Md. Mohibbullah

Moringa oleifera with promising neuronal survival and neurite outgrowth promoting potentials.

ETHNOPHARMACOLOGICAL RELEVANCE Moringa oleifera Lam. (Moringaceae) by virtue of its high nutritional as well as ethnomedical values has been gaining profound interest both in nutrition and medicinal research. The leaf of this plant is used in ayurvedic medicine to treat paralysis, nervous debility and other nerve disorders. In addition, research evidence also suggests the nootropic as well as neuroprotective roles of Moringa oleifera leaf in animal models. The aim of the present study was to evaluate the effect of Moringa oleifera leaf in the primary hippocampal neurons regarding its neurotrophic and neuroprotective properties. MATERIALS AND METHODS The primary culture of embryonic hippocampal neurons was incubated with the ethanol extract of Moringa oleifera leaf (MOE). After an indicated time, cultures were either stained directly with a lipophilic dye, DiO, or fixed and immunolabeled to visualize the neuronal morphology. Morphometric analyses for neurite maturation and synaptogenesis were performed using Image J software. Neuronal viability was evaluated using trypan blue exclusion and lactate dehydrogenase assays. RESULTS MOE promoted neurite outgrowth in a concentration-dependent manner with an optimal concentration of 30 μg/mL. As a very initial effect, MOE significantly promoted the earlier stages of neuronal differentiation. Subsequently, MOE significantly increased the number and length of dendrites, the length of axon, and the number and length of both dendrite and axonal branches, and eventually facilitated synaptogenesis. The β-carotene, one major compound of MOE, promoted neuritogensis, but the increase was not comparable with the effect of MOE. In addition, MOE supported neuronal survival by protecting neurons from naturally occurring cell death in vitro. CONCLUSIONS Our findings indicate that MOE promotes axodendritic maturation as well as provides neuroprotection suggesting a promising pharmacological importance of this nutritionally and ethnomedically important plant for the well-being of nervous system.

Gelidium amansii promotes dendritic spine morphology and synaptogenesis, and modulates NMDA receptor-mediated postsynaptic current

Neurotrophic factors are essential for the differentiation and maturation of developing neurons as well as providing survival support to the mature neurons. Moreover, therapeutically neurotrophic factors are promising to reconstruct partially damaged neuronal networks in neurodegenerative diseases. In the previous study, we reported that the ethanol extract of an edible marine alga, Gelidium amansii (GAE) had shown promising effects in the development and maturation of both axon and dendrites of hippocampal neurons. Here, we demonstrate that in primary culture of hippocampal neurons (1) GAE promotes a significant increase in the number of filopodia and dendritic spines; (2) promotes synaptogenesis; (3) enhances N-methyl-d-aspartic acid (NMDA) receptor recruitment; and (4) modulates NMDA-receptor-mediated postsynaptic current. Taken together these findings that GAE might be involved in both morphological and functional maturation of neurons suggest the possibility that GAE may constitute a promising candidate for novel compounds for the prevention and treatment of neurodegenerative diseases.

Differential Neuritogenic Activities of Two Edible Brown Macroalgae, Undaria pinnatifida and Saccharina japonica

Undaria pinnatifida (Harvey) Suringar and Saccharina japonica Areschoug are two common seaweeds, and both are known to have numerous pharmacological properties that include neuroprotective effects. In a previous study, we found that the ethanol extracts of U. pinnatifida (UPE) and S. japonica (SJE) had neurite promoting activities on developing hippocampal neurons. In the present study, we studied and compared the effects of UPE and SJE on neuronal maturation. Both UPE and SJE promoted neurite outgrowth in a dose-dependent manner with optimal concentrations of 5 and 15 μg/mL, respectively. Initial neuronal differentiation was significantly promoted by UPE and SJE. Subsequently, treatment with both increased indices of axonal and dendritic cytoarchitecture, such as, the numbers and lengths of primary processes, although only UPE had a significant effect on branching frequencies. In addition, UPE and SJE showed no evidence of cytotoxicity, rather they protected neurons from naturally occurring death in vitro. These results indicate that UPE and SJE promote axodendritic maturation and neuronal survival and suggest that these algal extracts, especially UPE, have beneficial effects on the nervous system.

The neuritogenic and synaptogenic effects of the ethanolic extract of radix Puerariae in cultured rat hippocampal neurons.

ETHNOPHARMACOLOGICAL RELEVANCE Radix Puerariae, the root of Pueraria montana var. lobata (Willd.) Sanjappa & Pradeep, is used in Korean traditional medicine to treat neuronal disorders including Parkinsons disease, and its active constituent, puerarin has been reported to have a neuroprotective effect in experimental models of Parkinsons and Alzheimers disease. AIMS OF THE STUDY To investigate the neurotrophic effects of these ethnomedicines on the development of central nervous system neurons and the molecular bases of these activities. MATERIALS AND METHODS Rat embryonic (E19) brain neurons were cultured in the absence or presence of the ethanolic extract of Radix Puerariae (RPE) or puerarin. At predetermined times, cells were fixed and immunostained to visualize neuronal morphologies, or lysed for protein harvesting. Morphometric analyses of neurite outgrowths and synaptogenesis were performed using Image J software. RPE or puerarin-mediated changes in the protein profiles of cultured neurons were assessed by MALDI-TOF-MS/PMF and measuring immunofluorescent intensities. RESULTS RPE and puerarin alone promoted maximum neurite outgrowths at concentrations of 1µg/ml and 5µM, respectively. At these optimal concentrations, RPE and puerarin provided neurotrophic support by promoting axo-dendritic arbors and synapse formation in cultured neurons. Proteomic study revealed that RPE and puerarin both up-regulated a number of proteins, including dynein light chain 2 (DLC2) and elongation factor 2 (EF2), which are associated with neuritogenesis and synaptic potentiation, respectively. Immunofluorescence intensity measurements confirmed the expressions of the DLC2 and Dync1h1 subunits of dynein in RPE or puerarin treated hippocampal neurons were up-regulated when RPE or puerarin induced changes in neuronal cytoarchitecture. CONCLUSIONS Our study demonstrates that RPE and puerarin should be considered potentially valuable preventative therapeutics for brain disorders due to their abilities to promote the neuronal cytoarchitecture and the synaptic functionality, which are possibly associated with dynein-dependent regulation of cytoskeletal structures and up-regulation of translation machinery.

Radix Puerariae modulates glutamatergic synaptic architecture and potentiates functional synaptic plasticity in primary hippocampal neurons

ETHNOPHARMACOLOGICAL RELEVANCE Neurologic disorders are frequently characterized by synaptic pathology, including abnormal density and morphology of dendritic spines, synapse loss, and aberrant synaptic signaling and plasticity. Therefore, to promote and/or protect synapses by the use of natural molecules capable of modulating neurodevelopmental events, such as, spinogenesis and synaptic plasticity, could offer a preventive and curative strategy for nervous disorders associated with synaptic pathology. Radix Puerariae, the root of Pueraria monatana var. lobata (Willd.) Sanjappa&Pradeep, is a Chinese ethnomedicine, traditionally used for the treatment of memory-related nervous disorders including Alzheimers disease. In the previous study, we showed that the ethanolic extracts of Radix Puerariae (RPE) and its prime constituent, puerarin induced neuritogenesis and synapse formation in cultured hippocampal neurons, and thus could improve memory functions. AIMS OF THE STUDY In the present study, we specifically investigated the abilities of RPE and puerarin to improve memory-related brain disorders through modulating synaptic maturation and functional potentiation. MATERIALS AND METHODS Rat embryonic (E19) brain neurons were cultured in the absence or presence of RPE or puerarin. At predetermined times, cells were live-stained with DiO or fixed and immunostained to visualize neuronal morphologies, or lysed for protein harvesting. Morphometric analyses of dendritic spines and synaptogenesis were performed using Image J software. Functional pre- and postsynaptic plasticity was measured by FM1-43 staining and whole-cell patch clamping, respectively. RPE or puerarin-mediated changes in actin-related protein 2 were assessed by Western blotting. Neuronal survivals were measured using propidium iodide exclusion assay. RESULTS RPE and puerarin both: (1) promoted a significant increase in the numbers, and maturation, of dendritic spines; (2) modulated the formation of glutamatergic synapses; (3) potentiated synaptic transmission by increasing the sizes of reserve vesicle pools at presynaptic terminals; (4) enhanced NMDA receptor-mediated postsynaptic currents, and (5) increased cell viability against naturally occurring cell death. Moreover, upregulation of actin-related protein 2 (ARP2) in RPE and puerarin treated brain neurons suggest that RPE and puerarin induced synaptic plasticity might be associated, at least in part, with ARP2-mediated actin-dependent regulation of spinogenesis. CONCLUSIONS Our findings indicate that RPE and puerarin might play a substantial role in the morphological and functional maturation of brain neurons and suggest that RPE and puerarin are potentially valuable preventative therapeutics for memory-related nervous disorders.

Effect of superheated steam roasting with hot smoking treatment on improving physicochemical properties of the adductor muscle of pen shell (Atrina pectinate)

Abstract The adductor muscle of the pen shell Atrina pectinata (AMPS) is a popular protein‐enriched food item in Asian Pacific countries, and has only been marketed in the frozen condition, as a result browning and decreased sensory attributes occur. To overcome these problems, superheated steam roasting (at 270°C for 4 min) combined with the hot smoke (10 min) using a selective Oak sawdust was employed to develop a new AMPS product yielding high physicochemical properties during storage periods (0–13 days) especially at 10°C. The processed AMPS showed high sensory preferences because of good odor, color, and textural properties. It also significantly inhibited bacterial growth, volatile basic nitrogen, thiobarbituric acid‐reactive species, and pH changes, and eventually possessed higher nutritional composition with low trimethylamine N‐oxide level. Results indicate that saturated steam allows AMPS at good physicochemical conditions, whereas hot smoke‐derived aroma compounds prolong its shelf life through antioxidant and antimicrobial activities.