Unveiling Annona Reticulata's Bioactive Arsenal for Enhanced Antibiotic Effects.

OBJECTIVE: To study the antibacterial and phytochemical activities of bioactive elements in the leaves of Annona reticulata Linn, a historically used Bangladeshi medicinal plant. METHODS: Shade-dried and crushed plant leaves were soaked with various solvents to obtain samples for different chemical analyses. All extracts were selected for antimicrobial, physicochemical, and Pharmacological investigations. The antimicrobial activity was evaluated using disc diffusion assay, and broth microdilution methods determined potentiation of the activities of the antibiotic antibacterial activity of the plant extracts was investigated using either gram-positive or gram-negative pathogenic wild-type bacteria. RESULTS: From the initial phytochemical and pharmacological studies, it was clear that all extracts, methanol, chloroform, and ethyl acetate, of the leaves of A. reticulata, were proven to process potent bioactive constituents. While differential antimicrobial properties were found to be possessed by all extracts, methanolic extract was the most potent one against all tested microorganisms. It also has potentiated the activities of antibiotics in E. coli. CONCLUSION: Bioactive constituents in the plant extracts were shown to possess phytochemical and antimicrobial activities. More investigation is needed to segregate the chemical components responsible for the respective phytochemical and antimicrobial activities.

Immunomodulatory activity of ethanol extract of Annona reticulata L. leaf in cultured immune cells and in Swiss albino mice.

BACKGROUND: Annona reticulata Linn, has been shown to possess antipyretic, antihelmintic, hypoglycemic, antiulcer and wound healing properties. However, its immunomodulatory role is yet to be explored. OBJECTIVE(S): In the present study, we intended to investigate the effects of A. reticulata leaf ethanol extract on various components of the immune system. MATERIAL AND METHODS: The effects of A. reticulata leaf extract on human peripheral blood mononuclear cells, monocyte (THP1), and human macrophage (U937) cell lines were investigated. An animal study was conducted to observe the effect of the extract on humoral as well as cell mediated immunity. RESULTS: The extract stimulated proliferation of human PBMC, monocytes (THP1), and macrophages (U937) significantly in a dose dependent manner; expression of transforming growth factor-beta (TGF-ß) increased in western blot analysis. Additionally, the extract treated macrophages exhibited features of activation under the microscope with a significant hike in the NO production. Flow cytometry of extract treated human PBMC revealed increased proliferation of lymphocytes (CD4, CD8 & B-cells) along with enhanced intracellular expression of IL-2, IL-6. Animal study data indicate a significant rise in the antibody titer as well as a strong delayed type hypersensitivity response in the extract (150 mg/kg and 300 mg/kg) treated mice; furthermore, the expression of IL-2 and IL-6 in mice PBMC was augmented. CONCLUSION: The collective data evince the immunomodulatory potential of A. reticulata L. leaf.

Susceptibility of Culex quinquefasciatus (Say) Larvae to Methanolic Extracts of Annona reticulata.

<b>Background and Objective:</b> Environment pollution and resistance of many pests to the most frequently used chemical insecticides gave the rationale of altering to replace them with natural herbal extracts for pests and vector control. This study investigated the effect of methanolic extract of <i>Annona reticulata</i> on the mortality and development of larvae of <i>Culex quinquefasciatus</i>. <b>Materials and Methods:</b> Methanolic herbal extracts were analyzed using Reverse-Phase High-Performance Liquid Chromatography (RP-HPLC) to identify the phytochemical compounds in them. Ten mosquito larvae were used as replicates and exposed to each of the five concentrations of the plant extract (30, 100, 150, 200 and 250 mg mL¹) and ten larvae were exposed to double distilled water and considered as control. <b>Results:</b> Phytochemical analysis revealed the presence of phenols, steroids, quinones, tannins and saponins. Statistical analysis showed a significant strong correlation and regression between exposure to the different concentrations of the extract and mortality of <i>Culex quinquefasciatus</i> larvae where R² = 0.982, the Correlation value is 0.991099 (p<0.05). Results also showed that the extracts affect the development of larvae more than mortality. The effect of the extracts on the early larval stages was significantly high compared to the late stages of larvae. <b>Conclusion:</b> According to our knowledge results of this study has been reported for the first time in Saudi Arabia where <i>Annona reticulata</i> is neglected regionally and no study approved its efficacy as a botanical herbal extract against mosquitoes.

Unveiling antioxidant and anti-cancer potentials of characterized Annona reticulata leaf extract in 1,2-dimethylhydrazine-induced colorectal cancer in Wistar rats.

BACKGROUND: Colorectal cancer (CC) is the third most common cancer in the world. Annona reticulata (AR) also known as bullock's heart, is a traditional herb. AR leaf extract was initially investigated for its anti-bacterial, anti-inflammatory, anti-malarial, anti-helminthic, anti-stress, and wound healing properties. Only a few in vitro cancer studies have been conducted on AR. Although few studies have linked AR leaf extract to many cancers, comprehensive studies addressing regulation, biological functions, and molecular mechanisms leading to CC pathogenesis are clearly lacking. OBJECTIVES: The present study aimed to explore the antioxidant and anti-cancer potentials of AR leaf extract in CC. MATERIALS AND METHODS: The MTT assay was used to test the anti-proliferative activity of AR leaf extract in vitro on the HCT116 cell line. Qualitative and quantitative phytochemical characterization was carried out using gas chromatography: mass spectrometry (GC-MS). 1,2-dimethylhydrazine (DMH) was used to establish CC model in female Wistar rats. The acute toxicity of AR leaf extract was tested in accordance with OECD guidelines. Aberrant Crypt Foci (ACF) count, organ index, and hematological estimations were used to screen for in vivo anti-cancer potential. The antioxidant activity of colon homogenate was determined. RESULTS: The alcoholic leaf extract (IC50, 0.55 µg/ml) was found to be more potent than the aqueous extract. Using GC-MS, a total of 108 compounds were quantified in the alcoholic leaf extract. The LD 50 value was found to be safe at a dose of 98.11 mg/kg of body weight. AR alcoholic leaf extract significantly (p < 0.05) decreased ACF count and normalized colon length/weight ratio. AR leaf extract increased RBC, hemoglobin and platelets levels. The AR alcoholic leaf extract reduced the DMH-induced tumors and significantly (p < 0.05) increased the activity of endogenous antioxidant enzymes such as catalase, reduced glutathione, superoxide dismutase, and decreased the lipid peroxidase activity. AR leaf extract reduced the inflammation caused by DMH and helped to repair the colon's damaged muscle layers. CONCLUSION: Based on the findings from the present study, it can be concluded that the alcoholic leaf extract of AR has antioxidant and anti-proliferative properties and can aid in the prevention of CC development and dysplasia caused by DMH.

Ameliorative effect of Annona reticulata L. leaf extract on antihyperglycemic activity and its hepato-renal protective potential in streptozotocin induced diabetic rats.

BACKGROUND: Annona reticulata L. is a traditionally important plant due to its versatile source of medicine and industrial products. It is used to treat cardiac problems, wound healing, diabetes, ulcers and bacterial infections. As it is a commercial fruit bearing plant, wide range studies on this plant reaches the mankind efficiently. OBJECTIVE(S): The present study was focussed on antihyperglycemic potential of A. reticulata leaves under in vitro and in vivo. MATERIAL AND METHODS: The in vitro phytochemical analysis, total phenolic, flavonoid content, inhibition activity on alpha amylase and alpha glucosidase enzymes were determined for various solvent extracts, followed by in vivo oral toxicity, short term, dose dependant antihyperglycemic studies, oral glucose tolerance tests were performed. The activity of methanolic extract of A.reticulata (MeEAR)-500 mg/kg b.wt was studied for 28 days in diabetic rat model. Histopathological examinations and serum biochemical assays were performed. Gas chromatography-Mass spectrometry (GC-MS) analysis was performed to identify the compounds present in MeEAR. RESULTS: Among the various extracts, MeEAR possesses higher amount of phenols and flavonoids with effective inhibition on carbohydrate hydrolysing enzymes (P < 0.05) and also exhibited higher glycemic control in vivo, with simultaneous improvement in the hepatic and renal activities in diabetic rats. GC-MS analysis revealed the presence of 63 bioactive compounds including carboxylic-acids, alcoholic groups, fattyacid esters, amino acid derivatives. CONCLUSION: Altogether, our study demonstrated that leaves of A.reticulata possess better antihyperglycemic activity and could be developed in to a potential antidiabetic drug with further studies.

Evaluation of wound healing activity of ethanol extract of Annona reticulata L. leaf both in vitro and in diabetic mice model.

BACKGROUND AND AIM: The leaves of AnnonareticulataLinn (niú x inguǒ; Bullock's heart), a member of Annonaceae family, have been used extensively in folk medicine; however, its wound healing potential is yet to be explored. Our aim was to investigate the wound healing ability of A. reticulataleaf extract in vitro and in streptozotocin induced diabetic mice model. MATERIAL AND METHODS: We observed the plant extract induced proliferation and migration of primary human dermal fibroblast (HDF), human skin fibroblast cell line (GM00637) and human keratinocyte cell line (HACAT). The expression of transforming growth factor beta (TGF-ß), connective tissue growth factor (CTGF), vascular endothelial growth factor (VEGF), alpha smooth muscle actin (α-SMA), matrix metalloproteinases (MMP-2, MMP-9), collagen-1, collagen-3, focal adhesion kinase (FAK) were evaluated by Western blot and gelatin zymography. Excisional diabetic wound model was used for in vivo wound healing assay. Furthermore, we processed wound tissue for histological and immunohistochemical study. RESULT: A. reticulata L. leaf extract stimulates proliferation and migration of HDF, skin fibroblast and keratinocyte significantly in a dose dependent manner; expression of TGF-ß, CTGF, VEGF, α-SMA, MMP-2, MMP-9, collagen-1, collagen-3, FAK increased. Additionally, an enhanced expression of phospho-SMAD2, phospho-SMAD3 in the treated cells indicated the activation of TGF-ß signal transduction pathway, similarly increased expression of phospho-AkT suggested activation of PI3/AkT pathway. Expression of CTGF and α-SMA was also increased significantly in wound tissue. Mass spectrometric analysis revealed that mainly two compounds to be present in the extract: quercetin and ß-sitosterol. CONCLUSION: Collective data suggest that A.reticulata leaf extract may have a stimulatory effect in diabetic wound healing.

Complete chloroplast genome sequence and phylogenetic analysis of Annona reticulata.

Annona reticulata is native to South and Central America which has many phytochemical and pharmacological activities suggesting a wide range of clinical application in lieu of cancer chemotherapy. This study provides abundant genomic data for the genetic relationship study, germplasm resources evaluation and varieties selection of A. reticulata. The complete chloroplast genome of A. reticulata was sequenced, assembled, and annotated in this study. The genome size was 201,906 bp and was divided into four regions: a large single-copy region of 69,650 bp, a small single-copy region of 3,014 bp, and two inverted repeat regions of 64,621 bp. A total number of 164 genes were annotated, including 115 protein-coding genes, one pseudogene, 40 tRNA genes, and eight rRNA genes. In terms of gene function, the 164 genes were divided into four major groups: genes for self-replication, photosynthesis, unknown function, and other genes. A maximum likelihood tree based on the chloroplast genome sequences of 24 plant species was constructed. The result of phylogenetic analysis showed that A. cherimola had the closest relationship with A. reticulate.

Antidiabetic, Antihyperlipidemic, Antioxidant, Anti-inflammatory Activities of Ethanolic Seed Extract of Annona reticulata L. in Streptozotocin Induced Diabetic Rats.

Annona reticulata L. (Bullock's heart) is a pantropic tree commonly known as custard apple, which is used therapeutically for a variety of maladies. The present research was carried out to evaluate the possible protective effects of Annona reticulata L. (A. reticulata) ethanolic seed extract on an experimentally induced type 2 diabetes rat model. Male Albino Wistar rats were randomly divided into five groups with six animals in each group viz., control rats in group I, diabetic rats in group II, diabetic rats with 50 and 100 mg/kg/bw of ethanolic seed extract of A. reticulata in groups III and IV, respectively, and diabetic rats with metformin in group V. Treatment was given for 42 consecutive days through oral route by oro-gastric gavage. Administration of A. reticulata seed extract to diabetes rats significantly restored the alterations in the levels of body weight, food and water intake, fasting blood glucose (FBG), insulin levels, insulin sensitivity, HbA1c, HOMA-IR, islet area and insulin positive cells. Furthermore, A. reticulata significantly decreased the levels of triglycerides, cholesterol, LDL, and significantly increased the HDL in diabetic rats. A. reticulata effectively ameliorated the enzymatic (ALT, AST, ALP, GGT) and modification of histopathological changes in diabetic rats. The serum levels of the BUN, creatinine levels, uric acid, urine volume, and urinary protein were significantly declined with a significant elevation in CCr in diabetic rats treated with A. reticulata. MDA and NO levels were significantly reduced with an enhancement in SOD, CAT, and GPx antioxidant enzyme activities in the kidney, liver, and pancreas of diabetic rats treated with A. reticulata. Diabetic rats treated with A. reticulata have shown up-regulation in mRNA expression levels of nuclear factor erythroid 2-related factor 2 (Nrf2), NAD(P)H:quinone oxidoreductase 1 (NQO1), Heme oxygenase-1 (HO-1) and protein expression level of Nrf2 with diminution in Keap1 mRNA expression level in pancreas, kidney, and liver. From the outcome of the current results, it can be inferred that seed extract of A. reticulata exhibits a protective effect in diabetic rats through its anti-diabetic, anti-hyperlipidemic, antioxidant and anti-inflammatory effects and could be considered as a promising treatment therapy in the treatment of diabetes mellitus.

Phytochemical Profile, Toxicity, and Pharmacological Potential of Peels from Four Species of Tropical Fruits.

Tropical fruit peels are generally discarded as waste, yet they contain bioactive substances that could have various uses; in addition, their pharmacological potential remains unexplored. This study aims to characterize the phytochemical profile, toxicity, and pharmacological potential of methanol extracts obtained from the peels of the following tropical fruit species: Annona squamosa L. (purple sugar apple), Annona reticulata L. (custard apple), Chrysophyllum cainito L. (green star apple), and Melicoccus bijugatus Jacq. (mamoncillo). Methanol peel extracts were obtained by maceration. All extracts contained flavonoids, anthraquinones, and triterpenoids as determined by colorimetric methods. A. squamosa and C. cainito exhibited the highest content of total phenols as assayed by the Folin-Ciocalteu method. M. bijugatus showed the highest content of total sugars (fructose, glucose, and sucrose) as determined by high-performance liquid chromatography. A. squamosa and C. cainito presented the highest antioxidant capacities (according to 2,2'-diphenyl-1-picrylhydrazyl, 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid, and cupric reducing antioxidant capacity assays), displayed moderate toxicity against HCT-116 cells, and increased the vinblastine susceptibility of MCF-7/Vin+. A. squamosa and M. bijugatus extracts demonstrated modulation of acetylcholinesterase activity, whereas those of A. reticulata showed anti-inflammatory activity by inhibiting protein denaturation. These results confirm that tropical fruit peels can be valuable sources of bioactive compounds, and our findings provide new information about their pharmacologic potential so that they can be used as raw material for the development of new drugs aimed at treating a variety of ailments.

Glycogen-gold nanohybrid escalates the potency of silymarin.

In this study, a glycogen-gold nanohybrid was fabricated to enhance the potency of a promising hepatoprotective agent silymarin (Sly) by improving its solubility and gut permeation. By utilizing a facile green chemistry approach, biogenic gold nanoparticles were synthesized from Annona reticulata leaf phytoconstituents in combination with Sly (SGNPs). Further, the SGNPs were aggregated in glycogen biopolymer to yield the therapeutic nanohybrids (GSGNPs). Transmission electron microscopy, UV-Vis spectroscopy, X-ray diffraction, and Fourier transform infrared spectroscopy analysis confirmed the successful formation and conjugation of both SGNPs and GSGNPs. The fabricated nanohybrids showed significant protection against CCl4-induced hepatic injury in Wistar rats and maintained natural antioxidant (superoxide dismutase and catalase) levels. Animals treated with GSGNPs (10 mg/kg) and SGNPs (20 mg/kg) retained usual hepatic functions with routine levels of hepatobiliary enzymes (aspartate transferase, alanine transaminase, alkaline phosphatase, and lactate dehydrogenase) and inflammatory markers (interleukin-1ß and tumor necrosis factor-α) with minimal lipid peroxidation, whereas those treated with 100 mg/kg of Sly showed the similar effect. These results were also supported by histopathology of the livers where pronounced hepatoprotection with normal hepatic physiology and negligible inflammatory infiltrate were observed. Significant higher plasma Cmax supported the enhanced bioavailability of Sly upon GSGNPs treatment compared to SGNPs and free Sly. Graphite furnace atomic absorption spectrophotometry analysis also substantiated the efficient delivery of GSGNPs over SGNPs. The fabricated therapeutic nanohybrids were also found to be biocompatible toward human erythrocytes and L929 mouse fibroblast cells. Overall, due to increased solubility, bioavailability and profuse gut absorption; GSGNPs demonstrated tenfold enhanced potency compared to free Sly.