were measured. Histopathological evaluation along with TNF-alpha and interleukin (IL)-6 assays were also studied. RESULTS The ethyl acetate extract showed significant reduction in arthritis index (P &lt; 0.01), paw swelling (P &lt; 0.01) and arthritic score (P &lt; 0.01), thereby demonstrating antiinflammatory potential. A good improvement in the biochemical parameters in extract treated animals indicates good protection against the inflammation. CONCLUSIONS The results show that Stereospermum colais can be used as a potential anti arthritic drug. KEY WORDS Rheumatoid Arthritis, Complete Freund’s Adjuvant, Stereospermum colais, Ethyl Acetate, Anti-Arthritis, Meloxicam

Effects of solvent-solvent fractionation on the total terpenoid content and in vitro anti-inflammatory activity of Serevenia buxifolia bark extract.

Severinia buxifolia (Rutaceae) is often used as a traditional medical plant. The present study was carried out to estimate the effects of solvents (petroleum ether and hexane: ethyl acetate) used in liquid–liquid extraction to total terpenoid content (TTC) and in vitro anti‐inflammatory activity of the extracts obtained from S. buxifolia bark. The results showed that solvent fractionation increased the TTC compared with crude extracts. The hexane: ethyl acetate bark extract fraction (HEF) had the highest TTC (731.48 µg/ml) in comparison with the petroleum ether bark extract fraction (PEF) (564.81 µg/ml) and the crude extract (CE) (184.26 µg/ml). In addition, one of composition of terpenoid of S. buxifolia, namely ursolic acid, was determined by HPLC method from the crude CE and the fractions PEF and HEF: 2.44 μg/g DW, 3.56 μg/g DW and 5.04 μg/g DW, respectively. The samples had an in vitro anti‐inflammatory activity comparable with that of two reference standards (aspirin and indomethacin). Particularly, the HEF fraction had the highest in vitro anti‐inflammatory activity (i.e., albumin denaturation: IC50 = 147.91 μg/mL, heat‐induced hemolysis: IC50 = 159.91 μg/mL, proteinase inhibition: IC50 = 117.72 μg/mL, and lipoxygenase activity: IC50 = 90.45 μg/mL). Besides, the preliminary experiments of this study were conducted to determine the influences of maceration factors (solvent type, temperature, and time) for S. buxifolia bark extract. The TTC ranged from 453.70 to 842.59 mg linalool/g DW, and the extraction yield from 2.40% to 5.120% in all extracts. Based on TTC and EY, the hexane: acetone mixture is recommended as the optimal solvent to obtain the crude bark extract (CE) at 46°C for 24 hr of maceration. Extracts of S. buxifolia bark are a promising source for the treatment of inflammatory diseases.

Assessment of Anti-inflammatory Activity of 3-Acetylmyricadiol in LPSStimulated Raw 264.7 Macrophages.

Pentacyclic triterpenoids are a biologically active class of phytoconstituents with diverse pharmacological activities, including anti-inflammatory action. In the current study, we isolated 3-Acetylmyricadiol, a pentacyclic triterpenoid, from the ethyl acetate bark extract of Myrica esculenta and evaluated it for anti-inflammatory potential. The ethyl acetate bark extract of the M. esculenta was subjected to column chromatography to isolate 3-Acetylmyricadiol. MTT assay was performed to check cell viability. The production of proinflammatory mediators like nitric oxide, IL-6, TNF-α were observed after the administration of 5, 10, 20 μM of 3-Acetylmyricadiol in LPS-activated raw 246.7 macrophages by the reported methods. MTT assay indicated more than 90% cell viability up to 20 μM of 3-Acetylmyricadiol. The administration of 3-Acetylmyricadiol inhibited the production of nitric oxide, IL-6, TNF-α in a dose-dependent manner significantly in comparison to LPS treated cells. The maximum effect was observed at 20 μM of 3-Acetylmyricadiol which resulted in 52.37, 63.10, and 55.37 % inhibition of nitric oxide, IL-6, and TNF-α, respectively. Our study demonstrated the anti-inflammatory action of 3-Acetylmyricadiol and can serve as a potential candidate in the development of the clinically efficient anti-inflammatory molecule.

ISOLATION OF BIOLOGICALLY ACTIVE PLANT SECONDARY METABOLITES IN ALBIZIA CHEVALIERI USING COLUMN-CHROMATOGRAPHIC TECHNIQUES.

Plants have served human beings as natural source for treatments and therapies from ancient times to date; amongst them medicinal herbs which have gain attention because of its wide use and less side effects. Chromatographic techniques have significant role in natural products chemistry as well as contribute dramatically in the discovery of novel and innovative compounds of pharmaceutical and biomedical limportance. In the present study, phytochemical analysis of Albizia chevalieri was carried out for the three Arial parts of the plant extracted with three different solvents (methanol, ethyl acetate and n-hexane). This study also focused on step-by-step visual demonstration of fractionation and isolation of biologically active plant secondary metabolites in A.chevalieri using column-chromatographic techniques. Isolation of bioactive compounds using column-chromatographic involves: Preparation of sample; Packing of column; Pouring of sample into the column; Elution of fractions; and Analysis of each fractions using thin layer chromatography. Based-on the column-chromatographic technique, it was observed that in each fraction one band were detected. Qualitative analysis showed that methanol extracted almost all the secondary metabolites in all the three Arial parts of the plant. After quantification, it was observed that methanol Root extract was having the highest phenolic contents (44.83µg/GAE/g), followed by methanol leaf extract (38.89 µg/GAE/g), and methanol stem bark extract (22.00 µg/GAE/g). Root ethyl acetate extract (8.19 µg GAE/g), stem bark ethyl acetate extract (13.49 µg GAE/g), and leaves ethyl acetate extract (23.66 µg GAE/g), while n-hexane extracts were having a very low phenolic content with; n- hexane root (0.66 µg GAE/g), n-hexane stem bark extract (2.69 µg GAE/g), and n-hexane leaves extract (7.80 µg GAE/g). The presence of high amount of phytochemical compounds suggest that A. chevalieri plant has high medicinal value and can be carefully studied to extract the natural compounds which are beneficial to human beings and that could be commercialized for higher production than using synthetic drugs with side effects. However, depending on nature of research, compounds can be further purified using high performance liquid chromatography (HPLC), and nuclear magnetic resonance (NMR)spectral analyses.

Phytochemical and Biological Studies on Bark Extract of Aporosa wallichii Hook.f.

The ethyl acetate extract of bark of Aporosa wallichii Hook.f. was extensively investigated for the chemical characterization of secondary metabolites and for biological activities. After repeated chromatographic separation and purification, two compounds were elucidated as a phenolic compound ferulic acid (1) and an uncommon triterpene glut-5(6)-en-3β-ol (2) from the ethyl acetate extract of barks of A. wallichii by using high field NMR analyses. This is the first report of isolation of both compounds from this plant species. No significant antimicrobial activity was observed for any fraction after Kupchan partitioning of the extract. During the antioxidant activity assay, the ethyl acetate soluble fraction of A. wallichii demonstrated significant DPPH radical scavenging capacity with an IC50 value of 1.25 μg/mL. The other fractions viz. petroleum ether, aqueous and chloroform soluble fractions exhibited moderate, mild, and weak activity respectively. The petroleum ether soluble fraction demonstrated maximum thrombolytic property (51.33%) which was similar to that of standard streptokinase (66.81%).&#x0D; Dhaka Univ. J. Pharm. Sci. 19(2): 139-143, 2020 (December)

Selective cytotoxic and anti-metastatic activity in DU-145 prostate cancer cells induced by Annona muricata L. bark extract and phytochemical, annonacin

BackgroundAnnona muricata L. was identified as a popular medicinal plant in treatment regimens among cancer patients in Jamaica by a previously conducted structured questionnaire. Ethnomedically used plant parts, were examined in this study against human prostate cancer cells for the first time and mechanisms of action elucidated for the most potent of them, along with the active phytochemical, annonacin.MethodsNine extracts of varying polarity from the leaves and bark of A. muricata were assessed initially for cytotoxicity using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay on PC-3 prostate cancer cells and the ethyl acetate bark (EAB) extract was identified as the most potent. EAB extract was then standardized for annonacin content using High-performance Liquid Chromatography - Mass Spectrometry (HPLC-MS) and shown to be effective against a second prostate cancer cell line (DU-145) also. The mode of cell death in DU-145 cells were assessed via several apoptotic assays including induction of increased reactive oxygen species (ROS) production, reduction of mitochondrial membrane potential, activation of caspases and annexin V externalization combined with morphological observations using confocal microscopy. In addition, the potential to prevent metastasis was examined via inhibition of cell migration, vascular endothelial growth factor (VEGF) and angiogenesis using the chorioallantoic membrane assay (CAM).ResultsAnnonacin and EAB extract displayed selective and potent cytotoxicity against the DU-145 prostate carcinoma cells with IC50 values of 0.1 ± 0.07 μM and 55.501 ± 0.55 μg/mL respectively, without impacting RWPE-1 normal prostate cells, in stark contrast to chemotherapeutic docetaxel which lacked such selectivity. Docetaxel’s impact on the cancerous DU-145 was improved by 50% when used in combination with EAB extract. Insignificant levels of intracellular ROS content, depolarization of mitochondrial membrane, Caspase 3/7 activation, annexin V content, along with stained morphological evaluations, pointed to a non-apoptotic mode of cell death. The extract at 50 μg/mL deterred cell migration in the wound-healing assay, while inhibition of angiogenesis was displayed in the CAM and VEGF inhibition assays for both EAB (100 μg /mL) and annonacin (0.5 μM).ConclusionsTaken together, the standardized EAB extract and annonacin appear to induce selective and potent cell death via a necrotic pathway in DU-145 cells, while also preventing cell migration and angiogenesis, which warrant further examinations for mechanistic insights and validity in-vivo.

Larvicidal activity of some selected medicinal plant extracts against the vector of filariasis

The present study assessed the role of larvicidal activities of hexane, ethyl acetate and methanol dried leaf and bark extracts of Ocimum gratissimum, Gleditsia triacanthos, Eucalyptus sglobulus and Azadirachta aindica against the fourth instar larvae of filariasis vector, Culex quinquefasciatus mosquito (Diptera: Culicidae). Larvicidal activities of four medicinal plant extracts were studied in the range of 4.69 to 1000 mg/l in the laboratory bioassays against early 4th instar larvae of Culex quinquefasciatus. The mortality data were subjected to probit analysis to determine the lethal concentrations (LC50 and LC90) to kill 50 and 90 per cent of the treated larvae of the respective species. All plant extracts showed moderate effects after 24 h of exposure; however, the highest toxic effect of leaf methanol extract of Ocimum gratissimum, bark ethyl acetate extract of Eucalyptus globulus, methanol extract of Azadirachta indicia and methanol bark extract of Gleditsia triacanthos against the larvae of Culex quinquefasciatus with LC50=43.00, 41.02, 40.12 and 20.36 mg/l; LC90=278.32, 218.72, 215.01 and 86.29 mg/l respectively. The result of findings shows that leaf and bark extract of G. tricantha, A .indica, O. gratissimum and E .globulus can be developed as ecofriendly larvicides.

Antistaphylococcal and biofilm inhibitory activities of Frangula alnus bark ethyl-acetate extract

Staphylococcus aureus is one of the leading causative of numerous infections that is resistant to various antibiotics. Since that we are facing with a serious problem of bacterial resistance it is necessary to find a new antibacterial agents in fight with it. Frangula alnus is traditionally used plant but its antibiofilm potential is poorly investigated. The aim of the study was to investigate antibiofilm activity of F. alnus ethyl-acetate extract (FA) toward S. aureus ATCC strains and clinical isolates as well as its influence on respiration in planktonic and biofilm form. The qualitative GCxGC–MS and quantitative LC–MS/MS analysis revealed that FA extract was rich in phenols and flavonoids and emodin, chatechin, and ester 4-ethoxy benzoic acid were the most dominant components. Results obtained through microdillution assay showed that FA possesses strong antibacterial activity. Furthermore, crystal violet staining of biofilm biomass demonstrated that extract had strong effect on biofilm formation of all tested strains while effect on preformed biofilms was less pronounced. The effect on biofilm was confirmed with scanning electron microscopy where the changes in biofilm structure were noticed. The activity of extract on the consumption of O2 and production of CO2 was monitored using the Micro-Oxymax respirometer. Interestingly, respiration of the most strains was decreased in planktonic form as well as in biofilms. Results obtained in this study are a good basis for further research in order to discover the mechanism of action of the FA extract on connection between biofilm and respiration.

In vitro anti-hyperglycemic, antioxidant activities and intestinal glucose uptake evaluation of Endiandra kingiana extracts

Abstract Optimal control of postprandial hyperglycemia is essentially important in the management of diabetes mellitus. The present investigation was undertaken to evaluate the in vitro anti-hyperglycemic, antioxidant properties and intestinal glucose uptake inhibition of Endiandra kingiana (E. kingiana) extracts. Previously, our group has identified and characterized the bioactive compounds of E. kingiana extracts, which had discovered several polyketides; endiandric acids and kingianins. Here, the inhibitory potential of bark-ethyl acetate (BEA), bark-methanol (BM), leaf-ethyl acetate (LEA), and leaf-methanol (LM) extracts of E. kingiana against carbohydrate-hydrolyzing enzymes with their mode of inhibitions were evaluated. Further, the antioxidant activities and inhibitory potential on glucose uptake in Caco-2 human intestinal cell monolayers were determined. Our finding showed that BEA extract exhibited the most potent inhibition activities against α-amylase (IC50 = 2.32 μg/mL) and α-glucosidase (IC50 = 1.83 μg/mL) by following inhibition mode of competitive and non-competitive manners, respectively. Meanwhile, BM extract exhibited notable antioxidant capabilities, as evidenced by strongest free radical scavenging (IC50 = 1.18 μg/mL) and reducing power effect (118.53 mM Fe2+ equivalent/g extract) in couple with highest total phenolic contents (10.17 mg GAE/g extract) compared to other extracts. Mechanistically, both BEA and BM extracts of E. kingiana significantly inhibited glucose uptake in Caco-2 cell monolayers under sodium-dependent condition. Collectively, these findings suggest that BM and BEA extracts of E. kingiana exert in vitro anti-hyperglycemic and antioxidant properties, which can be further utilized as a potential candidate for treatment of hyperglycemia–induced oxidative stress conditions.