Asian Pacific Journal of Tropical Biomedicine

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.

411-420

Authors: William Yousseu Nana, Ernest Allah Hoki Kwenteh, Gilbert Ateufack, Eric Gonzal Tsafack, Stephanie Flore Djuichou Nguemnang, Zenab Linda Fagni Njoya, Albert Donatien Atsamo, Vanessa Mba Matah Marthe, Carine Flore Adjouzem, Yacine Karelle Madjo Kouam, Elvira Ngoufack Azanze, Marius Mbiantcha

Objective: To evaluate the anti-arthritic effect of aqueous and methanolic extracts of Distemonanthus benthamianus. Methods: Monoarthritis was induced by an injection of 0.3 mL zymosan A (0.9% NaCl, v/v) in the right posterior knee joints of rats. Then, joint diameter and pain threshold were determined. Polyarthritis was induced by an intracaudal injection of complete Freund’s adjuvant and rats were treated from day 14 post 1st complete Freund’s adjuvant injection until 28 day. The clinical, hematological, biochemical and oxidative stress parameters were evaluated. In addition, histological analysis of the knee joint was perfomed in both tests. Results: The aqueous and methanolic extracts of Distemonanthus benthamianus at a dose of 500 mg/kg ameliorated zymosan A-induced monoarthritis, as evidenced by reduced joint diameter, increased pain threshold, as well as improved joint architecture. In addition, both extracts of Distemonanthus benthamianus markedly increased body weight and pain threshold, while reducing paw edema in polyarthritic rats. They also led to a marked decrease in platelets and white blood cells (P<0.05), as well as a significant increase in red blood cells, hemoglobin and hematocrit (P<0.05). The aqueous and methanolic extracts of Distemonanthus benthamianus significantly reduced alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase activities, while increasing serum protein levels (P<0.05) with no significant variation in creatinine level. Moreover, both extracts increased catalase and glutathione activities (P<0.05), and inhibited malondialdehyde and nitric oxide production (P<0.01 and P<0.001) in the liver and kidneys. Histological analysis of the joints showed that both extracts triggered tissue reparation. Conclusions: Distemonanthus benthamianus could be used as a potential candidate in the treatment of rheumatoid arthritis.

430-436

Authors: Hyun Ji Eo, Da Som Kim, Gwang Hun Park

Objective: To elucidate the potential anti-inflammatory mechanisms of Rhamnus crenata leaf extracts using RAW264.7 cells. Methods: We used 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide assay to measure cell viability. Nitric oxide (NO) production was measured using Griess reagent. Western blotting and RT-PCR assays were carried out for analyzing the protein and gene expressions of pro-inflammatory mediators, respectively. Moreover, PD98059 (ERK1/2 inhibitor), SB203580 (p38 inhibitor), SP600125 (JNK inhibitor), and BAY11-7082 (NF-κB inhibitor) were used to evaluate the anti-inflammatory mechanism of Rhamnus crenata leaf extract. Results: Rhamnus crenata leaf extracts significantly inhibited the production of the pro-inflammatory mediators such as NO, iNOS, COX-2, IL-1β, and TNF-α in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. Rhamnus crenata leaf extracts also suppressed LPS-induced degradation of IκB-α and nuclear accumulation of p65, which resulted in the inhibition of NF-κB activation in RAW264.7 cells. Additionally, the extracts attenuated the phosphorylation of p38, ERK1/2, and JNK in LPS-stimulated RAW264.7 cells. Moreover, HO-1 expression induced by Rhamnus crenata leaf extracts was significantly downregulated by SB230580, PD98059, SP600125 and BAY11-7082. Conclusions: Rhamnus crenata leaf extract may upregulate HO-1 expression through inhibition of p38, ERK1/2, and NF-κB activation, which may contribute to the anti-inflammatory activity of the extracts. Rhamnus crenata leaf extracts may have great potential for the development of anti-inflammatory drugs to treat acute and chronic inflammatory diseases.

446-452

Authors: Fairouz Sioud, Mouna Maatouk, Imen Mokdad Bzeouich, Leila Chekir Ghedira, Soumaya Kilani-Jaziri

Objective: To evaluate the effects of phenolic acids (caffeic, ferulic, and coumaric acids) and flavones (luteolin and apigenin) on the proliferation and melanogenesis in murine melanoma B16-F10 cells. Methods: Cell proliferation was determined after 24 and 48 hours of incubation using MTT assay. The effects of these tested compounds on cell cycle progression were analyzed by flow cytometry. Moreover, the melanin content and tyrosinase activity were measured spectrophotometrically at 475 nm. Results: Luteolin and apigenin exhibited significant anti-proliferative activity against B16-F10 cells, while caffeic, ferulic, and coumaric acids induced slight inhibition after 24 and 48 hours of incubation. The tested compounds disturbed cell cycle progression of B16-F10, by a subsequent decrease in G1 and arrested cycle progression in either G1/S or G2/M phase. Furthermore, apigenin provoked an increase in melanin content of B16-F10 cells. In contrast, luteolin, caffeic, ferulic and coumaric acids induced a decrease in melanin content of B16-F10 cells by inhibiting tyrosinase activity. Conclusions: These active polyphenols may be used as skin whitening agents or natural tanning agents to treat skin pigmentation disorders.