Asian Pacific Journal of Tropical Biomedicine

523-531

Authors: Xiu-Mei Bo, Ruo-Bing Yu, Sai-Jun Du, Rong-Li Zhang, Ling He

Objective: To identify the anti-depressive effect of ferulic acid (FA) in mice exposed to lipopolysaccharide (LPS) and explore its molecular mechanisms. Methods: The mice were divided into 5 groups as follows: Control, LPS, LPS + SP, LPS + FA, and LPS + FA + anisomycin. The LPS + FA and LPS + FA + anisomycin groups were administered with FA (100 mg/kg, i.p.) once daily continuously for 7 days, and the other groups received an equivalent volume of saline. On the 7th day, LPS (0.1 mg/mL, i.p.) was injected in all mice except the control group 30 min after FA or saline administration. The LPS + SP and LPS + FA + anisomycin groups were intravenously administered with SP600125 [c-Jun N-terminal kinase (JNK) inhibitor] (100 μL/ site, i.v.) and anisomycin (JNK activator) (100 μL/site, i.v.) 15 min before LPS, respectively. The depressive behaviors were assessed by open field test, sucrose preference test, and forced swimming test at 24 h post-LPS administration. Tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) levels in plasma were measured by ELISA. The levels of phospho-JNK, TNF-α, IL-1β, Bcl-2, Bax, cytochrome c and caspase-3 were evaluated by Western blotting. Results: FA alleviated depression symptoms caused by LPS in mice, including increasing sucrose water consumption in sucrose preference test and reducing the immobility time in forced swimming test. FA could inhibit upregulated levels of phospho-JNK, TNF-α, and IL-1β. FA also markedly decreased Bax, caspase-3, and cytochrome c, and increased Bcl-2 levels. Besides, SP600125 showed neuroprotective effect similar to FA which was attenuated by anisomycin. Conclusions: FA attenuates inflammation and apoptosis by inhibiting LPS-induced activation of JNK to alleviate depressionlike behaviors.

540-546

Authors: Shaza A Alyamani, Mayson H Alkhatib, Faiza Abdu

Objective: To evaluate the protective effect of the coconut oil nanoemulsion against methotrexate-induced hepatotoxicity and nephrotoxicity in Ehrlich ascites carcinoma-bearing Swiss albino mice. Methods: Forty mice were divided into four groups. Group I served as the untreated Ehrlich ascites carcinoma-bearing mice while Ehrlich ascites carcinoma-bearing mice in groups II-IV received an intraperitoneal injection of 0.2 mL/kg coconut oil nanoemulsion, 20 mg/kg methotrexate as well as 0.2 mL/kg coconut oil nanoemulsion mixed with 20 mg/kg methotrexate, respectively. The toxicities of the treatments were assessed by determining the complete blood count, performing the serum analysis for liver and kidney functions, evaluating the oxidative status and visualizing histological changes in the liver and kidney tissues. Results: Treatment with methotrexate and coconut oil nanoemulsion markedly diminished the liver parameters including aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, total protein, direct bilirubin and total bilirubin which were raised by methotrexate treatment (P < 0.05). Similarly, creatinine and blood urea nitrogen, as the indicators of kidney function, were dramatically lowered in the combination treatment group compared to the methotrexate group (P < 0.05). In addition, treatment with methotrexate and coconut oil nanoemulsion reduced the malondialdehyde and increased catalase, glutathione reductase and superoxide dismutase, in the liver and kidney tissues (P < 0.05). The treatment with methotrexate and coconut oil nanoemulsion reduced white blood cell count and increased the hemoglobin amount (P < 0.05), but did not cause any change in platelets and red blood cell count. Conclusions: Coconut oil nanoemulsion as a nanocarrier has great potential in reducing the adverse side effects induced by methotrexate.

555-562

Authors: Md. Nazim Uddin, Md. Abdus Samad, Md. Abu Zubair, Md. Zahurul Haque, Kanika Mitra, Tanzir Ahmed Khan, Md Amir Hossain, Ashikujaman Syed, Aklima Afroze

Objective: To investigate bioactive phytochemicals and antioxidant activities of Nymphaea nouchali and to explore its anticancer pathways by a network pharmacology approach. Methods: Using a spectrophotometer and high-performance liquid chromatography-diode array detector (HPLC-DAD), we quantified bioactive phytochemicals in methanolic extract of Nymphaea nouchali tuber. The extracts were investigated for in vitro antioxidant properties. Targets of these bioactive phytochemicals were predicted and anticancer-associated pathways were analyzed by a network pharmacology approach. Moreover, we identified the predicted genes associated with cancer pathways and the hub genes in the protein-protein interaction network of predicted genes. Results: Quantitative results indicated the total phenolics, total flavonoids, and total proanthocyanidins in the methanolic extract of Nymphaea nouchali tuber. HPLC-DAD analysis showed rutin (39.44 mg), catechin (39.20 mg), myricetin (30.77 mg), ellagic acid (11.05 mg), gallic acid (3.67 mg), vanillic acid (0.75 mg), rosmarinic acid (4.81 mg), p-coumaric acid (3.35 mg), and quercetin (0.90 mg) in 1 g of dry extract. The extract showed the radical scavenging activities of 2, 2-diphenyl-1-picrylhydrazyl, 2,2’-azino- bis (3-ethylbenzothiazoline-6-sulfonic acid) and N,N-dimethyl-p phenylenediamine. By using network pharmacology, we predicted 130 target genes associated with cancer pathways. The top hub genes (IL6, AKT1, EGFR, JUN, PTGS2, MAPK3, CASP3, and CXCL8) were also identified, which were associated with cancer pathways and interacted with bioactive phytochemicals of the methanolic extract of Nymphaea nouchali tuber. Conclusions: Our study provides insights into the mechanism of anticancer activities of the methanolic extract of Nymphaea nouchali tuber.

479-489

Authors: Esther O Aimofumeh, Godswill N Anyasor, Ijeoma Esiaba

Objective: To investigate the effect of Justicia secunda Vahl leaf fraction against acetaminophen-induced oxidative damage in the liver of rats. Methods: In vitro antioxidant assays were performed on Justicia secunda leaf fractions. Gas chromatography-mass spectrometry analytical method was done. Experimental animals were orally administered with 2 g/kg b.wt. acetaminophen, 100-500 mg/kg b.wt. Justicia secunda ethyl acetate leaf fraction (JSELF), and 100 mg/kg b.wt. silymarin. Blood and liver were collected to measure hepatic, oxidative stress, and membrane-bound phosphatase markers. Results: JSELF had significantly (P<0.05) high total antioxidant capacity and inhibition of lipid peroxidation. JSELF-treated animals had reduced plasma hepatic enzymes, serum C-reactive protein, and oxidized low-density lipoprotein while hepatic superoxide dismutase, catalase, and reduced glutathione levels were elevated compared with untreated control. Membrane-bound phosphatase activities were improved in JSELF-treated animals. GC-MS detected tentatively 7 antioxidants and 4 hepatoprotective compounds. Conclusions: JSELF could protect against oxidative stress and improve membrane-bound phosphatase activity in rats with acetaminophen-induced hepatic damage.

496-504

Authors: A Benkhaled, A Boudjelal, E Napoli, F Baali, G Ruberto

Objective: To evaluate chemical compositions, antioxidant and wound healing properties of Algerian Artemisia absinthium essential oil. Methods: The chemical composition of the essential oil from Artemisia absinthium was analyzed by a combination of GC-FID and GC/MS. The antioxidant capacities including the total antioxidant capacity, DPPH• and ABTS+• scavenging capacities were measured. The wound healing potential was assessed by the excision wound model of rats. The wounds were treated daily with an ointment prepared with two concentrations (5% and 10%) of Artemisia absinthium essential oil. The percentage of wound contraction was determined and wound healing was also evaluated by histological examination of the healed skin. Results: The main component of Artemisia absinthium essential oil was camphor (48%) followed by chamazulene (10%) that was responsible for the dark blue color of the oil. Artemisia absinthium essential oil exhibited moderate antioxidant activity compared with BHT and Trolox. All preparations showed significant effects on wound contraction and the ointment prepared with 10% of essential oil was effective as the reference drug Cicatryl. Conclusions: The essential oil of Artemisia absinthium shows moderate antioxidant activity. The 10% ointment enhances skin wound re-epithelialization and speeds up the healing process. The essential oil of Artemisia absinthium may be used as an alternative drug for wound healing.

516-522

Authors: Eun-Sun Hwang, Nhuan Do Thi

Objective: To determine the bioactive compounds of Orostachys japonicus (O. japonicus) (Maxim.) A. Berger extraction by different solvents (70% ethanol or water) and to evaluate the in vitro antioxidant and anti-inflammatory activities. Methods: Total polyphenol, flavonoid, and anthocyanin contents in O. japonicus extract were measured. Antioxidant activities were determined by DPPH, ABTS, and superoxide radical-scavenging ability assays. Anti-inflammatory activities were evaluated by nitric oxide production, tumor necrosis factor-α, and interleukin-6 expression techniques. Results: Extraction with 70% ethanol yielded the highest total polyphenol (60.03 mg/g dry weight) and flavonoid (55.50 mg/g dry weight) contents. The total anthocyanin contents in 70% ethanol and water extracts were 57.25 and 91.71 mg/g dry weight, respectively. The 70% ethanol extract also showed stronger antioxidant activity than the water extract. Antioxidant activity and reducing power increased with the increasing concentration of O. japonicus extract. O. japonicus extract at 0-400 μg/mL did not affect the growth of RAW 264.7 cells, whereas dose-dependent inhibition of nitric oxide, tumor necrosis factor-a, and interleukin-6 production was observed. Conclusions: O. japonicus inhibits oxidative and inflammatory reactions with potentially positive health-related effects.

433-441

Authors: Ali Akbar Asghari, Amin Mokhtari-Zaer, Saeed Niazmand, Kathleen Mc Entee, Maryam Mahmoudabady

Diabetes mellitus is a common metabolic disease with considerable morbidity and mortality. Untreated or improperly-treated diabetes can be associated with several long-term complications that necessitate an effective way to manage diabetes. Due to the side effects of synthetic glucose-lowering agents, alternative therapeutic modalities such as medicinal plants have attracted notable attention. Teucrium polium L. is a medicinal herb with antioxidant, antinociceptive, anti-inflammatory, hypolipidemic, hepatoprotective, and hypoglycemic properties. In vitro and in vivo studies have been conducted to characterize the anti-diabetic properties of Teucrium polium L. and its bioactive compounds. We conducted a literature study using Scopus, PubMed, and Google Scholar including the keywords “diabetes” and “Teucrium polium”. We also scanned all the references cited by the retrieved articles. According to this review, Teucrium polium administration displayed anti-diabetic effects by targeting different mechanisms and pathways, such as enhancement of insulin secretion and insulin level, improvement of oxidative damage, regeneration of pancreatic β-cells, and promotion of glucose uptake in muscle tissues by increasing GLUT-4 translocation as well as inhibiting α-amylase activity. Although Teucrium polium has been widely regarded as a traditional method, the pharmacological studies on anti-diabetic effects are not sufficient, most studies are either in-vivo or in-vitro. The preclinical and clinical studies are further required to confirm the efficacy of Teucrium polium.

452-459

Authors: Mustafa Hitit, Orhan Corum, Mehmet Ozbek, Kamil Uney, Ertugrul Terzi, Gokhan Arslan, AY Sonmez

Objective: To determine the anti-inflammatory effects of mucus obtained from different fish species on the carrageenan-induced acute paw edema in rats. Methods: Forty-two rats were randomly divided into seven groups. Acute paw edema was induced by 0.1 mL of 1% carrageenan, and a single dose of diclofenac and lyophilized mucus (25 mg/kg) of rainbow trout, brook trout, European sea bass, and gilthead sea bream were administered to rats through gastric gavage 1 h before carrageenan treatment. Rat paws were measured before and 1-4 h after carrageenan treatment. The mRNA expressions of cytokines (TNF-α, IL-1β, IL-6, IL-10, and TGF-β), antioxidant markers (catalase and superoxide dismutase), and COX-2 were investigated using quantitative polymerase chain reaction. The histopathological changes were evaluated by hematoxylin and eosin staining. Results: The inhibition percentage of carrageenan-induced paw edema by different fish mucus ranged from 52.46% to 74.86% at 4 h. Histopathological evaluation showed that all fish mucus diminished carrageenan-induced edema and inflammatory cell infiltration. The upregulation of IL-1β mRNA induced by carrageenan was decreased by the mucus of rainbow trout and gilthead sea bream while an increase in the expression of IL-6 mRNA was reduced by the mucus of rainbow trout, brook trout, and gilthead sea bream. In addition, the mRNA expression of superoxide dismutase was higher in the rainbow trout mucus group than the carrageenan group. Conclusions: Mucus obtained from different fish species may have anti-inflammatory effects.

470-478

Authors: Suphanthip Phusrisom, Laddawan Senggunprai, Auemduan Prawan, Sarinya Kongpetch, Upa Kukongviriyapan, Supawan Thawornchinsombut, Ronnachai Changsri, Veerapol Kukongviriyapan

Objective: To evaluate the immunomodulatory effects of rice bran hydrolysates on cultured immune cells and their underlying mechanism. Methods: Rice bran hydrolysates were prepared from pigmented rice (Oryza sativa L.) by hydrothermolysis and protease digestion. Rice bran hydrolysates were assayed for phenolic content and antioxidant activity. Cell proliferation of Jurkat, THP-1 and peripheral blood mononuclear cells (PBMC) was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Chemotaxis was evaluated by transwell chamber methods. Immunoadherence of THP-1 was performed on cultured human umbilical vein endothelial cells (HUVEC). Cytokine released from PBMC was measured by ELISA assay kits. Lymphocyte-mediated cytotoxicity was carried out on KKU-452 cells. Proteins associated with immunomodulation were analyzed by Western immunoblotting assay. Results: Rice bran hydrolysates were rich in phenolic compounds, such as ferulic acid, catechin, quercetin, and quercetin glycosides. Rice bran hydrolysates suppressed phytohemagglutinin (PHA)- stimulated proliferation of PBMC and Jurkat cells, chemotaxis of Jurkat and THP-1 cells, and immunoadherence of THP-1 on HUVEC cultured cells. The cellular mechanism of rice bran hydrolysates involved the activation of AMPK as well as suppression of mTOR, NF-κB and VCAM-1. Rice bran hydrolysates potentiated PBMC on the PHA-stimulated release of IL-2, TNF-α, and IL-4, and enhanced PHA-induced non-MHC-restricted cytotoxicity on KKU-452 cancer cells. Conclusions: The immunomodulatory effect of phytochemicals derived from rice bran hydrolysates suggests its therapeutic potential for further investigation.

1-10

Authors: Mohaddese Mahboubi, Mona Mahboubi

The protection of the liver as an essential organ in the body against oxidative stress and deleterious compounds has been the subject of recent investigations. Among different compounds, medicinal plants play an important role due to their hepatoprotective effects. Taraxacum officinale or “common dandelion” is a popular plant that has been traditionally used for its hepatoprotective effects. Currently, there are limited clinical studies on its hepatoprotective effects. The aim of this review article is to evaluate the hepatoprotective effects of dandelion and its mechanism of action. We reviewed literature up to July 2019 on “Taraxacum officinale” or “dandelion” and hepatoprotection. Currently available pharmacological studies indicate that dandelion extracts have hepatoprotective effects against chemical agents due to its antioxidant and anti-inflammatory activities. The anti-inflammatory effects of dandelion, the prebiotic effects of its oligofructans, inhibitory effects against the release of lipopolysaccharides and fasting induced adipose factor, digestive enzymes, and enhancing effects of lipogenesis, reduce lipid accumulation and liver inflammation, which directly or indirectly improve the liver functions. Given emerging evidence on hepatoprotective effects of dandelion, designing large human clinical studies is essential.

18-22

Authors: Ali Alkaladi, Haytham Ali, Aaser M Abdelazim, Mohamed Afifi, Mohammed Baeshen, Ammar AL-Farga

Objective: To investigate the molecular mechanisms of the anti-obese effect of raspberry ketone against high-fat diet fed rats. Methods: Fifty adult male rats were randomly assigned to receive a standard diet, a high fat diet, and the high-fat diet and 0.5%, 1% or 2% raspberry ketone. Body weight, biochemical parameters and gene expression of CCAAT enhancer-binding protein (C/EBP)δ, fatty acid synthase (FAS), acetyl CoA carboxylase (ACC), peroxisome proliferator-activated receptor alpha (PPAR-α), hormone-sensitive lipase (HSL) and hepatic carnitine palmitoyltransferase 1 A (CPT1A) were investigated. Results: Body weight, blood glucose, insulin, total lipids, triacylglycerols, total cholesterol and low-density lipoprotein cholesterol were increased in high-fat diet fed rats. These high fat diet-induced changes were attenuated by treatment with raspberry ketone. High-density lipoprotein cholesterol was decreased in highfat diet fed rats but increased in rats treated with raspberry ketone. Molecular investigations showed induction of gene expression of C/EBP-δ, FAS, ACC, CPT1A and inhibition of gene expression of PPAR-α and HSL in high-fat diet fed rats as compared with control. Raspberry ketone treament reversed these changes except CPT1A. Conclusions: Raspberry ketone can prevent obesity induced by a high-fat diet in rats by induction of the expression of enzymes, controlling lipolysis and fatty acids β oxidation as well as inhibition of gene expressions of adipogenic factors.

33-41

Authors: Yuling Ding, Chanipa Jiratchayamaethasakul, Junseong Kim, Eun-A Kim, Soo-Jin Heo, Seung-Hong Lee

Objective: To investigate the antioxidant and anti-melanogenesis activities of an ultrasonic extract of red sea cucumber, Stichopus japonicus, collected from Jeju Island. Methods: Antioxidant activity experiments were assessed by an electron spin resonance system and a cellular model of immortalized human keratinocytes (HaCaT) to determine its radical scavenging activity and protective effects against 2,2’-azobis (2-amidinopropane) dihydrochloride (AAPH)-induced oxidative stress. Anti-melanogenic activity of the ultrasonic extract of red sea cucumber was also examined using the melanoma cell model B16F10 and mushroom tyrosinase. Following the induction by α-melanocyte-stimulating hormone, the effects of the ultrasonic extract of red sea cucumber on intracellular tyrosinase activity, melanin content and the melanogenic protein expression of microphthalmia-associated transcription factor, tyrosinase, and tyrosinase-related proteins (TRP-1, and TRP-2) were examined. Results: The ultrasonic extract of red sea cucumber significantly scavenged 2,2-diphenyl-1-picrylhydrazyl and alkyl radicals [IC50: (0.924±0.035) and (0.327±0.006) mg/mL, respectively], as well as showed a protective effect against oxidative stress and attenuated generation of intracellular reactive oxygen species on AAPH- induced HaCaT cells, with no cytotoxicity (12.5-400 μg/mL). The ultrasonic extract of red sea cucumber also exhibited a tyrosinase inhibitory effect [IC50: (2.750±0.006) mg/mL]. On α-melanocyte-stimulating hormone-stimulated B16F10 melanoma cells, the ultrasonic extract of red sea cucumber (25-200 μg/mL) significantly inhibited not only melanin synthesis and tyrosinase activity, but also protein expressions of microphthalmia-associated transcriptional factor, tyrosinase, TRP-1, and TRP-2. Conclusions: The ultrasonic extract of red sea cucumber shows antioxidant and anti-melanogenic potential and may be a natural candidate for anti-aging as well as a whitening agent in the cosmeceuticals industry.

359-364

Authors: Marjan Zare, Abbas Rezaianzadeh, Hamidreza Tabatabaee, Hossain Faramarzi, Mohsen Aliakbarpour, Mostafa Ebrahimi

Objective: To determine the endemic values of cutaneous leishmaniasis in different cities of Fars province, Iran. Methods: Totally, 29 201 cases registered from 2010 to 2015 in Iranian Fars province were selected, and the endemic values of cutaneous leishmaniasis were determined by retrospective clusters derived from spatiotemporal permutation modeling on a time-series design. The accuracy of the values was assessed using receiver operating characteristic (ROC) curve. SPSS version 22, ArcGIS, and ITSM 2002 software tools were used for analysis. Results: Nine statistically significant retrospective clusters (P<0.05) resulted in finding seven significant and accurate endemic values (P<0.1). These valid endemic scores were generalized to the other 18 cities based on 6 different climates in the province. Conclusions: Retrospectively detected clusters with the help of ROC curve analysis could help determine cutaneous leishmaniasis endemic values which are essential for future prediction and prevention policies in the area.

373-380

Authors: Berween Mahmoud Elmahmoudy, Mai A Abd El Fattah, Mohamed F Elyamany, Laila A Rashed

Objective: To evaluate the antioxidant, immunomodulatory and anti-inflammatory activities of pyrrolidine dithiocarbamate and saxagliptin in rats with thioacetamide-induced ulcerative colitis. Methods: Animals were orally administered with a vehicle, sulfasalazine (500 mg/kg), pyrrolidine dithiocarbamate (100 mg/kg), and saxagliptin (10 mg/kg) for two weeks. Ulcerative colitis was induced by a single intrarectal instillation of thioacetamide on day 8. Colon samples were collected to assess mitogen-activated protein kinase (MAPK), phosphorylated extracellular signal-regulated kinase (ERK), cAMP response element-binding protein (CREB), interleukin-12 (IL-12), caspase-3, β-defensin, inducible nitric oxide synthase (iNOS) and glucagon like peptide-1 (GLP-1). Moreover, histopathological examination was performed. Results: Rats treated with thioacetamide caused increases in colonic MAPK, phosphorylated ERK, CREB, caspase-3, IL-12, β-defensin, iNOS, as well as decreases in body weight and GLP-1. In addition, distortion of colonic structure was found by histopathological examination. Pyrrolidine dithiocarbamate and saxagliptin mitigated colitis severity by improving body weight decrease and GLP-1, and reducing colonic MAPK, phosphorylated ERK, CREB, caspase-3, IL-12, β-defensin and iNOS. Conclusions: Pyrrolidine dithiocarbamate and saxagliptin are efficient against thioacetamide induced colitis through improving inflammatory and oxidative changes.

389-396

Authors: Sulistiyani , Syamsul Falah, Wulan Tri Wahyuni, Dimas Andrianto, Arthur Ario Lelono, Waras Nurcholis, Valeri Mossine, Mark Hannink

Objective: To fractionate and identify polyphenols from Guazuma ulmifolia Lam. leaves, and to explore their antioxidant, 5-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitory, and Nrf2 modulatory activities. Methods: The 1,1-diphenyl-2-picrylhydrazyl assay was used to evaluate the antioxidant activity of a polyphenolic fraction of the extract of Guazuma ulmifolia Lam. leaves. THP-1 gene reporter cell lines constructed with a transcriptional response element specific for Nrf2 and a minimal promoter for the firefly luciferase–green fluorescent protein transgene were used to determine the effect of the polyphenolic fraction on the Nrf2 signaling pathway. Furthermore, an assay of HMG-CoA reductase inhibitory activity was performed by using a commercial enzyme kit. Polyphenolic compounds were identified by liquid chromatography-tandem mass spectrometry. Results: The polyphenolic fraction showed fairly strong antioxidant activity [IC50 = (14.90 ± 4.70) μg/mL] and inhibited HMG-CoA reductase activity by 69.10%, which was slightly lower than that by pravastatin (84.37%) and quercetin (84.25%). Additionally, the polyphenolic fraction activated the Nrf2 antioxidant signaling pathway at 500 μg/mL. Eleven subfractions resulting from the column chromatography separation of the polyphenolic fraction also showed relatively strong antioxidant activities (IC50: 17.46μ217.14 μg/mL). The subfraction (F6) stimulated the Nrf2 signaling pathway and had HMG-CoA reductase inhibitory activity (65.43%). Moreover, the subfraction contained two main flavonoids: quercetin and quercimeritrin. Conclusions: The polyphenolic fraction of Guazuma ulmifolia could induce antioxidant genes via the Nrf2/antioxidant regulatory elements pathway, and is a promising candidate for an inhibitor of HMG-CoA reductase.