Sushruta Koppula

The role of free radicals in the aging brain and Parkinson's Disease: convergence and parallelism.

Free radical production and their targeted action on biomolecules have roles in aging and age-related disorders such as Parkinson’s disease (PD). There is an age-associated increase in oxidative damage to the brain, and aging is considered a risk factor for PD. Dopaminergic neurons show linear fallout of 5–10% per decade with aging; however, the rate and intensity of neuronal loss in patients with PD is more marked than that of aging. Here, we enumerate the common link between aging and PD at the cellular level with special reference to oxidative damage caused by free radicals. Oxidative damage includes mitochondrial dysfunction, dopamine auto-oxidation, α-synuclein aggregation, glial cell activation, alterations in calcium signaling, and excess free iron. Moreover, neurons encounter more oxidative stress as a counteracting mechanism with advancing age does not function properly. Alterations in transcriptional activity of various pathways, including nuclear factor erythroid 2-related factor 2, glycogen synthase kinase 3β, mitogen activated protein kinase, nuclear factor kappa B, and reduced activity of superoxide dismutase, catalase and glutathione with aging might be correlated with the increased incidence of PD.

Inhibitors of Microglial Neurotoxicity: Focus on Natural Products

Microglial cells play a dual role in the central nervous system as they have both neurotoxic and neuroprotective effects. Uncontrolled and excessive activation of microglia often contributes to inflammation-mediated neurodegeneration. Recently, much attention has been paid to therapeutic strategies aimed at inhibiting neurotoxic microglial activation. Pharmacological inhibitors of microglial activation are emerging as a result of such endeavors. In this review, natural products-based inhibitors of microglial activation will be reviewed. Potential neuroprotective activity of these compounds will also be discussed. Future works should focus on the discovery of novel drug targets that specifically mediate microglial neurotoxicity rather than neuroprotection. Development of new drugs based on these targets may require a better understanding of microglial biology and neuroinflammation at the molecular, cellular, and systems levels.

Protective effects of Gastrodia elata Blume on MPP+-induced cytotoxicity in human dopaminergic SH-SY5Y cells.

AIM OF THE STUDY Gastrodia elata (GE) Blume (Orchidaceae) has been traditionally used as a folk medicine in Oriental countries since centuries for their variety of therapeutic benefits. This study is an attempt to investigate the protective effects of GE extract against MPP(+)-induced cytotoxicity in human dopaminergic SH-SY5Y cells and explore the neuroprotective mechanisms involved. MATERIALS AND METHODS Human dopaminergic SH-SY5Y cells were used to demonstrate the protective effects of GE against multiple parameters such as MPP(+)-induced cell viability, oxidative damage, expression of Bcl-2 and Bax, caspase-3 and poly(ADP-ribose) polymerase proteolysis. RESULTS GE effectively attenuated the cytotoxicity and improved cell viability in a dose-dependent manner. GE was effective in inhibiting both, the increased production of reactive oxygen species (ROS) and increase in Bax/Bcl-2 ratio, cleaved caspase-3 and PARP proteolysis. CONCLUSION Data from this study suggests the protective effects of GE on MPP(+)-induced cytotoxicity in dopaminergic cells, which may be ascribed to its significant anti-oxidative and anti-apoptotic properties, thus, GE might prove to be a valuable therapeutic agent for the treatment of various neurodegenerative diseases including progressive Parkinsons disease (PD).

Regulation of microglia activity by glaucocalyxin-A: attenuation of lipopolysaccharide-stimulated neuroinflammation through NF-κB and p38 MAPK signaling pathways.

Microglial cells are the resident macrophages and intrinsic arm of the central nervous system innate immune defense. Microglial cells become activated in response to injury, infection, environmental toxins, and other stimuli that threaten neuronal survival. Therefore, regulating microglial activation may have therapeutic benefits that lead to alleviating the progression of inflammatory-mediated neurodegeneration. In the present study, we investigated the effect of glaucocalyxin A (GLA) isolated from Rabdosia japonica on the production of pro-inflammatory mediators in lipopolysaccharide (LPS)-stimulated primary microglia and BV-2 cells. GLA significantly inhibited LPS-induced production of nitric oxide and reversed the morphological changes in primary microglia. Further, GLA suppressed expression of inducible nitric oxide synthase and cyclooxygenase-2 dose-dependently at the mRNA and protein levels. The production of proinflammatory cytokines such as tumor necrosis factor-α, interleukin-1β (IL)-1β, and IL-6 were inhibited by suppressing their transcriptional activity. Furthermore, GLA suppressed nuclear factor-κB activation by blocking degradation of IκB-α and inhibited the induction of lipocalin-2 expression in LPS-stimulated BV-2 cells. Mechanistic study revealed that the inhibitory effects of GLA were accompanied by blocking the p38 mitogen activated protein kinase signaling pathway in activated microglia. In conclusion, given that microglial activation contributes to the pathogenesis of neurodegenerative diseases, GLA could be developed as a potential therapeutic agent for treating microglia-mediated neuroinflammatory diseases.

The Role of Bioactive Compounds on the Promotion of Neurite Outgrowth

Neurite loss is one of the cardinal features of neuronal injury. Apart from neuroprotection, reorganization of the lost neuronal network in the injured brain is necessary for the restoration of normal physiological functions. Neuritogenic activity of endogenous molecules in the brain such as nerve growth factor is well documented and supported by scientific studies which show innumerable compounds having neurite outgrowth activity from natural sources. Since the damaged brain lacks the reconstructive capacity, more efforts in research are focused on the identification of compounds that promote the reformation of neuronal networks. An abundancy of natural resources along with the corresponding activity profiles have shown promising results in the field of neuroscience. Recently, importance has also been placed on understanding neurite formation by natural products in relation to neuronal injury. Arrays of natural herbal products having plentiful active constituents have been found to enhance neurite outgrowth. They act synergistically with neurotrophic factors to promote neuritogenesis in the diseased brain. Therefore use of natural products for neuroregeneration provides new insights in drug development for treating neuronal injury. In this study, various compounds from natural sources with potential neurite outgrowth activity are reviewed in experimental models.

Nuclear factor erythroid 2-related factor 2 signaling in Parkinson disease: a promising multi therapeutic target against oxidative stress, neuroinflammation and cell death.

Parkinsons disease (PD) is the second most common progressive neurodegenerative disorder with increased oxidative stress as central component. Till date, treatments related to PD are based on restoring dopamine either by targeting neurotransmitter and/or at receptor levels. These therapeutic approaches try to repair damage but do not address the underlying processes such as oxidative stress and neuroinflammation that contribute to cell death. The central nervous system maintains a robust antioxidant defense mechanism consisting of several cytoprotective genes and enzymes whose expression is controlled by antioxidant response element (ARE) which further depends on activation of nuclear factor erythroid 2-related factor 2 (Nrf2). In response to oxidative or electrophilic stress transcription factor Nrf2 binds to ARE and rescues the cells from oxidative stress and neuroinflammation. Recently, Nrf2 has been utilized as a drug target and some agents are currently under clinical trial. Owing to the potential role of Nrf2 in counteracting oxidative stress and neuroinflammation seen in PD, here we have focused on the molecular mechanism of the Nrf2/ARE antioxidant defense pathway in PD. Further, we also summarize published reports on the potential inducers of Nrf2 that demonstrate neuroprotective effects in experimental models of PD with possible future strategies to increase the transcriptional level of Nrf2 as a therapeutic strategy to provide neuroprotection of damaged dopaminergic neurons in PD.

Cognitive Enhancing Effects of Alpha Asarone in Amnesic Mice by Influencing Cholinergic and Antioxidant Defense Mechanisms

The effect of α-asarone on impairment of cognitive performance caused by amnesic drug scopolamine was investigated. Treatment with α-asarone attenuated scopolamine-induced cognitive deficits as evaluated by passive avoidance and Y-maze test. Administration of α-asarone for 15 d improved memory and cognitive function as indicated by an increase in transfer latency time and spontaneous alternation in passive avoidance and the Y-maze test respectively. To understand the action of α-asarone, the levels of acetylcholinesterase (AChE), malondialdehyde (MDA), and superoxide dismutase (SOD) in the hippocampus (Hippo) and cerebral cortex (CC) of scopolamine-induced amnesic mice were evaluated. The mice treated with Scopolamine showed increased activity of AChE, MDA and SOD levels in both the Hippo and the CC area. Treatment with α-asarone attenuated the increased activity of AChE and normalized the MDA and SOD levels in the Hippo and the CC area in the scopolamine treated amnesic mice. These results suggest that α-asarone has a beneficial effect in cognitive impairment induced by dysfunction of cholinergic system in brain through inhibition of AChE activity and by influencing the antioxidant defense mechanism.

Protective effect of Chrysanthemum indicum Linne against 1-methyl-4-phenylpridinium ion and lipopolysaccharide-induced cytotoxicity in cellular model of Parkinson’s disease

Chrysanthemum indicum Linn. (CI) has been used in Oriental medicine for several centuries. In the present study, the effect of CI extract was evaluated against 1-methyl-4-phenylpridinium ion (MPP(+))-induced damage in SH-SY5Y cells and lipopolysaccharide (LPS)-stimulated BV-2 microglial cells. Cell viability, oxidative damage, reactive oxygen species, expression of Bcl-2/Bax, and poly (ADP-ribose) polymerase (PARP) proteolysis were evaluated using SH-SY5Y cells. Production of iNOS, prostaglandin E(2), and pro-inflammatory cytokines like tumor necrosis factor (TNF)-α, interleukin (IL)-1β, interleukin (IL)-6, expression of cyclooxygenase type-2 (COX-2) and type-1 (COX-1) were examined in activated BV-2 microglia. At 1, 10 and 100 μg, CI inhibited cell loss, decreased the reactive oxygen species production, regulated the Bax/Bcl-2 ratio and inhibited PARP proteolysis in MPP(+)-induced SH-SY5Y cells. Furthermore, CI suppressed the production of prostaglandin E(2,) expression of cyclooxygenase type-2 (COX-2), blocked IκB-α degradation and activation of NF-κB p65 in BV-2 cells in a dose-dependent manner. The molecular mechanisms involved by CI might involve its inhibitory actions both on neuronal apoptosis and neuroinflammatory NF-κB/IκB-α signaling pathway. The present investigation scientifically supports the long history and safe usage of CI as an important functional food with potential benefits in ameliorating deleterious conditions seen in PD.

Modulation of LPS-stimulated neuroinflammation in BV-2 microglia by Gastrodia elata: 4-hydroxybenzyl alcohol is the bioactive candidate.

ETHNOPHARMACOLOGICAL RELEVANCE Gastrodia elata Blume (Orchidaceae, GE) a traditional plant in Oriental countries is known for its enormous benefits to treat headaches, dizziness, vertigo and convulsive illnesses. In the present study, the ethnopharmacological role of GE in neuroinflammation mediated by activated microglia and the mechanisms underlying were reported. MATERIALS AND METHODS BV-2 microglia activated by lipopolysaccharide (LPS) was employed and the effects of GE on corresponding neuroinflammatory parameters were assessed. RESULTS GE extract inhibited LPS-stimulated production of inflammatory cytokines and down regulated the c-Jun NH(2)-Terminal Kinase (JNK) and nuclear factor-kappa B (NF-κB) signaling pathways, which are known to be involved in neuroinflammation. Further, inhibition of NO and iNOS by 4-hydroxybenzyl alcohol (4-HBA), one of the active constituent of GE in LPS-stimulated BV-2 cells suggest that 4-HBA might be the bioactive candidate. CONCLUSION GE extract and its active constituent 4-HBA could be further exploited to mitigate microglial activation and may be developed as a new therapeutic remedy in treating various neuroinflammatory diseases.

Anti-inflammatory and anti-allergic effects of Agrimonia pilosa Ledeb extract on murine cell lines and OVA-induced airway inflammation

ETHNOPHARMACOLOGICAL EVIDENCE Agrimonia pilosa Ledeb (Rosaceae, AP) has long been used as a traditional medicine in Korea and other Asian countries to treat various diseases. AIM OF THE STUDY In the present study, the anti-inflammatory and anti-allergic effects of AP extract in in vitro cell lines and in vivo mouse model of inflammation and the molecular mechanisms involved were reported. MATERIALS AND METHODS Using Raw 264.7 murine macrophages the effects of methanol extract of AP in lipopolysaccharide (LPS)-induced production of inflammatory mediators were measured. Further IgE-DNP-induced interleukin (IL)-4 production and degranulation in RBL-2H3 rat basophilic cell lines was also estimated. To investigate the anti-asthmatic effect of AP in vivo, airway inflammation in ovalbumin (OVA)-induced mouse model was used. RESULTS AP attenuated the production of inflammatory mediators such as NO, PGE(2) and pro-inflammatory cytokines in LPS-induced Raw 264.7 cells. Further, AP inhibited IL-4 production and degranulation in IgE-DNP-induced RBL-2H3 cells. Furthermore, AP attenuated the infiltration of immune cells into lung, cytokines production in broncho-alveolar lavage fluid (BALF) and airway-hyperresponsiveness (AHR) on OVA-induced mouse model of inflammation. CONCLUSION Our results showed that AP attenuated the activation of macrophages, basophils, and inhibited the OVA-induced airway inflammation. The molecular mechanisms leading to APs potent anti-inflammatory and anti-allergic effects might be through regulation of TRIF-dependent and Syk-PLCγ/AKT signaling pathways, suggesting that AP may provide a valuable therapeutic strategy in treating various inflammatory diseases including asthma.