Asian Pacific Journal of Tropical Biomedicine

141-147

Authors: Shu-Ting Yang, Shu-Bai Liu

Cardiovascular diseases cause significant morbidity and mortality worldwide, incurring a major public health burden. Gastrodia elata Blume is a traditional Chinese herbal medicine that has been widely used to treat central nervous system and cardiovascular diseases. Gastrodin, as the major active component in Gastrodia elata Blume, can confer protection against cardiovascular diseases. In this review, we summarize the anti-inflammatory actions, anti-cardiac hypertrophy, anti-hypertension, anti-atherosclerosis, and angiogenic effects of gastrodin, as well as its protective effects on vascular cells and against myocardial ischemia-reperfusion injury. The medical potential of gastrodin in diabetes-related cardiovascular diseases is also discussed.

156-163

Authors: Renan Braga, Humberto Andrade, Ryldene Cruz, Mayara Maia, Carolina Lima, Anderson Santos, André Miranda, Allana Duarte, Marcus Scotti, Reinaldo Almeida, Damião Sousa

Objective: To evaluate the antinociceptive activity of perillyl acetate in mice and in silico simulations. Methods: The vehicle, perillyl acetate (100, 150 and/or 200 mg/ kg, i.p.), diazepam (2 mg/kg, i.p.) or morphine (6 mg/kg, i.p.) was administered to mice, respectively. Rotarod test, acetic acid-induced abdominal writhing, formalin-induced nociception, hot plate test, and tail-flick test were performed. Opioid receptors-involvement in perillyl acetate antinociceptive effect was also investigated. Results: Perillyl acetate did not affect the motor coordination of mice. However, it reduced the number of acetic acid-induced abdominal twitches and licking times in the formalin test. There was an increase of latency time in the tail-flick test of 30 and 60 minutes. Pretreatment with naloxone reversed the antinociceptive effect of perillyl acetate (200 mg/kg). In silico analysis demonstrated that perillyl acetate could bind to μ-opioid receptors. Conclusions: Perillyl acetate has antinociceptive effect at the spinal level in animal nociception models, without affecting the locomotor integrity and possibly through μ-opioid receptors. In silico studies have suggested that perillyl acetate can act as a μ-opioid receptor agonist.

175-184

Authors: Ranneh Yazan, Abu Bakar Mohd Fadzelly, Rahim Azlen-Che, Kassim Nur Kartinee, Stanslas Johnson, Teh Yuan-Han, Fadel Abdulmannan, S Ellulu Mohammed

Objective: To determine the lead bioactive compound in kernel extract of Mangifera pajang and its anti-cancer activity against human breast cancer cell lines with positive estrogen receptor (MCF-7). Methods: The methanolic extract of dried powder kernel of Mangifera pajang was exposed to column chromatography for isolation. The structural elucidation of the isolated compound was characterized using infrared, nuclear magnetic resonance, mass spectrometry. Furthermore, cytotoxicity, morphological changes, flow cytometry and cell cycle arrest analyses were performed to examine the mechanism of anti-proliferation and apoptosis induced by methyl gallate against MCF-7. Results: One compound was isolated from the methanolic extract of Mangifera pajang kernel and identified as methyl gallate. The flow cytometric results demonstrated induction of apoptosis in MCF-7 cells by three concentrations of methyl gallate. The cell cycle arrest showed a significant (P<0.05) decrease in cell progression at G2/M phase of MCF-7 after treatment with 100 μM of methyl gallate. The cell percentage of early and late apoptosis was significant at 10 and 100 μM of methyl gallate. Also, methyl gallate treatment induced up-regulation of reactive oxygen species levels in MCF-7 cells with a reduction in superoxide dismutase levels. Conclusions: These findings indicate that isolated methyl gallate from Mangifera pajang kernel extracts induces growth inhibition and apoptosis in MCF-7 cells via up-regulating oxidative stress pathway.

99-114

Authors: Rehab F Abdel-Rahman

Non-alcoholic fatty liver disease (NAFLD) denotes a spectrum of fatty liver disease in individuals without significant alcohol consumption. NAFLD is set to be the most common etiology of serious liver diseases in numerous nations when accompanied by obesity and type 2 diabetes. It is further histologically categorized into the non-alcoholic fatty liver (NAFL; steatosis without hepatocellular injury) and non-alcoholic steatohepatitis (NASH) which is characterized by the coexistence of hepatic steatosis and inflammation and is accompanied by hepatocyte injury (ballooning), either with or without fibrosis. NAFL is considered the benign and reversible stage arising from the excessive accumulation of triglycerides in hepatocytes. However, NASH is a more progressive stage of NAFLD, due to the increased risks of evolving more serious diseases such as cirrhosis, hepatocellular carcinoma. This concept, however, has been lately challenged by a hypothesis of multiple parallel hits of NAFLD, in which steatosis and NASH are separate entities rather than two points of the NAFLD spectrum, not only from a set of histological patterns but also from a pathophysiological perspective. The current review highlights the epidemiology and pathophysiology of NAFLD, and its progression towards steatohepatitis, with special focus on the novel imminent therapeutic approaches targeting the molecular aspects and the pathogenic pathways involved in the development, and progression of NAFLD.

124-131

Authors: Shivani S Wagh, Kalpesh R Patil, Umesh B Mahajan, Pradnya D Bagal, Avinash R Wadkar, Basavraj Bommanhalli, Prabhakar R Patil, Sameer N Goyal, Shreesh Ojha, Chandragouda R Patil

Objective: To compare the cardioprotective efficacy of equimolar doses (50 mM/kg, p.o.) of phloretin and genistein against doxorubicin-induced cardiotoxicity in rats. Methods: Cardiotoxicity was induced in rats by intraperitoneal injection of 6 mg/kg doxorubicin on alternative days till the cumulative dose reached 30 mg/kg. This study included four treatment groups of rats (n=6): the control group (0.5% carboxymethyl cellulose solution-treated), the doxorubicin- treated group (0.5% carboxymethyl cellulose solution along with doxorubicin), the genistein-treated group (50 mM/kg/day; p.o. along with doxorubicin) and phloretin-treated group (50 mM/kg/day; p.o. along with doxorubicin). On the 10th day of dosing, rats were anesthetized for recording ECG, mean arterial pressure, and left ventricular function. Oxidative stress, nitric oxide levels, and inflammatory cytokines were estimated in the cardiac tissue. Cardiac function parameters (creatine kinase MB, lactate dehydrogenase, aspartate aminotransferase, and alanine transaminase) were estimated in the serum samples. Results: Phloretin treatment inhibited doxorubicin-induced oxidative stress and also reduced nitric oxide levels in cardiac tissues of rats. Phloretin administration attenuated doxorubicin- induced alterations in hemodynamic parameters (heart rate, mean arterial blood pressure, and left ventricular function) and suppressed the expression of pro-inflammatory cytokines. The cardiac injury markers like creatine kinase MB, lactate dehydrogenase, aspartate aminotransferase, and alanine transaminase were reduced by both genistein and phloretin. All these effects of phloretin were more prominent than genistein. Conclusions: Phloretin offers cardioprotection that is comparable to genistein, a clinically validated cardioprotectant against doxorubicin-induced cardiotoxicity. Further studies are needed to confirm and establish the therapeutic utility of phloretin as a chemopreventive adjuvant to doxorubicin chemotherapy.

47-58

Authors: Sujatha M Hanumegowda, Chandramma Srinivasa, Ashwini Shivaiah, Manjula M Venkatappa, Ramesha Hanumanthappa, Rajesh Rangappa, Ramesh K Laxmaiah, Sathisha J Gonchigar, Devaraja Sannaningaiah

Objective: To explore the anticoagulant, antiplatelet and antioxidant activities of protein extract of kenaf seed (PEKS). Methods: Sodium dodecyl sulphate polyacrylamide gel electrophoresis and reverse-phase high-performance liquid chromatography techniques were employed for protein characterization. Antioxidant activity of PEKS was assessed using 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay. The protective effect of PEKS on sodium nitrite (NaNO2) induced oxidative stress was evaluated using the in vitro red blood cell model, while the effect of PEKS on diclofenac-induced oxidative stress was examined in vivo in rats. Platelet-rich plasma and platelet-poor plasma were used for anticoagulant and antiplatelet activities of PEKS. Results: PEKS revealed similar protein bands on SDS-PAGE under reduced and non-reduced conditions. Several acidic proteins were present in native PAGE. PEKS showed antioxidant properties by scavenging DPPH with an IC50 of 24.58 μg. PEKS exhibited a protective effect on NaNO2 induced oxidative stress in red blood cells by restoring the activity of stress markers. In addition, PEKS alleviated diclofenac-induced tissue damage of the liver, kidney, and small intestine. PEKS showed an anticoagulant effect in both in vivo and in vitro experiments by enhancing normal clotting time. PEKS did not affect prothrombin time but increase activated partial thromboplastin time. Furthermore, PEKS inhibited adenosine diphosphate and epinephrine-induced platelet aggregation. Conclusions: PEKS protects tissues from oxidative stress and exhibits antithrombotic activity.

69-77

Authors: Aishah E Albalawi, Norah A Althobaiti, Reem Hasaballah Alhasani, Sultan F Alnomasy

Objective: To assess the anti-tumor effects of Pistacia atlantica methanolic extract (PAME) compared with cyclophosphamide against Ehrlich solid tumors in mice. Methods: Swiss albino mice (n=40) were divided into five groups: normal control mice, mice with Ehrlich solid tumors treated with normal saline, mice with Ehrlich solid tumors treated with cyclophosphamide intraperitoneally once a day for 14 d, or 50 mg/kg or 100 mg/kg PAME orally once a day for 14 d. Tumor growth inhibition, body weight, tumor markers, liver and kidney enzymes, oxidative stress markers, antioxidant enzymes, tumor necrosis factor-alpha level (TNF-α), and apoptosis-regulatory gene expression were evaluated. Results: Treatment of mice bearing Ehrlich solid tumors with PAME at 50 and 100 mg/kg orally significantly decreased tumor volume, body weight, tumor markers, liver and kidney enzymes, oxidative stress markers and TNF-α level in comparison with mice with Ehrlich solid tumors receiving normal saline. whereas PAME at 50 and 100 mg/kg/day significantly elevated the level of antioxidant enzymes (P<0.05). Conclusions: Pistacia atlantica methanolic extract has potent antitumor activity in mice. Therefore, the extract might be considered as an alternative anticancer agent against tumors, however, additional studies especially in the clinical setting are required to confirm this finding.

87-98

Authors: Chiann Ying Yeoh, Andi Rifki Rosandy, Rozida Mohd Khalid, Yoke Kqueen Cheah

Objective: To elucidate the cytotoxic effect of the secondary metabolites of Barrientosiimonas humi (B. humi) on MCF-7 and MDA-MB-231 human breast cancer cells and its underlying mechanisms of action. Methods: The extract was obtained from the fermentation of B. humi and fractionation of the crude extract was conducted via column chromatography. Cytotoxicity of the B. humi extract was determined by using MTT assay and real-time cellular analysis. Morphological changes, cell cycle profiles, mode of cell death, and caspase expressions of control and treated breast cancer cells were determined. Results: The ethyl acetate extract isolated from B. humi was cytotoxic against MCF-7 and MDA-MB-231 cell lines. One of the dichloromethane (DCM) fractions, designated as DCM-F2, exhibited the strongest activity among all the fractions and thereby was selected for further studies. DCM-F2 had selective cytotoxicity on target cells by inducing apoptosis, particularly in the early stage, and cell cycle arrest. Treated cells caused inhibition of cell cycle progression at 72 h leading to a significant increase (P < 0.05) in the G0/G1 population. DCM-F2 treated MDA-MB-231 cells showed caspase-dependent apoptosis, whereas DCM-F2 treated MCF-7 cells showed a caspase-independent apoptosis pathway. Five compounds were successfully isolated from B. humi. Cyclo (Pro-Tyr) was the most cytotoxic and selective compound against MCF-7 cells. Conclusions: B. humi ethyl acetate extract exhibits significant cytotoxicity against MCF-7 and MDA-MB-231 cells via induction of apoptosis and cell cycle arrest.

495-503

Authors: Akbar Anaeigoudari

Medicinal plants and their ingredients have beneficial effects on human health. Nigella sativa is a herbal plant with multiple biological and pharmacological activities. Previous studies demonstrated the anti-inflammatory and antioxidant properties of Nigella sativa and its main constituent thymoquinone significantly contributes to the antidepressant and anti-nociception effects of this plant. It has been reported that thymoquinone may achieve its antidepressant effect by preventing the elimination of brain neurotransmitters affecting depression such as serotonin. The role of brain-derived neurotrophic factors in the antidepressant effects of thymoquinone has also been documented. Additionally, thymoquinone can attenuate pain by upregulation of intracellular signaling pathways related to nitric oxide and K+ATP channels. The present review summarizes the antidepressant and anti-nociceptive activity of Nigella sativa and its main constituent thymoquinone by searching literature on electronic databases such as PubMed, Web of Science, Scopus, and Google Scholar from the beginning of 2010 until the end of August 2022.

512-519

Authors: Qi-Feng Huang, Bo Wang, Yu-Qing Weng, Tang Deng, Li-Hua Li, Jin Qian, Qi Li, Kai-Wen Lin, Dong-Mei Sun, Shuang-Qin Xu, Hang-Fei Wang, Xin-Xin Wu, Yuan-Tian Sun, Xiao-Ran Liu

Objective: To evaluate the effect of midkine on lipopolysaccharide (LPS)-induced airway smooth muscle cells (ASMCs). Methods: LPS-stimulated acute lung injury model was used to analyze the effect of midkine on ASMCs in vitro. Recombinant midkine and midkine siRNA were used to investigate the role of Notch2 signaling pathway. Cell proliferation was assessed using Cell Counting Kit-8 assay. Additionally, apoptosis was measured by flow cytometry and protein and mRNA expression of midkine and Notch2 was assessed by Western blotting and qPCR, respectively. Immunofluorescence analysis was also conducted. Results: LPS increased the mRNA and protein expression of midkine and Notch2. Midkine silencing reduced LPS-induced midkine and Notch2 expression. In addition, midkine silencing further reduced the viability and increased apoptosis of ASMCs induced by LPS, which was attenuated by recombinant midkine. Conclusions: The midkine/Notch2 signaling pathway plays a regulatory role in ASMC proliferation and apoptosis in airway inflammation.

530-540

Authors: Smrutipragnya Samal, Rajesh Kumar Meher, Debasmita Dubey, Showkat Ahmad Mir, Binata Nayak, Mahesh Chandra Sahu, Pradeep Kumar Naik, Goutam Rath, Santosh Kumar Swain

Objective: To investigate the interaction of p53 with docetaxel and berberine and their anticancer activities against oral squamous cell carcinoma. Methods: The interaction between p53 with docetaxel and berberine was investigated and their mechanisms of action against oral squamous cell carcinoma were studied. Toxicity studies were performed to determine any toxic impact of the drugs on the vital organs of tested animals. Results: In silico results revealed the molecular interaction of docetaxel and berberine with p53 and the molecules were found to be potential p53 inducers. Docetaxel and berberine inhibited the proliferation of cancer cells in a concentration-dependent manner. Flow cytometry analysis revealed that docetaxel and berberine at IC50 concentrations upregulated the expression of p53 in oral squamous cell carcinoma cells, thus triggering apoptotic cell death. In addition, no toxicity was observed in the liver and kidney tissues of mice after docetaxel and berberine treatment. Conclusions: Docetaxel and berberine significantly suppressed the proliferation of oral cancer cells by activating p53 expression and causing apoptotic cell death. Both compounds can be potential agents for the treatment of oral cancer, with little to no toxicity at the tissue level.

453-465

Authors: Brian K. Beseni, Kolawole A. Olofinsan, Veronica F. Salau, Ochuko L. Erukainure, Md. Shahidul Islam

Objective: To explore the antioxidant and antidiabetic activities of Rhus longipes (R. longipes) leaf and stem bark aqueous infusions. Methods: R. longipes leaf and stem bark infusions were characterized via gas-chromatography mass-spectroscopy (GC-MS) analysis. In vitro antioxidant and carbohydrate and lipid digestive enzyme inhibitory activities of R. longipes infusions were determined. Additionally, the modulatory effects of R. longipes infusions on intestinal glucose absorption, muscle glucose uptake, and biomarkers of renal oxidative injury were evaluated. Molecular docking was performed to determine the binding affinities of the identified compounds from the leaf and stem bark infusions on carbohydrate and lipid digestive enzymes. Results: GC-MS analysis revealed the presence of several phytocompounds, including palmitoleic acid, octadecanamide, 24,25-dihydroxyvitamin D and L-ascorbic acid. The bark infusion had significantly higher total phenolic contents compared with the leaf infusion, with better DPPH scavenging [IC50: (10.50±1.03) ±g/mL] and ferric reducing [IC50: (9.85±0.32) ±g/mL] activities (P<0.05). Both R. longipes infusions at their highest concentrations significantly increased glucose uptake in yeast suspension and rat psoas muscle with marked suppression of glucose absorption in the rat jejunum (P<0.05). With no cytotoxicity on Vero cells, the infusions lowered lipid peroxidation, increased cellular reduced glutathione concentration, and the activities of superoxide dismutase and catalase in renal homogenate treated with FeSO4. Conclusions: R. longipes shows antioxidant and antidiabetic activities and could be a potential therapeutic candidate for diabetes.

475-482

Authors: Deepjyoti Dev, Ashish Sarkar, Bishnupada Roy

Objective: To evaluate the in vivo wound healing activity of Acanthus leucostachyus leaf extracts using an excision wound model in mice. Methods: Mice were divided into two groups of six animals in each group: the control group and the Acanthus leucostachyus extract-treated group. Healing potential was evaluated by determination of physical parameters (contraction rate, epithelialization period, and tensile strength), biochemical parameters (protein, DNA, and hydroxyproline content), the expression of growth factor and proinflammatory cytokines, as well as histological and ultrastructural observations. Results: Treatment with Acanthus leucostachyus leaf extracts markedly increased the rate of wound contraction, tensile strength, the concentrations of protein, DNA, and hydroxyproline, and the expression of growth factor, as well as promoted epithelialization, compared to the control. In addition, Acanthus leucostachyus leaf extracts significantly reduced the expression of pro-inflammatory cytokines. Histological and ultrastructural studies revealed the presence of thicker epithelial layer and smoother surface topography in the extract-treated group compared to the control. Conclusions: Acanthus leucostachyus leaf extracts show potent wound-healing activity and can be used as a wound healing agent.