Asian Pacific Journal of Tropical Biomedicine

469-480

Authors: Siti Mariam A. Lathiff, Norzafneza M Arriffin, Shajarahtunnur Jamil

This article aims to review the scientific data on phytochemical and pharmacological studies of Artocarpus collected from Malaysia as well as to highlight their usage as ethnomedicine worldwide. About 55 Artocarpus species are distributed worldwide and 32 of the Artocarpus species can be found in Malaysia. Artocarpus species are well known worldwide for their edible fruits such as Artocarpus heterophyllus (jackfruit), Artocarpus integer (chempedak), and Artocarpus communis (breadfruit). Aside from its edible fruits, the timber is valued for light constructions, crates, large canoes, and boats. The literature for this review was searched using the term ‘Artocarpus’, ‘Artocarpus Malaysia’, ‘Artocarpus extracts’, ‘Artocarpus traditional medicine’ and ‘Artocarpus ethnomedicine’ from published books and scientific journals via various engines such as The Web of Science, PubMed, Science Direct, Scopus, Research Gate, and Google Scholar. The references cited from the retrieved articles were also scanned and cross-checked. All published studies on phytochemical and pharmacological activities of Malaysia’s Artocarpus species up to January 2021 were included in this review. Articles on phytochemical studies of Malaysia’s Artocarpus revealed the isolation of flavonoids as the major constituents. Research on pharmacological activities of the isolated phytochemicals showed that these compounds exhibited significant disease-linked-enzyme (tyrosinase, cholinesterase, glucosidase) inhibitors as well as antioxidant, anti-inflammatory, antimicrobial, and cytotoxic activities. The ethnomedicinal data gathered are useful to understand and prioritize Artocarpus species that can contribute to potent phytochemicals and possibly new drug leads. This review also provides valuable information for the future development of isolated compounds from Artocarpus species.

491-499

Authors: Sahar M Jaffal, Sawsan A Oran, Mohammad I Alsalem

Objective: To explore the anti-inflammatory and antipyretic effects of methanolic leaf extract from Arbutus andrachne and its mechanism of action. Methods: Paw edema was induced by intraplantar (i.pl.) injection of λ-carrageenan (1% w/v, 100 μL/paw) while pyrexia was evoked by intraperitoneal (i.p.) injection of 20% baker’s yeast (20 mL/kg body wt) in male Wistar rats. The anti-inflammatory and antipyretic effects of Arbutus andrachne methanolic leaf extract were explored by injecting rats with different doses of the plant extract (150, 300, and 600 mg/kg body wt, i.p.). Selective antagonists for transient receptor potential vanilloid-1 (TRPV1), cannabinoid receptor 1 (CB1), and alpha-2 adrenergic receptor (a2-AR) were used to unravel the extracts’ mechanism of action. Blood samples were collected from the heart of rats to measure the levels of interleukin-6 (IL-6) and prostaglandin E2 (PGE2) by enzyme-linked immunosorbent assay. Results: The extract exhibited potent anti-inflammatory activity by decreasing paw thickness and IL-6 levels. In addition, yeast-evoked pyrexia was attenuated by the extract treatment via TRPV1 and CB1 receptors and a reduction in PGE2 levels. No significant effects were found for a2-AR. Moreover, the rats that received the plant extract demonstrated similar responses to the positive control group. Conclusions: Arbutus andrachne can be a good candidate for treating inflammation and pyrexia and should be further investigated.

510-518

Authors: Hitendra S Mahajan, Nayana D Patil

Objective: To characterize a nanoemulsion containing a synergistic combination of curcumin (CUR) and quercetin (QUE) for brain targeting through the nose. Methods: The synergistic activity of the combination of two phytoconstituents, curcumin and quercetin (CUR-QUE) was determined at a 2:1 ratio. Nanoemulsions containing the synergistic combination were prepared using high-pressure homogenization technique. Nanoemulsion formulation was characterized for globule size, drug content, thermodynamic stability, zeta potential measurement, and drug release. Histological studies were performed using isolated nasal mucosa of sheep. Moreover, in vivo studies using allograft model were performed. Results: CUR-QUE in a 2:1 ratio combination showed the lowest combination index values 0.99 (combination index<1) for treatment of human glioblastoma U373-MG cell. The combination inhibited the growth of human glioblastoma U373MG cells. The significant drug targeting efficiency percentage (178.25±2.86 for CUR and 170.54±6.11 for QUE) and nose to brain drug direct transport percentage (44.05±0.93 for CUR and 38.25±0.83 for QUE) of optimized nanoemulsion (F9) indicated effective central nervous system targeting via the intranasal route. In vivo anticancer study using allograft model of nude mice demonstrated potential anticancer activity of the synergistic combination of CUR-QUE as compared to doxorubicin. Conclusions: This study demonstrates that synergistic combination of CUR-QUE was optimal for treatment of human glioblastoma. Moreover, significant site-specific delivery to the brain was achieved by administering nanoemulsion through the nose. The study also reveals that intranasal delivery of nanoemulsion containing synergistic combination of CUR and QUE could be a promising strategy for brain tumor treatment.

421-428

Authors: Amin F Majdalawieh, Sarah M Yousef, Imad A Abu-Yousef

The ongoing outbreak of novel coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has caused an unprecedented global health crisis. Development of a cure for this devastating disease is currently at full speed, with several vaccines against COVID-19 already authorized and administered. Currently, demand for these vaccines far exceeds supply. As such, boosting immunity represents a viable route to halt the rapid spread of SARS-CoV-2 and limit fatalities until vaccines become more readily and widely available. The use of phytochemicals appears to be a promising panacea. Sesamin, a lignan isolated from Sesamum indicum seeds, is known for its potent pharmacological properties, and is therefore hypothesized as a potential candidate in the therapeutic regimen against COVID-19. Herein, we highlight the confirmed therapeutic anti-inflammatory and immune-modulatory potential of sesamin against myriads of respiratory disorders, and tentatively suggest that sesamin may exert similar potent effects against COVID-19. Precisely, we speculate that sesamin may help alleviate COVID-19 via restoring Th1/Th2 balance and preventing inflammation and cytokine storm development. Additionally, we further support the promising role of sesamin against COVID-19 by underscoring the direct evidence, which suggests that sesamin may demonstrate promising inhibitory potential against three important SARS-CoV-2 targets, namely main protease, spike protein, and angiotensin-converting enzyme 2 receptor. Although preliminary, there is ample evidence to propose sesamin as a potential phytotherapeutic and prophylactic candidate against COVID-19. Further in vitro, in vivo, and preclinical studies are required to further substantiate the role of sesamin in the prevention and/or treatment of COVID-19.

440-449

Authors: Brenda Siringoringo, Nawiya Huipao, Chittipong Tipbunjong, Jongdee Nopparat, Santad Wichienchot, Albert M Hutapea, Pissared Khuituan

Objective: To investigate the effect of Gracilaria fisheri oligosaccharides (GFO) on inflammation and colonic epithelial barrier dysfunction in colitis mice. Methods: The animals were treated by oral gavage with distilled water, 1 000 mg/kg inulin, 100, 500, or 1 000 mg/kg GFO for 14 d, or treated with 50 mg/kg mesalamine for 5 d after colitis induction (on day 10). Histopathology, inflammatory cytokines, colonic permeability, and tight junction proteins were investigated by hematoxylin and eosin staining, immunohistochemical staining, Ussing chamber technique, and Western blotting assays, respectively. Results: GFO ameliorated histological damage in colitis mice when compared to untreated colitis mice. Treatments with 100, 500, and 1 000 mg/kg GFO reduced TNF-α expression, while IL-1β was significantly reduced in colitis mice treated with 500 and 1 000 mg/kg. Compared to untreated colitis mice, GFO increased transepithelial electrical resistance, reduced fluorescein isothiocyanate-dextran paracellular flux, and modulated tight junction proteins (occludin and claudin 2) in colitis mice. Conclusions: GFO has anti-inflammatory activity and could modulate colonic epithelial barrier dysfunction in acetic acid-induced colitis mice. Furthermore, GFO could modulate the expression of tight junction proteins that play important roles in colonic barrier function.

460-468

Authors: Maria Carmen S. Tan, Mary Stephanie S. Carranza, Virgilio C Linis, Raymond S Malabed, Yves Ira A. Reyes, Francisco C Franco, Glenn G Oyong

Objective: To evaluate antioxidant, cytotoxic, and anti-venom capacity of crude bark extracts of Alstonia parvifolia Merr. Methods: Gas chromatography-mass spectrometry (GC-MS) and energy dispersive X-ray analyses were accomplished to characterize the chemical constituents of Alstonia parvifolia. Biochemical characterization was evaluated using an inhibitory phospholipase A2 (PLA2) assay, DPPH, and cytotoxicity assays. Using the constituents listed in the GC-MS analyses, molecular docking was conducted to inspect the binding energies between the chosen compounds and selected PLA2 isoforms. Results: GC-MS analyses showed that the Alstonia parvifolia crude extract consisted predominantly of acetylmarinobufogenin (14.89%), γ-sitosterol (10.44%), 3-O-methyl-D-glucose (5.88%), 3,5-dimethoxy-4-hydroxyphenylacetic acid (5.30%), (2α,5α)-17-methoxyaspidofractinin-3-one (AFM) (4.08%), and 2,3,5,6,7,8,9-heptahydro-1-phenyl-5-(p-chlorophenylimino)-1H-benzo[e][1],[4]thiazepine (HPT) (1.37%). The principal elemental components of Alstonia parvifolia were Ca (4.012%) and K (1.496%), as exhibited by energy dispersive X-ray examination. Alstonia parvifolia showed significant free radical scavenging ability (IC50: 0.287 mg/mL) and was non-cytotoxic to normal HDFn cells (IC50 >100 μg/mL). Moreover, Alstonia parvifolia was favorably cytotoxic to MCF-7 (IC50: 4.42 μg/mL), followed by H69PR, HT-29, and THP-1, with IC50 values of 4.94, 5.07, and 6.27 μg/mL, respectively. Alstonia parvifolia also displayed notable inhibition against PLA2 activity of Naja philippinensis Taylor venom with IC50 of (15.2 ± 1.8) μg/mL. Docking and cluster analyses projected negative binding energies from AFM (-6.36 to -9.68 kcal/mol), HPT (-7.38 to -9.77 kcal/ mol), and acetylmarinobufogenin (-7.22 to -9.59 kcal/mol). These calculations were for the particular interactions of Alstonia parvifolia constituents to PLA2 homologues where the utmost affinity was detected in HPT owing to the dipole interactions with amino acid residues. Conclusions: The bark extract of Alstonia parvifolia shows great potential as an anti-venom agent due to its low cytotoxic profile, remarkable PLA2 inhibition, and docking binding energies between its bioactive constituents and PLA2 homologues.

1-9

Authors: Lucia Dwi Antika, Rita Marleta Dewi

Over the past decades, epidemiological studies have concluded that a diet rich in plant-derived products plays a pivotal role in human health. Fisetin (3,3’,4’,7-tetrahydroxyflavone) is a hydrophobic polyphenolic compound primarily found in edible plants (e.g. strawberry, blueberry, apple, grape, persimmon, kiwi, and cucumber). Various preclinical studies have revealed that fisetin exhibits a wide range of pharmacological effects such as antioxidant, anti-inflammatory, anti-carcinogenic, anti-osteoporotic, antimicrobial, and anti-diabetic properties. Therefore, the pharmacological in vitro and in vivo studies on fisetin are discussed in this review. Additionally, this review would be useful for further study regarding the potential of natural products, notably fisetin, and its therapeutic potential for the prevention and treatment of diseases.

20-28

Authors: Vikas Kushwaha, Sukhbir Kaur

Objective: To evaluate the immunostimulatory potential of cross-reactive molecule heat shock protein 60 (HSP60) of filarial parasite Brugia malayi and Leishmania donovani. Methods: HSP60 of Brugia malayi (BmHSP60) was amplified using gene-specific primer, cloned in pTriEx4 vector, expressed in BL21-DE3 cells, and recombinant HSP60 (rHSP60) of ~65 kDa was purified by affinity chromatography using Ni-NTA column. The recombinant protein was desalted by the dialysis membrane, and the presence of endotoxin level was determined by Limulus amebocyte lysate assay. The recombinant protein was tested for cell proliferation, nitric oxide release, expression of Th1 and Th2 cytokines, and transcription factors (STATs) in vitro using murine macrophage cell line (J774A.1). Results: Higher cell proliferation indicated that BmHSP60 had immunostimulatory potential. rBmHSP60 exposure upregulated the expression of iNOS, STAT1, STAT4, Th1 cytokines (IFN–γ, TNF–α, IL-12), and nitric oxide release. In addition, no remarkable change was observed in the expression of IL-6, IL-10, and STAT3 in macrophage cell line J774A.1. The ELISA analysis showed the levels of IFN-γ, TNF-α, and IL-12 were upregulated while IL-10 level was downregulated, revealing that BmHSP60 triggered a Th1 immune response. Conclusions: Our study demonstrates that rBmHSP60 has immunogenic properties which effectively enhances the Th1 type immune responses, and can be used as an immunoprophylactic agent against leishmaniasis. Furthermore, in vivo studies are in progress to determine the protective role of rBmHSP60 against Leishmania donovani infection in a mouse model.

40-46

Authors: Mohammad Javanbakht, Abedin Saghafipour, Keyvan Ezimand, Amir Hamta, Leyli Zanjirani Farahani, Nazanin Soltani

Objective: To determine the effect of climatic and environmental factors on the incidence of cutaneous leishmaniasis in Qom province in 2018. Methods: In this cross-sectional study, the data on cutaneous leishmaniasis incidence were collected from the Disease Control and Prevention Center in Qom province. Climatic and environmental data including Normalized Difference Vegetation Index (NDVI), Land Surface Temperature (LST), and soil moisture were extracted using satellite images. Data of altitude and sunny hours were provided based on shuttle radar topography mission digital elevation model and hemispherical viewshed algorithm, respectively. The associations of climatic and environmental variables with the incidence of the disease were analyzed by Pearson correlation method. The ArcGIS 10.3 software was used to determine the geographical distribution of these factors. Results: There were positive correlations between cutaneous leishmaniasis incidence and the two climatic factors: LST and sunny hours per day (P=0.041, P=0.016), and it had weak negative correlations with the digital elevation model (P=0.27), soil moisture (P=0.54), and NDVI (P=0.62). The time delay analysis showed that in one-, two-, and three month periods, the correlations increased with a 95% confidence interval. Accordingly, the correlation with the three-month time delay was positive and relatively strong between the cutaneous leishmaniasis incidence and LST and sunny hours (P=0.027, P=0.02); nevertheless, there were negative correlations between the cutaneous leishmaniasis incidence and the soil moisture (P=0.27) and NDVI (P=0.62). Conclusions: As Qom is located in one of the semi-arid climate zones, topography and solar energy are important factors affecting the incidence of cutaneous leishmaniasis in autumn. Therefore, appropriate disease control programs are recommended.

387-396

Authors: Benjamin Kingsley Harley, Rita Akosua Dickson, Isaac Kingsley Amponsah, Robert A Ngala, Dorice Berkoh, Theophilus Christian Fleischer

Objective: To investigate the in vivo and in vitro antidiabetic potential of Chrysophyllum albidum. Methods: The effects of oral treatment with hydro-ethanolic extract (125, 250 and 500 mg/kg) of the stem bark of Chrysophyllum albidum and glibenclamide for 21 d on glucose level, serum enzyme markers for liver function, lipid profile, total protein, serum urea, serum creatinine, and body weight were evaluated in experimental diabetic rats administered with 45 mg/kg of streptozotocin. In vitro assays including glucose uptake in C2C12 cells and 3T3-L1 adipose tissues, α-glucosidase and α-amylase inhibition were employed to evaluate the possible mechanism of hypoglycemic action of the extract. DPPH and nitric oxide radical antioxidant activity of the extract was also measured. Results: The increased levels of blood glucose, triglycerides, low- density lipoprotein, total cholesterol, serum aspartate, and alanine transaminases, creatinine, and urea in the diabetic animals were reduced significantly (P<0.01) after treatment with Chrysophyllum albidum extract. The decreased total protein and high-density lipoprotein concentrations were normalized after treatment. In addition, the extract significantly (P<0.01) increased the transport of glucose in 3T3-L1 cells and C2C12 myotubes and exhibited considerable potential to inhibit α-amylase and α-glucosidase. It also demonstrated potent antioxidant action by scavenging considerably DPPH and nitric oxide radicals. Conclusions: Chrysophyllum albidum stem bark extract exhibits considerable antidiabetic effect by stimulating glucose uptake and utilization in C2C12 myotubes and 3T3-L1 adipocytes as well as inhibiting the activities of α-amylase and α-glucosidase.

403-410

Authors: Chi-Yeol Yoo, Hyeong-U Son, Alshammari Fanar, Hee-Jung Choi, Md Badrul Alam, Sang-Han Lee

Objective: To investigate the effect of Opuntia humifusa aqueous extract on gastric ulcers. Methods: An ethanol-induced model was used to examine the protective effect of Opuntia humifusa against gastric ulcers. The gastric ulcer index was evaluated via clinical observation and image analysis. Various inflammatory indicators were determined by RT-PCR and Western blotting assays. Results: The gastric ulcer index was reduced to 8% in the group treated with Opuntia humifusa aqueous extract compared with that in the control group. RT-PCR analysis revealed that MUC5AC expression was reduced to 39% in the control group compared with the non-treated group, whereas the omeprazole and Opuntia humifusa aqueous extract-treated groups increased the expression to 95% and 79%, respectively. Moreover, the expressions of various cytokines including TNF-α, IL-1β, and IL-6 were increased in the control group, while decreasing in Opuntia humifusa aqueous extract-treated group. Opuntia humifusa aqueous extract also suppressed the expressions of iNOS, COX-2, and its transcription factor NF-κB and increased mucus content considerably as compared to the control group. Conclusions: These results suggest that Opuntia humifusa aqueous extract is suitable as an alternative remedy for gastric ulcer treatment.

417-425

Authors: Riffat Batool, Ejaz Aziz, Javed Iqbal, Hina Salahuddin, Benny Kwong-Huat Tan, Saira Tabassum, Tariq Mahmood

Objective: To explore antioxidant potential, anti-cancer activity, and phytochemicals of Commelina benghalensis L. Methods: The roots of Commelina benghalensis were extracted in different solvents (methanol, ethanol, benzene, chloroform, n-hexane) with a range of polarity. Antioxidant activity was evaluated by reducing power assay, DPPH radical scavenging activity and phosphomolybdenum method, cytotoxicity by MTT assay, apoptotic and cell cycle analysis by flow cytometry, migratory and invasive potential by wound scratch assay and invasion assay, respectively, functional groups analysis by FT-IR spectroscopy and phytochemicals by aluminum chloride colorimetric and Folin- Ciocalteu methods. Results: The extracts showed worthy antioxidant potential. The chloroform extract demonstrated the most significant cytotoxic effect on MDA-MB-231 (breast cancer) cell line, induced apoptosis and reduced migratory and invasive potential of MDA-MB-231 cells. Methanol and ethanol extracts presented good yield of total phenolic and total flavonoid contents. The FTIR spectroscopic studies revealed different characteristic peak values with various functional compounds such as alkenes, alkanes, aliphatic amines, aromatics, alkyl halides, carboxylic acid, alcohols, ester, aldehydes and ketones. Conclusions: The results demonstrate the potential use of Commelina benghalensis as a good antioxidant with significant anti-cancer effect.

341-342

Authors: Upasana Shyamsunder Singh, Aparup Das

None

353-360

Authors: Syed Ali Raza, Ayoub Rashid Chaudhary, Muhammad Waseem Mumtaz, Ahmad Adnan, Hamid Mukhtar, Muhammad Tayyab Akhtar

Objective: To profile the secondary metabolites and to evaluate the antidiabetic potential of hydroethanolic leaf extracts of Conocarpus lancifolius. Methods: The various hydroethanolic extracts of Conocarpus lancifolius leaf were prepared by ultrasonication assisted freeze- drying. Total phenolic contents, flavonoid contents, antioxidant activity, α-glucosidase and α-amylase inhibitions of leaf extracts were determined. The metabolite profiling was accomplished by UHPLC-Q-TOF-MS/MS analysis. The antidiabetic assessment of the most potent extract was carried out by measuring the hypoglycemic and hypolipidemic effect in the high fat diet-fed diabetic albino mice. The blood glucose level, haemoglobin, total cholesterol, high-density lipoproteins (HDL) and low-density lipoproteins (LDL) were determined. Results: The 60% ethanolic extract exhibited the highest phenolic and flavonoid contents of (349.39 ± 2.13) mg GAE/g dry extract and (116.95 ± 2.34) mg RE/g dry extracts, respectively, and the highest DPPH scavenging activity with an IC50 value of (32.87 ± 1.11) μg/ mL. The IC50 values for α-glucosidase and α-amylase inhibitions were (38.64 ± 0.93) μg/mL and (44.80 ± 1.57) μg/mL, respectively. UHPLC-Q-TOF-MS/MS analysis confirmed the presence of gallic acid, ellagic acid, corilagin, kaempherol-3-O-rutinoside, caffeic acid derivative, isorhamnetin and galloyl derivatives in the 60% ethanolic extract. Plant extract at a dose of 450 mg/kg body weight reduced blood glucose level, total cholesterol, LDL and HDL, and increased haemoglobin in alloxan-induced diabetic mice, Conclusions: Conocarpus lancifolius leaves are proved as a good source of biologically functional metabolites and possess antidiabetic activity which may be further explored to treat diabetes.

369-378

Authors: Somprasong Saenglee, Gulsiri Senawong, Jarckrit Jeeunngoi, Sanun Jogloy, Albert J Ketterman, Banchob Sripa, Thanaset Senawong

Objective: To investigate the effect of combination treatments of cisplatin and KK4 and ICG15042 peanut testa extracts against cholangiocarcinoma cells in vitro. Methods: The growth inhibition, cell cycle arrest and apoptosis of cholangiocarcinoma cells were determined by 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and flow cytometry analysis, respectively. The levels of proteins involved in apoptosis were assessed using Western blotting assays. The caspase activity was assessed using a colorimetric caspase activity assay. Results: Cisplatin and peanut (KK4 and ICG15042) testa extracts inhibited the growth of cholangiocarcinoma cell lines (KKU- M214 and KKU-100 cells) in a dose- and time-dependent manner. The combination treatments reduced cell viability and induced apoptosis of cholangiocarcinoma cells more efficiently than singledrug treatments. Cancer cell death synergistically mediated by cisplatin and peanut testa extracts was observed in KKU-M214 cells (combination index < 1.0) but not in KKU-100 cells (combination index > 1.0). The combination treatments also increased the sub- G1 population and caused KKU-M214 cell cycle arrest at S and G2/ M phases, which were the combined effects of cisplatin (S phase arrest) and peanut testa extracts (G2/M phase arrest). In addition, pERK1/2, Ac-H3, Bcl-2 and proteins related to apoptosis, including Bax and caspases 3, 8, 9, exhibited enhanced expression in KKU- M214 cells. The combination treatments caused down-regulation of p53, whereas the expression of p21 was fairly constant when compared with cisplatin single drug treatment. Conclusions: Peanut testa extracts in combination with cisplatin synergistically reduce cell viability and induce apoptosis through stimulation of caspases 3, 8 and 9 in KKU-M214 cells.