Asian Pacific Journal of Tropical Biomedicine

188-195

Authors: Ramakrishnan Sabitha, Kumari Nishi, Vinoth Prasanna Gunasekaran, Govindhan Annamalai, Balupillai Agilan, Mathan Ganeshan

Objective: To examine the effects of p-coumaric acid on ethanol-induced kidney injury in Swiss Wistar rats. Methods: Ethanol (25% v/v) was used to induce nephrotoxicity in rats. p-Coumaric acid was orally administered at 50, 100, or 200 mg/kg body weight. The levels of oxidative parameters were determined; pro-inflammatory biomarkers were analyzed by Western blotting and apoptotic protein was analyzed by immunohistochemistry. Results: Ethanol treated rats showed decreased levels of antioxidants and aberrant production of pro-inflammatory cytokines (IL-6, IL1β, TNF-α), NF-κB activation and imbalance of proand anti-apoptotic proteins (Bcl-2, Bax, caspase 3). Meanwhile, p-coumaric acid restored antioxidant levels and decreased the levels of inflammatory cytokines, NF-κB, and proapoptotic proteins and increased Bcl-2 expression. Conclusions: p-Coumaric acid ameliorates ethanol-induced kidney injury by restoring antioxidant production and suppressing cellular apoptosis and inhibiting NF-κB expression. p-Coumaric acid should be further investigated as a promising candidate for ethanol-induced kidney toxicity.

204-208

Authors: Ratree Tavichakorntrakool, Aroonlug Lulitanond, Arunnee Sangka, Seksit Sungkeeree, Natthida Weerapreeyakul

Objective: To evaluate antibacterial activity and the bioactive compounds of 50% hydroethanolic extract of Alpinia zerumbet (A. zerumbet) rhizomes. Methods: Eight reference microbial strains including two Gram-positive bacteria [Staphylococcus aureus (ATCC 29213) and Enterococcus faecalis (ATCC 29212)] and six Gram-negative bacteria [Escherichia coli (ATCC 25922), Klebsiella pneumoniae (ATTC 700603), Proteus mirabilis (DMST 8212), Salmonella enterica subsp. enterica serovar Vellore. (ATCC 15611), Shigella flexneri (ATCC 12022) and Pseudomonas aeruginosa (ATCC 27853)], were used to test antimicrobial susceptibility by the broth microdilution method. Bioactive compounds were analyzed by using HPLC. Results: The minimum inhibitory concentration values of A. zerumbet extract were 8 mg/mL for Staphylococcus aureus, Escherichia coli and Shigella flexneri and 16 mg/mL for Enterococcus faecalis and the other four Gram-negative bacilli. HPLC chromatograms revealed that the A. zerumbet extract contained hydroxybenzoic acids, hydroxycinnamic acids and flavonoids. Conclusions: The constituents of A. zerumbet rhizomes could be a potential source of antibacterial compounds, warranting further study of A. zerumbet extract.

217-221

Authors: Umay Chen, Md. Saddam Hussain, Tanoy Mazumder, SM Naim Uddin, Sujan Banik

Objective: To evaluate the neuropharmacological properties of Costus speciosus (C. speciosus) rhizome using different experimental mouse models. Methods: The anxiolytic effect was investigated by hole-board test, elevated plus maze and light/dark test, while central nervous system (CNS) depressant effect was evaluated by thiopental sodium-induced sleep test. Finally, antidepressant effect was evaluated by forced swimming test and tail suspension test. Results: In both elevated plus maze and hole board test, 400 mg/kg C. speciosus showed more significant CNS depressant effect than 1 mg/kg diazepam. Both 200 mg/kg and 400 mg/kg C. speciosus extract produced a significant dose-dependent decrease in onset of sleep. In forced swimming test, C. speciosus rhizome showed a decrease in duration of immobility in a dose-dependent manner. Imipramine (10 mg/kg) and C. speciosus extract at 400 mg/kg dose exhibited a significant reduction in duration of immobility in tail suspension test which provided additional evidence of antidepressant effect of C. speciosus rhizome. Conclusions: Our study indicates that C. speciosus rhizome possesses CNS depressant, anxiolytic and antidepressant-like activities. Further studies are warranted determine the exact phytoconstituents and mechanism of action responsible for the neuropharmacological effect.

140-149

Authors: Nurul Hazirah Che Hamzah, Arifullah Mohammed, KNS Sirajudeen, Mohd Asnizam Asari, Zulhazman Hamzah, Ibrahim Khalivulla Shaik

Objective: To investigate the wound-healing effect of Alocasia longiloba (A. longiloba) petiole extract on wounds in rats. Methods: Twenty-two male Sprague-dawley rats were randomly assigned to receive 10% solcoseryl gel, phosphate buffer saline, 50% ethanol, 95% ethanol and hexane extracts of A. longiloba at 1.5%, 3% and 6% doses, respectively. A full thicknesses wound (6 mm) was created on the dorsal of the rat; and all rats were applied with the extract solutions, 10% solcoseryl gel and phosphate buffer saline once a day topically until day 12. The wound was photographed on day 1, 6 and 12, and the percentage of wound contraction was calculated. On day 12, rats were sacrificed and histological examination of granulation tissue was carried out using haematoxylin & eosin and Masson’s Trichrome stain to determine the wound healing effect. Results: In this study, 6% of 50% and 95% ethanol extracts of A. longiloba showed 82.50% and 82.32% wound contraction, respectively, and were comparable with 10% solcoseryl gel (82.30%). Meanwhile, phosphate buffer saline treated group showed the lowest wound contraction (69.86%). Histological assessment of wound treated with 6% of 95% ethanol extract of A. longiloba showed distinct epidermal and dermal layer, higher proliferation of fibroblast and more angiogenesis with collagen compared to other wound treated groups. Conclusions: A. longiloba petiole extracts have a wound healing potential and 6% of 95% ethanol extract of A. longiloba is more effective. Further studies are required to understand the wound healing mechanism of action of the extract.

158-167

Authors: Ali Abdulqader, Faisal Ali, Amin Ismail, Norhaizan Mohd Esa

Objective: To identify and determine the composition of antioxidant compounds, and to evaluate the antioxidant abilities of Gac fruit parts (peel, pulp, seed and aril) grown in Malaysia. Methods: LC-MS/MS was used for identification of antioxidant compounds and UV-Vis for estimation of the contents of phenolics, flavonoids, and carotenoids. Lycopene and β -carotene were quantified using high-performance liquid chromatography. DPPH (2, 2-diphenyl-1- picrylhydrazyl) and ferric reducing antioxidant power assays were employed to evaluate antioxidant capacities. Results: Phytochemicals were found amongst all the fruit parts. Notably, significant amounts of carotenoids [(107.4 ± 4.5), (85.7 ± 4.4), (110.6 ± 2.1) mg/100 g dry weight (DW)], and relatively high levels of both phenolics [(27.3 ± 1.7), (28.9 ± 2.4), (30.8 ± 2.7) mg/100 g DW] and flavonoids [(38.1 ± 2.2), (8.8 ± 1.3), (24.5 ± 3.3) mg/100 g DW] were found in the fruit’s peel, pulp and aril, respectively. Seed part also showed a relatively high level of flavonoids [(18.1 ± 2.3) mg/100 g DW]. Lycopene and β -carotene were found to be significantly high (P < 0.05) in aril [(579.3 ± 22.7) and (621.0 ± 35.0) μg/g DW], followed by peel [(51.0 ± 7.5) and (210.0 ± 12.5) μg/g DW] and pulp [(37.6 ± 10.9) and (205.6 ± 22.1) μg/g DW)]. Antioxidant assays revealed that aril possessed the highest scavenging activity (IC50 = 865 μg/mL), while the peel possessed the highest ferric reducing power of 140 pmol FeSO4/μg. Conclusions: The current results demonstrate that Gac fruit grown in Malaysia is a rich source of phytochemicals, especially carotenoids, and possesses antioxidant activities. Thus, such findings suggest Gac fruit as a source of an antioxidant plant.

176-180

Authors: Enegide Chinedu, Peter A Akah, Dabum L Jacob, Ifeoma A Onah, Chimere Y Ukegbu, Chukwuma K Chukwuemeka

Objective: To evaluate the antimalarial activity of the ethylacetate and butanol fractions of Combretum nigricans (C. nigricans) leaf extract in mice. Methods: C. nigricans solvent (butanol and ethylacetate) fractions were screened for their phytochemical constituents using standard procedures illustrated by Harborne and Evans. The Peters’ 4-day suppressive test against early malaria infection, Rane’s curative test against established malaria and prophylactic test for residual activity were employed for evaluating the antimalarial potential in mice. Results: The phytochemical screening revealed the presence of alkaloids, terpenoids, saponins, and flavonoids in both fractions at different intensity. Both fractions exhibited significant antimalarial activity in all test models (P<0.05). The ethylacetate fraction of C. nigricans had better chemosuppressive and curative effects compared to the butanol fraction, which however, elicited a better chemoprophylactic effect. The chemosuppressive effect of C. nigricans ethylacetate fraction (200-800 mg/kg) was 77.6%, 69.1% and 86.1%; curative effect was 62.3%, 71.3% and 72.4%; while the chemoprophylactic activity was 32.1%, 48.6% and 61.2% respectively. C. nigricans butanol fraction (200-800 mg/kg) had 40.3%, 54.1% and 69.1% chemosuppression; 26.2%, 36.9% and 34.5% curative effect; and 48.4%, 70.0% and 87.4% chemoprophylaxis. Conclusions: Both solvent fractions of C. nigricans possess antimalarial activity, and may be useful at different stages of malaria therapy.

98-108

Authors: Attia H Atta, Samar M Mouneir, Soad M Nasr, Doaa Sedky, Amany M Mohamed, Shimaa A Atta, Hassan Mohamed Desouky

Objective: To analyze the phytochemical constituents, and to explore potential protective effect of the methanol extract of Moringa oleifera (M. oleifera) seeds and Egyptian propolis, each alone or concurrently administered on acetic acid-induced ulcerative colitis in rats. Methods: Eight groups of 5 rats each were used: normal control group with distilled water, model group, two groups with M. oleifera seeds (100 and 200 mg/kg), two groups with propolis (50 and 100 mg/kg), one group with concurrent administration of both, and one group with prednisolone (reference drug). Macro-and microscopic picture, ulcer index and lesion scores, oxidative markers, inflammatory mediators, in vitro activity of the inflammatory enzymes and 1, 1-diphenyl-2-picrylhydrazyl free radicals scavenging activity were evaluated. The phytochemical constituents of both extracts were explored by GC-MS analysis. Results: Both treatments modulated the macro-and microscopic picture, decreased the ulcerative index, lesion score, oxidative markers and inflammatory mediators, and inhibited the COX-1 and COX-2 enzymes. Propolis appeared to be powerful free radicals scavenger. A powerful synergistic effect of both treatments in modulating the course of the disease was reported. GC-MS analysis of methanol extract of M. oleifera seeds and propolis revealed the presence of 50 and 34 compounds, respectively. Conclusions: M. oleifera seeds and propolis methanol extracts have modulated the course of acetic acid-induced ulcerative colitis. Moreover, both treatments induce a good synergistic effect against the disease. Isolation of the active constituents is recommended.

116-122

Authors: Emin Sengul, Sevda Urçar Gelen, Serkan Yıldırım, Fikret Çelebi, Ali Çınar

Objective: To investigate the effects of probiotic bacteria on cisplatin (CP)-induced nephrotoxicity. Methods: In the present study, 50 Sprague-Dawley rats were used and randomly divided into five groups including control, CP, probiotic bacteria treatment groups with different doses (0.5 and 1 mL) and only probiotic bacteria group. After CP and probiotic administration on seven days, rats sacrificed under anesthesia on the eighth day. The serum urea, creatinine, and blood urea nitrogen levels were analyzed. In renal tissue, malondialdehyde levels, superoxide dismutase and glutathione activity, interleukin-8, interleukin-1β and tumor necrosis factor-alpha levels were determined and histopathological and immunohistochemical changes were also examined. Results: According to results, urea, creatinine and blood urea nitrogen levels as well as kidney weights increased in CP group. Also, CP induced inflammation, oxidative stress, DNA damage and apoptosis in kidney tissue and caused histopathological changes. Administration of the high dose of probiotic bacteria could prevent these changes and damages. Conclusions: This study reveals that probiotic bacteria has protective effects on CP-induced renal damage in rats.

129-134

Authors: Hakan Aydin, Mehmet Ozkan Timurkan, Gulizar Acar Kirmizi

Objective: To investigate the presence, prevalence and phylogenetic classification of bovine torovirus (BToV). Methods: Stool samples from 72 calves, which were negative for primary gastroenteritis agents, were examined with the nested PCR method by using BToV M gene-specific primers. Results: BToV was detected in 12 (16.7%) out of 72 samples. Phylogenetic analysis was performed using nucleotide and amino acid sequences. In the phylogenetic tree, European, American, Far East and Turkish strains were found to be divided into different branches. Interestingly, it was observed that Turkish strains were divided into two subgroups. Considering the amino acid sequences of these strains having differences at nucleotide level, the change at the 3rd amino acid of the partial M gene in Turkish strains has made Turkish strains different from all other strains. Similarly, the differences were observed in the 18th, 20th, 63rd and 93rd amino acids of the partial M gene only in Turkish field strains. Conclusions: This study revealed that Turkish strains of BToV constitute a separate phylogenetic group and can be divided into two subgroups. In addition, BToV was found to be a common pathogen causing diarrhea in calves in Turkey, and it is a necessity to consider BToV in cases of diarrhea with unknown cause.

53-59

Authors: Da Eun Kim, Yeon Sil Hwang, Bo Yoon Chang, Dae Sung Kim, Hyoung Kwon Cho, Sung Yeon Kim

Objective: To investigate cosmetic potential of Syzygium aromaticum L. (S. aromaticum L.) and to determine its antioxidant and anti-wrinkling effects. Methods: Using high-performance liquid chromatography, eugenol component was quantitated. The antioxidant activity of S. aromaticum L. was analyzed by 2,2-diphenyl-1-picrylhydrazyl radical scavenging and superoxide dismutase like activities. To determine cell viability, elastase and matrix metalloproteinase-1 (MMP-1) activity, human dermal fibroblasts (HS68) were treated with S. aromaticum L. The inhibitory effect of S. aromaticum L. on tumor necrosis factor alpha induced MMPs expression in HS68 was analyzed by realtime-PCR. Results: The eugenol content was confirmed in S. aromaticum L. S. aromaticum L. was observed to have high 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity and superoxide dismutase like activity. S. aromaticum L. had no cytotoxicity against the HS68 and dose-dependently increased elastase inhibition. Moreover, S. aromaticum L. significantly decreased MMP-1 content and inhibited gene levels of MMP-1, MMP-2, MMP-3 and MMP-9. Conclusions: The findings suggest that S. aromaticum L. has great potential as a cosmeceutical ingredient with antioxidant and anti-wrinkling effects.

66-72

Authors: Jean Carlos Vencioneck Dutra, Judá Ben-Hur de Oliveira, Vanessa Silva dos Santos, Paula Roberta Costalonga Pereira, Jean Moisés Ferreira, Maria do Carmo Pimentel Batitucci

Objective: To evaluate the influence of fruiting phenological stage on total flavonoid content, antioxidant activity, and antiproliferative effects of Cereus jamacaru (C. jamacaru) (mandacaru) cladodes and fruit. Methods: Fruit and cladodes at vegetative and fruiting stage of C. jamacaru were collected. The fruit was dissected and bark, pulp, and seeds were separated. Vegetative and fruiting cladodes, together with bark, pulp, and seeds were used to obtain five hydroalcoholic extracts. The extracts were investigated for total flavonoid content, using AlCl3 colorimetric method, antioxidant activity by 2,2-diphenyl-1-picrylhydrazyl and 2,2’-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) radical scavenging capacity and Fe2+ ion chelating activity, and in vitro antiproliferative effects (sarcoma 180 cells) by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay. Results: The extract of C. jamacaru cladodes at the fruiting stage showed higher flavonoid content compared to the other extracts. Seed extracts showed the highest antioxidant activity in 2,2-diphenyl-1-picrylhydrazyl and 2,2’-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) assays, and the extract of cladodes at vegetative stage showed better antioxidant activity in Fe2+ ion chelating activity. The extract of fruiting cladodes promoted higher antiproliferative effects compared to the other extracts. Conclusions: These findings suggest that fruiting increases the content of flavonoids and antiproliferative effects of C. jamacaru cladodes. Data reinforce the potential use of C. jamacaru cladodes and fruits as natural antioxidants and potent anticancer agent.

80-84

Authors: Urarat Nanna, Jarinyaporn Naowaboot, Linda Chularojmontri, Pholawat Tingpej, Suvara Wattanapitayakul

Objective: To examine the effect of Brassica oleracea extract (BO) on impaired glucose and lipid homeostasis in high-fat diet (HFD)-induced obese mice. Methods: Obesity of ICR mice was induced by feeding a HFD (45 kcal% lard fat) for 16 weeks. During the last 8 weeks of study period, obese mice were additionally administered with BO (100 and 200 mg/kg/day). The metabolic parameters were determined. The gene expressions of hepatic lipogenesis were also studied. Results: After 8 weeks of treatment, BO (100 and 200 mg/kg) significantly reduced hyperglycemia and improved insulin sensitivity (P < 0.05). The serum lipid (total cholesterol, triglyceride, and non-esterified fatty acid) and hepatic triglyceride and non-esterified fatty acid were decreased (P < 0.05). The levels of insulin and leptin in serum were also decreased (P < 0.05). Moreover, the expressions of hepatic lipogenic genes including sterol regulatory element-binding protein 1c, fatty acid synthase, and acetyl-CoA carboxylase were decreased by BO treatment (P < 0.05). Conclusions: These results suggest that BO is a new therapeutic agent for improving the homeostasis of glucose and lipid in HFD-induced obese mice probably by suppression of lipogenic genes in liver tissue.

501-509

Authors: The Han Nguyen, Thi Huyen Nguyen, Van Minh Nguyen, Thi Lan Phuong Nguyen, Thi Van Anh Tran, Anh Duy Do, Sang Moo Kim

Objective: To investigate antidiabetic and antioxidant activities of the extract and fractions from Vietnamese red seaweed Laurencia dendroidea. Methods: The seaweed Laurencia dendroidea was extracted by using microwave-assisted extraction method in 80% methanol. The seaweed extract was then fractionated using different solvents (n-hexane, chloroform, ethyl acetate, butanol and water). These obtained fractions were evaluated for α -glucosidase inhibitory and antioxidant activities. Antioxidant activities were tested using DPPH, nitric oxide radical scavenging and metal chelating assays. The enzyme inhibition mode was determined using Lineweaver-Burk plot. For acidic and thermal stabilities, the ethyl acetate fraction was treated at pH 2.0 and 100 °C, respectively. The residual inhibitory activity of the fraction was calculated based on the initial inhibitory activity. For in vivo antidiabetic activity, mice were divided into four groups, including normal control, diabetic control, diabetic mice treated with ethyl acetate fraction and diabetic mice treated with gliclazide. Blood glucose level of treated mice during acute and prolonged treatments was measured. To evaluate the toxicity of the ethyl acetate fraction, the body weight changes and activities of liver function enzymes (aspartate transaminase, alanine transaminase and gamma-glutamyl transferase) were carried out. Results: The extract of Laurencia dendroidea showed strong α-glucosidase inhibitory and DPPH radical scavenging activities. Methanolic concentrations affected both α-glucosidase inhibitory and antioxidant activities. A 80% aqueous methanol was the suitable solvent for extraction of enzyme inhibitors and antioxidants. Among solvent fractions, ethyl acetate fraction had the highest inhibitory activities against α -glucosidase with a mixed type of inhibition and the strongest antioxidant activities, and was stable under acidic and thermal conditions. The ethyl acetate fraction treated diabetic mice significantly reduced blood glucose level compared with the diabetic control group (13.16 mmol/L vs. 22.75 mmol/L after 3 hours of treatment). Oral administration of ethyl acetate fraction did not exhibit toxicity at a dose of 100 mg/kg body weight as determined by body weight changes and liver biochemical parameters. Conclusions: Laurencia dendroidea could be a potential source for production of antidiabetic and antioxidative agents.

518-523

Authors: Isaac O Akefe, Ibrahim L Yusuf, Victoria A Adegoke

Objective: To investigate the antioxidant efficacy of oral orientin on 900 MHz radiofrequency-electromagnetic radiation-induced oxidative stress in mice. Methods: The mice were randomly allotted into 5 groups consisting of 7 mice each. The deionised water and radiofrequency electromagnetic radiations (RF-EMR) groups were administered with deionised water while orientin was administered to the RF-EMR + low dose of orientin group (10 mg/kg), RF-EMR + high dose of orientin group (20 mg/kg) and high dose of orientin group (20 mg/kg). All the groups except deionised water and high dose groups were exposed to 900 MHz radiofrequency-electromagnetic radiation for 28 consecutive days (1 h/day). Learning and memory was assessed via the step-down inhibitory avoidance task. Activities of lipid prexidation and antioxidant enzymes were measured using kits. Results: Radiofrequency electromagnetic radiation caused impairment in learning and memory and reduced activities of brain antioxidant enzymes, increased lipoperoxidation and corticosterone concentration as well as histopathological aberrations in the hippocampal tissues. Conversely, orientin alleviated learning and memory deficit, improved the activities of endogenous antioxidant enzymes and mitigated brain lipoperoxidation and neuronal degeneration in mice exposed to radiofrequency electromagnetic radiation. Conclusions: Orientin alleviates learning and memory impairment due to radiofrequency electromagnetic radiation in mice by improving antioxidant defence mechanism and may be considered as a promising therapeutic agent for improving the antioxidant system of people living in radiofrequency electromagnetic radiation-prone environment.

531-538

Authors: Adam Hermawan, Herwandhani Putri

Objective: To identify the potential target and mechanisms of hesperidin in MCF-7 estrogen receptor-positive breast cancer cells using bioinformatics approaches. Methods: Gene expression profiles were accessed from public database GSE85871. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis was carried out with Database for Annotation, Visualization and Integrated Discovery. The protein-protein interaction network was analyzed by STRING-DB and visualized by Cytoscape. Transcription factor regulatory networks were constructed from TRED, TRRUST, RegNetwork and visualized by Cytoscape. Drug association analysis was conducted by WebGestalt. Results: GO and KEGG pathway enrichment analysis revealed biological processes, cellular components and molecular functions that were related to cancer and estrogen signaling pathways. KEGG pathway enrichment analysis of the genes in transcription factor-differential expression genes regulatory network showed regulation of cancer, estrogen signaling pathways, epidermal growth factor receptor tyrosine kinase inhibitor resistance, and endocrine resistance. Moreover, drug association analysis revealed that hesperidin affected the expression of the same gene as raloxifene. Conclusions: Hesperidin targets estrogen receptor signaling in estrogen receptor-positive breast cancer cells. Results of this study could trace the molecular mechanism of hesperidin in estrogen receptor-positive breast cancer cells and integrative bioinformatics analysis could accelerate drug discovery and development.