Leung Yiu Fai Ivan. / "July 2000." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2000. / Includes bibliographical references (p. 83-101). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.

FundaÃÃo Cearense de Apoio ao Desenvolvimento Cientifico e TecnolÃgico / O acidente vascular cerebral (AVE) à uma das principais causa de mortalidade no Brasil, acometendo cerca de 200.000 indivÃduos anualmente. A fisiopatologia do AVE isquÃmico envolve uma complexa cascata de eventos como a inflamaÃÃo e o estresse oxidativo que podem à morte neuronal e dÃficits cognitivos. O α-bisabolol à um Ãlcool sesquiterpeno, monocÃclico, que ocorre na natureza e à encontrado como constituinte majoritÃrio do Ãleo essencial sintÃtico da Matricaria chamomilla, que possui atividade antiinflamatÃria, antioxidante e anti-apoptÃtica jà descritas. Para avaliar o efeito neuroprotetor deste composto em camundongos submetidos à oclusÃo permanente da artÃria cerebral media (pMCAO), os animais foram prà e pÃs tratados com α-bisabolol nas doses de 50, 100 e 200 mg/kg, v.o, durante 24, 48, 72, 96 ou 120 horas apÃs a isquemia. Os animais foram avaliados 24h apÃs a isquÃmia para verificar a Ãrea de lesÃo isquÃmica, avaliaÃÃo neurolÃgica e atividade da mieloperoxidase. 72 horas apÃs a pMCAO, os testes de atividade locomotora, memÃria de trabalho e memÃria aversiva recente foram realizados. 96 horas apÃs a pMCAO foi realizado o teste do reconhecimento de objecto, e os animais foram eutanasiados para a realizaÃÃo da Imunohistoquimica para TNF-α, iNOS e GFAP e anÃlise histologica para Cresil violeta e Fluoro Jade C. Finalmente, 120h apÃs a isquemia, avaliou-se a memÃria espacial. O α-bisabolol reduziu significativamente a lesÃo isquemica e o dÃficit neurolÃgico e normalizou a atividade locomotora. O α-bisabolol mostrou proteÃÃo contra os dÃficits nas memÃrias de trabalho, espacial, reconhecimento de objeto e aversiva. O α-bisabolol (200 mg/kg) preveniu significativamente o aumento da MPO e TNF-α no cÃrtex temporal e o aumento do iNOS tanto no cÃrtex temporal como no estriado. Tambem preveniu o aumento da astrogliose nessas Ãreas. O α-bisabolol (200 mg/kg) mostrou protecÃÃo contra a morte neuronal. Os resultados do presente estudo mostraram que o α-bisabolol possui atividade neuroprotetora provavelmente devido a sua aÃÃo antiinflamatÃria, mas outros mecanismos nÃo podem ser descartados. / Stroke is the leading cause of mortality in Brazil, affecting about 200,000 individuals annually. The pathophysiology of ischemic stroke involves a complex cascade of events such as inflammation and oxidative stress which will lead to neuronal death and cognitive deficits. The α-bisabolol is a sesquiterpene alcohol, natural, monocyclic, found as main constituents of the essential oil of Matricaria chamomilla, which has anti-inflammatory, antioxidant and anti-apoptotic already described. To evaluate the neuroprotective effects of this compound in mice underwent permanent occlusion of the middle cerebral artery (pMCAO), the animals were pre and post treated with α-bisabolol at doses of 50, 100 and 200 mg / kg, orally for 24, 48, 72, 96 or 120 hours after pMCAO. The animals were evaluated 24 hours after ischemia to verify the area of ischemic damage, and neurological evaluation and myeloperoxidase activity. Seventy-two hours after pMCAO, the locomotor activity tests, working memory and aversive recent memory were performed. Ninety six hours after the pMCAO was performed the object recognition test, and the animals were euthanized for carrying out the immunohistochemistry for TNF-α, iNOS and GFAP and for histology analyes Cresil violet and Fluoro Jade C. Finally, 120 h after pMCAO, the spatial memory was evaluated. The α-bisabolol reduced significantly ischemic damage and neurological deficit and normalized the locomotor activity. The α-bisabolol showed protection against the deficits in working, spatial, object recognition and aversive memories. The α-bisabolol (200 mg / kg) significantly prevented the increase of MPO and TNF-α in the temporal cortex and the increased of iNOS both in the temporal cortex and in striatum. Also prevented the increase in astrogliosis in there area. The α-bisabolol (200 mg / kg) showed protection against neuronal death. The results of this study showed that α-bisabolol has neuroprotective activity probably due to its anti-inflammatory action, but other mechanisms can not be discarded.

Perinatal hypoxia-ischaemia is a major cause of disability, including cerebral palsy, yet a neuroprotectant which fully protects the brain remains elusive. Following a hypoxic-ischaemic insult, striatal medium-spiny neurons and hippocampal CA1 neurons are vulnerable to a complex cascade of neurotoxic events. This cascade includes energy failure, a massive release of glutamate, the formation of free radicals and caspase activation. The overall aim of this thesis was to assess the efficacy of three potential neuroprotective strategies that target this cascade from different directions. Short-term, and where appropriate, long-term, neuroprotection was investigated. The first treatment strategy aimed to suppress the generation of free radicals through treatment with the potent free radical spin trap, N-tertbutyl-(2-sulphophenyl)-nitrone (S-PBN). The second compound tested was the caspase-3 inhibitor, minocycline. Finally, the third treatment strategy combined a series of S-PBN injections with 6 hours of moderate hypothermia immediately after hypoxia-ischaemia. Hypothermia is suggested to slow the rate of the neurotoxic cascade, thus potentially allowing other neuroprotective agents greater efficacy. Using an adaptation of the Rice et al. (1981) model, hypoxia-ischaemia was induced on postnatal day (PN) 8 in the right cerebral hemisphere. For the short-term studies, the rats were perfused at 14 days-of-age. The brains were dissected out and embedded in Technovit. Forty [mu]m serial sections were cut through the right striatum and hippocampus. The total number of medium-spiny neurons in the striatum and where appropriate, the total number of neurons in the hippocampal CA1 pyramidal layer, were stereologically determined using the optical disector/Cavalieri method. For the long-term study, fine motor control was assessed in half of the animals through the staircase test from 9-11 weeks-of-age. Neuroprotection was assessed in the remaining animals. All animals were sacrificed at 12 weeks-of-age. The total number of striatal medium-spiny neurons was stereologically determined in the non-behavioural animals as described above. A series of seven injections of S-PBN (100mg/kg) did not offer statistically significant neuroprotection to the striatum at one week after perinatal hypoxia-ischaemia. Similarly, a single injection of minocycline (45mg/kg) immediately after the insult did not offer significant neuroprotection to the striatum nor the CA1 region of the hippocampus at this early time-point. In contrast, when the series of S-PBN injections was combined with 6 hours of moderate hypothermia post-hypoxia-ischaemia, sterelogical analysis revealed significant neuroprotection of the striatal medium-spiny neurons to normal levels at one week after the injury. No significant neuroprotection was seen in the CA1 region of the same animals. To assess whether this impressive striatal neuroprotection was long-lasting and whether it represented functional rescue, the final experiment in this thesis investigated rat pups at 12 weeks-of-age after exposure to hypoxia-ischaemia at PN8. Treatment with S-PBN/hypothermia offered persistent neuroprotection of striatal medium-spiny neurons and preservation of fine motor skills compared to diluent-normothermia-treated controls. The long-term behavioural outcomes were compared with normal, uninjured controls and the total number of medium-spiny neurons was compared with normal numbers from the literature. These comparisons revealed that the histological and functional integrity of the striatum was rescued to normal levels. This is the first study to identify a treatment strategy that offers complete and long-lasting preservation of striatal neuronal numbers, by accurate and unbiased stereological methods, paired with persistent preservation of fine motor control following perinatal hypoxia-ischaemia.

Subsequent to an hypoxic-ischaemic (HI)-insult a multi-faceted complex cascade of events occurs that ultimately results in cellular and neurological impairments within cortical and sub-cortical central nervous system (CNS) regions. In the present studies a modified �Levine� rat-pup model of HI (left carotid artery ligation + 1 hour global hypoxia on post-natal day (PND) 26) was employed to assess the neuroprotective properties of clomethiazole (CMZ; a γ-aminobutyric acid (GABA)A receptor agonist). In this study, histological and electrophysiological paradigms were used to assess the long-term neuroprotective properties of CMZ (414mg/kg/day via mini-pumps). Key enzymes involved in inflammation, namely nitric oxide synthase (NOS) and arginase, were also examined to assess other potential CMZ mechanisms. Assessments were carried out 3- and 90-days post-HI, with extensive ipsilateral CNS lesions evident at a gross histological level, at both the early and long-term stages, with CMZ significantly decreasing the lesion size at 3- and 90-days (P<0.01; P<0.05). Evoked field potential analyses were used to assess hippocampal CA1 neuronal activity ex vivo. Electrophysiological measurements contralateral to the occlusion revealed impaired neuronal function following HI relative to short- and long-term controls (P<0.001, 3- and 14-days; P<0.01, 90-days), with CMZ providing near complete protection (P<0.001 at 3- and 14-days; P<0.01 at 90-days). Both inducible NOS (iNOS) and arginase activities were significantly increased at 3-days (P<0.01), with arginase activity remaining elevated at 90-days post-HI (P<0.05) ipsilaterally. CMZ suppressed the HI-induced increase in iNOS and arginase activities (P<0.001; P<0.05). These data provide evidence of long-term functional neuroprotection afforded by CMZ in a model of HI-induced neurodegeneration. In addition, under conditions of HI, functional deficits were not restricted to the ipsilateral hemisphere and were due, at least in part, to changes in the activity of NOS and arginase. Underlying mitochondrial dysfunction is eminently present in many neuropathological conditions. The full extent of mitochondrial dysfunction in cortical, hippocampal and cerebellar tissues was assessed following HI. Assessment of mitochondrial FAD-linked respiration at both 1- and 3-days post-HI revealed a significant decrease in activity from ipsilateral cortical and hippocampal regions (P<0.001). In addition, significant changes in respiratory function were also evident in contralateral regions and cerebellum, 3-days post-HI (P<0.05). Assessment of the mitochondrial electron transport chain (complexes I-V) and mitochondrial markers of integrity (citrate synthase) and oxidative stress (aconitase) confirmed ipsilateral mitochondrial impairment following HI. Complexes I, II-III, V and citrate synthase were also impaired, in contralateral regions and cerebellum, 3-days post-HI. CMZ treatment provided significant protection to all mitochondrial aspects of neuronal tissue assessed. This study provides evidence of the full extent of mitochondrial damage following an HI-insult and may contribute, in part, to the impairment seen contralaterally. In addition, protection afforded by CMZ extends to preservation of mitochondrial function and integrity. Cerebral ischaemia-induced angiogenesis has been shown within and around infarcted regions and may contribute to a more favourable neurological outcome. The level of angiogenesis was examined using platelet endothelial cell adhesion molecule-1 (PECAM-1 / CD31). CD31 immunolabelling 7-days post-HI revealed a significant increase in angiogenesis compared with non-intervention controls (P<0.001). Treatment with CMZ decreased the level of angiogenesis compared to HI + saline (P<0.001) back to non-intervention control levels. Conversely, N[omega]-nitro-L-arginine methyl ester (L-NAME) treatment (5mg/kg/day) exacerbated the ischaemic lesion (P<0.001) and resulted in a marked decrease in angiogenesis compared to non-intervention controls (P<0.001). The extent of cerebral infarction in these studies is dependent on the level of NOS activity with CMZ increasing total NOS levels compared to HI + saline, while L-NAME halted the HI-induce increase in total NOS activity (P<0.001). These results show for the first time, that angiogenesis may be used as an assessment of neurodegeneration / neuroprotection in pathologies of cerebral ischaemia and are directly correlated with changes in NOS activity. These studies have therefore shown that following HI, damage also occurs contralateral to the occlusion, and is not restricted to the ipsilateral hemisphere. In addition, the neuroprotective effects of CMZ have been shown to extend out to 90-days post-HI, whereby significant protection to CA1 neuronal activity was seen. These studies also provide in vivo evidence that CMZ may also afford neuroprotection via anti-inflammatory pathways, as evidenced by a decrease in iNOS and arginase activities. Furthermore, these studies have also show evidence that angiogenesis (CD31) can be used as a diagnostic tool to assess neuroprotection / neurodegeneration.

published_or_final_version / Chinese Medicine / Doctoral / Doctor of Philosophy

Retinal detachment (RD) is a leading cause of blindness and although final surgical reattachment rate has greatly improved, visual outcome in many macula-off detachments is disappointing, mainly because of photoreceptor cell death. We previously showed that both lutein and Lycium barbarum polysaccharides (LBP) are neuroprotective in a rodent model of ischemia/reperfusion injury. The objective of this study is to investigate lutein and LBP as possible pharmacological adjuncts to surgery. Lutein: Subretinal injections of 1.4% sodium hyaluronate were used to induce RD in Sprague-Dawley rats until their retinae were approximately 70% detached. Daily injections of corn oil (control group) or 0.5mg/kg lutein in corn oil (treatment group) were given intraperitoneally starting 4 hours after RD induction. Animals were euthanized 3 days and 30 days after RD and their retinae were analyzed for photoreceptor apoptosis and cell survival at the outer nuclear layer (ONL) using TUNEL staining and cell counting on retinal sections. Glial fibrillary acidic protein (GFAP) and rhodopsin (RHO) expression were evaluated with immunohistochemistry. Western blotting was done with antibodies against cleaved caspase-3, cleaved caspase-8 and cleaved caspase-9 to delineate lutein’s mechanism of action in the apoptotic cascade. To seek a possible therapeutic time window, the same set of experiment was repeated with treatment commencing 36 hours after RD. When lutein was given 4 hours after RD, there was significantly fewer TUNELpositive cells in ONL 3 days after RD when compared with the vehicle group. Cell counting showed that there were significantly more nuclei in ONL in lutein-treated retinae by day 30. Treatment groups also showed significantly reduced GFAP immunoreactivity and preserved RHO expression. At day 3 after RD, Western blotting showed reduced expression of cleaved caspase-3 and cleaved caspase-8 in the treatment group. No difference was found for cleaved caspase-9. When lutein was given 36 hours after RD similar results were observed. Our results suggest that lutein is a potent neuroprotective agent that can salvage photoreceptors in rats with RD, with a therapeutic window of at least 36 hours. The use of lutein in patients with RD may serve as an adjunct to surgery to improve visual outcomes. LBP: The same RD model was used for the LBP experiment. Phosphate buffered solution (PBS) or LBP in PBS was given orally through a gavage at 1mg/kg and 10mg/kg concentrations. For this experiment, animals were sacrificed 7 days after RD, and only cell counting of the ONL and TUNEL staining were performed. Both sets of results did not produce statistically significant changes with the use of LBP. Our preliminary data for the effect of LBP on retinal detachment shows no significant beneficial effect. / published_or_final_version / Medicine / Master / Master of Research in Medicine

It has been known that light input can affect the emotions of a person. The depressive syndrome Seasonal Affective Disorder (SAD) is an effective example of the power of light in changing the mood of a person. Patients with SAD have recurring depressive episodes that follow seasonal changes, which is due to the changing daylight hours. This phenomenon suggests that there would be receptors in the retina that would not simply be responsible for vision, but also for the regulation of non-visual signals such as emotion. In many animals, projections have been found from the retina to the dorsal raphe nucleus (DRN). This brain region is a serotonergic area and has been found to be involved in the occurrence of depression. As such, the cells in the retina which were found to have projections to the DRN have a high possibility to be involved in emotion regulation. Retinal Ganglion Cells (RGCs) are classified into many types. A specific type known as an alpha cell is suspected to be the DRN-projecting subtype. This study uses Lycium Barbarum Polysaccharide (LBP) as a treatment in protecting the large RGCs from corticosterone (CORT) -induced damage. The aim is to observe if LBP will provide neuroprotection to large sized RGCs damaged by 40mg/kg or 50mg/kg CORT, and hence if LBP can be further investigated as a possible anti-depressant drug. This study observed that although LBP did not reduce large cell deaths, it reduced cell atrophy of the RGCs under high dosage of CORT (50mg/kg). For the same number of cells counted, treatment groups with a high dose CORT injection found more cells over 300μm2 in area than cells under 300μm2. Also, it was found that the temporal quadrants were more sensitive to cell size change than the nasal quadrants, paving way for more in-depth research of the spatial sensitivity to CORT or to LBP. The findings in this study indicate that LBP does indeed have a neuroprotective effect on large RGCs, although this effect is limited and as of yet seems conditional, as this study ignores the effect of CORT and LBP on other large cell properties such as the dendritic field size and the amount of synapses. Further studies are needed to determine the mechanism of the neuroprotective effect of LBP and to determine the exact site of action LBP works on. / published_or_final_version / Anatomy / Master / Master of Medical Sciences

Subsequent to an hypoxic-ischaemic (HI)-insult a multi-faceted complex cascade of events occurs that ultimately results in cellular and neurological impairments within cortical and sub-cortical central nervous system (CNS) regions. In the present studies a modified �Levine� rat-pup model of HI (left carotid artery ligation + 1 hour global hypoxia on post-natal day (PND) 26) was employed to assess the neuroprotective properties of clomethiazole (CMZ; a γ-aminobutyric acid (GABA)A receptor agonist). In this study, histological and electrophysiological paradigms were used to assess the long-term neuroprotective properties of CMZ (414mg/kg/day via mini-pumps). Key enzymes involved in inflammation, namely nitric oxide synthase (NOS) and arginase, were also examined to assess other potential CMZ mechanisms. Assessments were carried out 3- and 90-days post-HI, with extensive ipsilateral CNS lesions evident at a gross histological level, at both the early and long-term stages, with CMZ significantly decreasing the lesion size at 3- and 90-days (P<0.01; P<0.05). Evoked field potential analyses were used to assess hippocampal CA1 neuronal activity ex vivo. Electrophysiological measurements contralateral to the occlusion revealed impaired neuronal function following HI relative to short- and long-term controls (P<0.001, 3- and 14-days; P<0.01, 90-days), with CMZ providing near complete protection (P<0.001 at 3- and 14-days; P<0.01 at 90-days). Both inducible NOS (iNOS) and arginase activities were significantly increased at 3-days (P<0.01), with arginase activity remaining elevated at 90-days post-HI (P<0.05) ipsilaterally. CMZ suppressed the HI-induced increase in iNOS and arginase activities (P<0.001; P<0.05). These data provide evidence of long-term functional neuroprotection afforded by CMZ in a model of HI-induced neurodegeneration. In addition, under conditions of HI, functional deficits were not restricted to the ipsilateral hemisphere and were due, at least in part, to changes in the activity of NOS and arginase. Underlying mitochondrial dysfunction is eminently present in many neuropathological conditions. The full extent of mitochondrial dysfunction in cortical, hippocampal and cerebellar tissues was assessed following HI. Assessment of mitochondrial FAD-linked respiration at both 1- and 3-days post-HI revealed a significant decrease in activity from ipsilateral cortical and hippocampal regions (P<0.001). In addition, significant changes in respiratory function were also evident in contralateral regions and cerebellum, 3-days post-HI (P<0.05). Assessment of the mitochondrial electron transport chain (complexes I-V) and mitochondrial markers of integrity (citrate synthase) and oxidative stress (aconitase) confirmed ipsilateral mitochondrial impairment following HI. Complexes I, II-III, V and citrate synthase were also impaired, in contralateral regions and cerebellum, 3-days post-HI. CMZ treatment provided significant protection to all mitochondrial aspects of neuronal tissue assessed. This study provides evidence of the full extent of mitochondrial damage following an HI-insult and may contribute, in part, to the impairment seen contralaterally. In addition, protection afforded by CMZ extends to preservation of mitochondrial function and integrity. Cerebral ischaemia-induced angiogenesis has been shown within and around infarcted regions and may contribute to a more favourable neurological outcome. The level of angiogenesis was examined using platelet endothelial cell adhesion molecule-1 (PECAM-1 / CD31). CD31 immunolabelling 7-days post-HI revealed a significant increase in angiogenesis compared with non-intervention controls (P<0.001). Treatment with CMZ decreased the level of angiogenesis compared to HI + saline (P<0.001) back to non-intervention control levels. Conversely, N[omega]-nitro-L-arginine methyl ester (L-NAME) treatment (5mg/kg/day) exacerbated the ischaemic lesion (P<0.001) and resulted in a marked decrease in angiogenesis compared to non-intervention controls (P<0.001). The extent of cerebral infarction in these studies is dependent on the level of NOS activity with CMZ increasing total NOS levels compared to HI + saline, while L-NAME halted the HI-induce increase in total NOS activity (P<0.001). These results show for the first time, that angiogenesis may be used as an assessment of neurodegeneration / neuroprotection in pathologies of cerebral ischaemia and are directly correlated with changes in NOS activity. These studies have therefore shown that following HI, damage also occurs contralateral to the occlusion, and is not restricted to the ipsilateral hemisphere. In addition, the neuroprotective effects of CMZ have been shown to extend out to 90-days post-HI, whereby significant protection to CA1 neuronal activity was seen. These studies also provide in vivo evidence that CMZ may also afford neuroprotection via anti-inflammatory pathways, as evidenced by a decrease in iNOS and arginase activities. Furthermore, these studies have also show evidence that angiogenesis (CD31) can be used as a diagnostic tool to assess neuroprotection / neurodegeneration.

Thesis (Ph. D.)--University of Washington, 1998. / Vita. Includes bibliographical references (leaves [121]-138).

Parkinson's disease (PD) is one of the most common neurodegenerative diseases characterized by cell death in the substantia nigra pars compacta (SNpc) and the appearance of aggregated α-synuclein (α-syn). Autophagosomes accumulation and lysosomal reduction were discovered in PD patients' brain, which indicated the deficiency of autophagy in the progress of PD. TFEB (transcription factor EB) is a member of basic helix -loop-helix-leucine-zipper transcription factors (MiT family) and is a key master monitor for autophagy and lysosome biogenesis. Overexpression of TFEB is able to rescue the dopaminergic neurons (DAs) loss and α-syn aggregated in α-syn transgenic mice model and MPTP PD model. Hence, in recent years, many researchers have considered TFEB as a new therapeutic target for PD In this study, we discovered a novel TFEB activator named E4 by screening synthesized curcumin analogs. We have found that E4 strongly promoted TFEB nuclear translocation and induced autophagy in different cell lines. TFEB is essential for E4-induced autophagy flux. We also demonstrated that the underlying mechanism of E4 activate TFEB is mainly through inhibiting mTORC1 activity. We constitutively activate mTOR by knockdown TSC2 abrogated the increase of LC3-II and decrease of p-TFEB. We further estimated the protective effects of E4 in overexpressed α-syn model and neurotoxins induced cytotoxicity model. Treated with E4 for 48h in N2a transfected with A53T α-synuclein cells dose-dependently reduce the α-synuclein level. At the same time, we established the MPP+ model in PC12 cells which is pre-treated cell with E4 for 6 hours and then co-treated cells with MPP+ for 48 hours. The cell viability results showed that E4 significantly protect PC12 cells against MPP+ cytotoxicity dose-dependently. E4 had shown good neuroprotective effects in PD in vitro models while poor water solubility and low brain permeability restricted its application in PD animal models. Hence, assembling E4 molecules into self-carried nanoparticles (NanoE4) addressed the issue of poor water solubility and intranasal administration solved the problem of low permeability. In order to track NanoE4 release in vitro and in vivo, we further investigated the absorption and emission of NanoE4. However, the absorption fluorescence results showed that NanoE4 exhibits the strong aggregation-caused quenching effect (ACQ) due to π-π stacking of the planar molecule within the NPs. NanoE4 have much weak emission compared with E4 molecules. Therefore, we fabricated E4-TPAAQ NPs by co-reprecipitating E4 molecules with the reported fluorescent organic compound TPAAQ (2,6-Bis[4-(diphenylamino) phenyl] anthraquinone). Next, we developed an intranasal drug delivery system in our lab. After intranasal co-drop nanodrug E4-TPAAQ NPs for 24 hours, we observed strong fluorescence distributed in the brain which indicated that deliver nanoparticles into the brain successfully through nasal-brain system. Therefore, we examined the protective effect of NanoE4 in MPTP-induced PD mice model. In MPTP models, we found autophagy dysfunction, motor function decrease and increase of α-synuclein as reported previously. Treatment with NanoE4 rescued the motor dysfunction induced by MPTP. NanoE4 also increase TH level in the striatum part of midbrain. NanoE4 treatment also decreased the α-synuclein protein aggregate in both SNpc and striatum. Overall, these results demonstrate the neuroprotection NanoE4 against PD. Collectively, our findings 1) discovered a novel TFEB activator E4 that inhibited the mTOR pathway 2) indicated in vitro and in vivo experimental evidence for TFEB activator as the anti-PD drug candidate 3) provide a novel drug develop and delivery system for potential PD that limited by water solubility and BBB (blood-brain barrier) obstacle