Exploring the potential of dermal fibroblast conditioned medium on skin wound healing and anti-ageing

Skin ageing is associated with a decrease in collagen and delayed wound healing. The previous study has shown that supplementation of growth factors can enhance the production of extracellular matrix (ECM) and efficiency of wound healing. This study aimed to produce a Dermal Fibroblast Conditioned Medium (DFCM) and evaluate its potential on in vitro skin wound healing and anti-ageing properties. DFCM were obtained by culturing confluent human dermal fibroblasts (n=3) in keratinocyte-specific medium (DFCM-KM), fibroblast-specific medium (DFCM-FM) and fibroblast-specific medium with growth supplements (DFCM-GM). Protein concentration was determined by Bicinchoninic Acid (BCA) assay. To evaluate the wound healing and anti-ageing properties, human dermal fibroblasts were supplemented with three different DFCM. Fibroblasts cultured with F12:DMEM+10%FBS was used as a control. The growth rate of fibroblasts was evaluated by culturing cells until day 4. Migration rate was evaluated at 12 h after seeding. In vitro healing rate was evaluated via scratch assay. Anti-ageing potential of DFCM was evaluated via RT2 Profiler PCR Array. Concentration of total protein was found to be significantly higher in DFCM-GM (2914.79±150.67mg/mL) compared to DFCM-KM and DFCM-FM. Out of the tested 12 anti-ageing genes, fibroblasts supplemented with DFCM-GM demonstrated a significantly higher expression of WRN compared to other conditions. In conclusion, DFCM-GM contains more proteins and found superior in modulating wound healing and anti-ageing properties

Neuroprotective effect of phospholipase A2 from Malaysian Naja Sumatrana venom against H2O2-induced cell damage and apoptosis

Oxidative stress is one of the factors involved in the pathogenesis of several neurodegenerative diseases. It has been reported that a secretory phospholipase A2 known as A2-EPTX-NSm1a has lower cytotoxicity in neuronal cells compared to its crude Naja sumatrana venom. In this study, A2-EPTX-NSm1a was tested for its neuroprotective activity on human neuroblastoma cells (SH-SY5Y) differentiated into cholinergic neurons against oxidative stress induced by hydrogen peroxide (H2O2). H2O2 treatment alone increased the caspase-3 and caspase-8 activities, whereas pre-treatment with A2-EPTX-NSm1a reduced the activity of these apoptosis-associated proteins. Moreover, A2-EPTX-NSm1a protects the morphology and ultrastructure of differentiated SH-SY5Y cells in the presence of H2O2. Oxidative stress increased the number of small mitochondria. Further evaluation showed the size of mitochondria with a length below 0.25 µm in oxidative stress conditions is higher than the control group, suggesting mitochondria fragmentation. Pre-treatment with A2-EPTX-NSm1a attenuated the number of mitochondria in cells with H2O2 Furthermore, A2-EPTX-NSm1a altered the expression of several neuroprotein biomarkers of GDNF, IL-8, MCP-1, TIMP-1, and TNF-R1 in cells under oxidative stress induced by H2O2. These findings indicate that anti-apoptosis with mitochondria-related protection, anti-inflammatory effect, and promote expression of important markers for cell survival may underlie the neuroprotective effect of A2-EPTX-NSm1a in cholinergic rich human cells under oxidative stress, a vital role in the neuronal disorder

Identification of changes in the serum proteome of spontaneously hypertensive rats (SHR) following administration of aqueous extracts from ficus deltoidea / Nur Atiqah Haizum Abdullah

Ficus deltoidea or better known as Mas Cotek is an herbaceous plant found around the coast of Peninsular Malaysia especially in Terengganu and Kelantan. This plant is believed to have the capability in helping treat or controlling diseases including hypertension. Hypertension is a major disease that affects by many people all over the world. It is one of the main diseases that contribute to high rate of mortality. Even though modern medicine have been used to treat hypertension, some people still believe that traditional medicinal practices such as taking natural products along with healthy lifestyle will significantly improve hypertension. In this study, the capability of F. Deltoidea aqueous extracts in controlling hypertension were evaluated in vitro; using Angiotensin Converting Enzyme (ACE) inhibitory activity assay and through an in vivo study; by looking at the alteration of serum protein profiles in spontaneously hypertensive rats (SHR). Different parts of the plants obtained from two F.deltoidea varieties were used: fruits from the small-leaf variety (SF) and leaves from the big-leaf variety (BL). Changes in serum protein profiles were studied using proteomics techniques involving Sodium Dodecyl Sulphate- Polyacrylamide Gel Electrophoresis (SDS-PAGE), two-dimensional gel electrophoresis (2-DE), Matrix-assisted Laser Desorption / Ionization-Time of Flight / Time of Flight mass spectrometry (MALDI-TOF/TOF) and Surface-Enhanced Laser Desorption / Ionization-Time of Flight mass spectrometry (SELDI-TOF). Based on the results obtained, F. deltoidea showed positive effects towards the inhibition of ACE with the IC50 values of 0.13 and 0.18 mg/ml for SF and BL respectively. Thus, both samples were considered to be equally efficient as ACE inhibitory agent. Results had also demonstrated significant alterations in the serum protein profiles expressions of treated SHR rats as compared to normal and diseased rats (non-treated SHR). Seven serum proteins were confidently identified by MALDI-TOF/TOF which includes plasma retinol binding protein, serum albumin fragment, complement c3 precursor, haptoglobin, Apolipoprotein H, alpha 1 macroglobulin (A1M) and alpha 1 antiproteinase. All of these proteins showed possible association with hypertension in which A1M being the most related. SELDI-TOF univariate analysis revealed the alterations towards low molecular weight (LMW) serum proteins (2,500-40,000 kDa). Three protein peaks were altered following treatment with BL and seven with SF. SELDI-TOF multivariate analysis suggested several LMW serum proteins as potential biomarker candidates for hypertension; proteins with the m/z 33086 and 25706 (following SF treatment) and m/z 1617.5 and 5059.7 (following BL treatment). This study provides useful information that will greatly enhance the investigation of the exact mechanism of F. deltoidea in combating hypertension. These scientific evidences support the use of this plant as potential antihypertensive agent as claimed traditionally

Role of Olive Bioactive Compounds in Respiratory Diseases

Respiratory diseases recently became the leading cause of death worldwide, due to the emergence of COVID-19. The pathogenesis of respiratory diseases is centred around inflammation and oxidative stress. Plant-based alongside synthetic drugs were considered as therapeutics due to their proven nutraceutical value. One such example is the olive, which is a traditional symbol of the MedDiet. Olive bioactive compounds are enriched with antioxidant, anti-inflammatory, anticancer and antiviral properties. However, there are few studies relating to the beneficial effect of olive bioactive compounds on respiratory diseases. A vague understanding of its molecular action, dosage and bioavailability limits its usefulness for clinical trials about respiratory infections. Hence, our review aims to explore olive bioactive compound’s antioxidant, anti-inflammatory and antiviral properties in respiratory disease defence and treatment. Molecular insight into olive compounds’ potential for respiratory system protection against inflammation and ensuing infection is also presented. Olive bioactive compounds mainly protect the respiratory system by subsiding proinflammatory cytokines and oxidative stress

Effects of Hydroxytyrosol in Endothelial Functioning: A Comprehensive Review

Pharmacologists have been emphasizing and applying plant and herbal-based treatments in vascular diseases for decades now. Olives, for example, are a traditional symbol of the Mediterranean diet. Hydroxytyrosol is an olive-derived compound known for its antioxidant and cardioprotective effects. Acknowledging the merit of antioxidants in maintaining endothelial function warrants the application of hydroxytyrosol in endothelial dysfunction salvage and recovery. Endothelial dysfunction (ED) is an impairment of endothelial cells that adversely affects vascular homeostasis. Disturbance in endothelial functioning is a known precursor for atherosclerosis and, subsequently, coronary and peripheral artery disease. However, the effects of hydroxytyrosol on endothelial functioning were not extensively studied, limiting its value either as a nutraceutical supplement or in clinical trials. The action of hydroxytyrosol in endothelial functioning at a cellular and molecular level is gathered and summarized in this review. The favorable effects of hydroxytyrosol in the improvement of endothelial functioning from in vitro and in vivo studies were scrutinized. We conclude that hydroxytyrosol is capable to counteract oxidative stress, inflammation, vascular aging, and arterial stiffness; thus, it is beneficial to preserve endothelial function both in vitro and in vivo. Although not specifically for endothelial dysfunction, hydroxytyrosol safety and efficacy had been demonstrated via in vivo and clinical trials for cardiovascular-related studies

Isolation and characterization of A2-EPTX-Nsm1a, a secretory phospholipase A2 from Malaysian spitting cobra (naja sumatrana) venom

Phospholipase A2 (PLA2) toxins are one of the main toxin families found in snake venom. PLA2 toxins are associated with various detrimental effects, including neurotoxicity, myotoxicity,hemostatic disturbances, nephrotoxicity, edema, and inflammation. Although Naja sumatrana venom contains substantial quantities of PLA2 components, there is limited information on the function and activities of PLA2 toxins from the venom. In this study, a secretory PLA2 from the venom of Malaysian N. sumatrana, subsequently named A2-EPTX-Nsm1a, was isolated, purified, and characterized. A2-EPTX-Nsm1a was purified using a mass spectrometry guided approach and multiple chromatography steps. Based on LC-MSMS, A2-EPTX-Nsm1a was found to show high sequence similarity with PLA2 from venoms of other Naja species. The PLA2 activity of A2-EPTX-Nsm1 was inhibited by 4-BPB and EDTA. A2-EPTX-Nsm1a was significantly less cytotoxic in a neuroblastoma cell line (SH-SY5Y) compared to crude venom and did not show a concentration-dependent cytotoxic activity. To our knowledge, this is the first study that characterizes and investigates the cytotoxicity of an Asp49 PLA2 isolated from Malaysian N. sumatrana venom in a human neuroblastoma cell line

Neuroprotective effect of phospholipase A₂ from Malaysian<i> Naja sumatrana</i> venom against H₂O₂-induced cell damage and apoptosis

Oxidative stress is one of the factors involved in the pathogenesis of several neurodegenerative diseases. It has been reported that a secretory phospholipase A(2) known as A2-EPTX-NSm1a has lower cytotoxicity in neuronal cells compared to its crude Naja sumatrana venom. In this study, A2-EPTX-NSm1a was tested for its neuroprotective activity on human neuroblastoma cells (SH-SY5Y) differentiated into cholinergic neurons against oxidative stress induced by hydrogen peroxide (H(2)O(2)). H(2)O(2) treatment alone increased the caspase-3 and caspase-8 activities, whereas pre-treatment with A2-EPTX-NSm1a reduced the activity of these apoptosis-associated proteins. Moreover, A2-EPTX-NSm1a protects the morphology and ultrastructure of differentiated SH-SY5Y cells in the presence of H(2)O(2). Oxidative stress increased the number of small mitochondria. Further evaluation showed the size of mitochondria with a length below 0.25 µm in oxidative stress conditions is higher than the control group, suggesting mitochondria fragmentation. Pre-treatment with A2-EPTX-NSm1a attenuated the number of mitochondria in cells with H(2)O(2) Furthermore, A2-EPTX-NSm1a altered the expression of several neuroprotein biomarkers of GDNF, IL-8, MCP-1, TIMP-1, and TNF-R1 in cells under oxidative stress induced by H(2)O(2). These findings indicate that anti-apoptosis with mitochondria-related protection, anti-inflammatory effect, and promote expression of important markers for cell survival may underlie the neuroprotective effect of A2-EPTX-NSm1a in cholinergic rich human cells under oxidative stress, a vital role in the neuronal disorder