Lipid droplets (LDs) are dynamic organelles implicated in Parkinson's disease (PD) pathology, yet their polarity dynamics and therapeutic relevance remain poorly understood. Herein, we rationally designed a polarity-responsive alkenyl indole-based fluorophore, PD3, through systematic molecular engineering to enable real-time tracking of LD polarity in PD models. By modulating intramolecular charge transfer (ICT) effects via electron-donating/withdrawing substitutions, PD3 exhibited exceptional polarity sensitivity, with a dramatic fluorescence enhancement in low-polarity environments and negligible viscosity interference. Density functional theory calculations confirmed that the polarity response originated from ICT-driven solvatochromism and solvent-dependent internal conversion rates. In rotenone-induced PD cellular models, PD3 revealed significant LD accumulation (4.2-fold intensity increase, 74% higher LD density, and 46% enlarged diameter) and organelle crosstalk, particularly with the endoplasmic reticulum (ER) and mitochondria. Furthermore, PD3-enabled evaluation of PD drugs demonstrated that amantadine (AMA) and pramipexole (PPX) restored LD homeostasis, linking their therapeutic effects to LD modulation. Ex vivo studies in methyl-phenyl-tetrahydropyridine (MPTP)-induced PD mice confirmed LD accumulation in brain tissues, correlating with motor deficits and neuronal loss in the substantia nigra. This work establishes PD3 as a powerful tool for deciphering LD-associated PD pathology and advancing LD-targeted therapeutic strategies.

ABSTRACT Circadian rhythm disruption has been increasingly implicated in neurodegenerative diseases, including Parkinson’s disease (PD). While motor impairments in PD are well-established, the impact of chronic circadian misalignment on both motor and cognitive dysfunctions remains underexplored. This study investigates the effect of prolonged exposure to different lighting conditions on MPTP-induced motor and cognitive deficits in a rat model. A total of 100 adult male Wistar rats were divided into five groups with different light-dark cycle conditions: standard 12-h light/12-h dark (12 L:12D), constant light (24 L), constant darkness (24D), 20-h light/4-h dark (20 L:4D), and 4-h light/20-h dark (20D:4 L). After 60 d of exposure to these conditions, each group was further divided into Parkinsonian (MPTP-treated) and non-Parkinsonian (saline-treated) subgroups. MPTP hydrochloride (25 µg/kg) was administered intranasally to induce Parkinsonism. Behavioral assessments, including the beam balance test, open field test, Y-maze test, and shuttle box test, were conducted to evaluate motor coordination and cognitive function. Immunohistochemical analyses of α-synuclein (α-syn) accumulation in the substantia nigra (SNc) and tyrosine hydroxylase (TH) expression in the suprachiasmatic nucleus (SCN) were performed. Data were analyzed using t test, one-way ANOVA and Tukey’s post hoc test. MPTP administration significantly impaired motor function, as evidenced by increased crossing time in the beam balance test (p < 0.001) and reduced locomotor activity in the open field test (p < 0.01). Prolonged exposure to different lighting conditions further exacerbated these deficits, particularly in the 24 L and 20 L:4D groups (p < 0.0001). Cognitive assessments revealed that MPTP-treated rats exhibited marked deficits in working memory (Y-maze, p < 0.05) and associative learning (shuttle box, p < 0.01), with the most severe impairments observed in groups subjected to extreme light-dark alterations. Immunohistochemical analyses demonstrated significantly increased α-syn accumulation in the SNc (p < 0.0001) and decreased TH expression in the SCN (p < 0.001), particularly in MPTP-treated rats under photoperiod disruption. Chronic photoperiod disruption exacerbates MPTP-induced motor dysfunction, cognitive impairment, and neurodegeneration. Targeting circadian regulation may offer a potential therapeutic approach for mitigating neurodegenerative progression in PD.

Coriandrum sativum Linn. (family: Umbelliferae; C. sativum), is a potential herb widely used as a spice and traditional medicine. In the present work, the effects of C. sativum fruit extract (CSE), against lipopolysaccharide (LPS)-stimulated BV-2 microglia-mediated neuroinflammation in vitro and 1-methyl-4 phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP)-induced Parkinson's disease (PD) animal model in vivo was investigated. LPS-stimulated increase in nitric oxide (NO), inducible NO synthase, cyclooxygenase-2, interleukin-6 and tumor necrosis factor-alpha were significantly (p < 0.05 ~ p < 0.001) inhibited by CSE (25, 50 and 100 μg/mL) in BV-2 microglial cells. Further, CSE inhibited the LPS-induced nuclear factor of kappa-beta activation and IκB-α phosphorylation in BV-2 microglia. In vivo studies, CSE (100, 200 and 300 mg/kg) ameliorated the MPTP (25 mg/kg, i.p.)-induced changes in locomotor, cognitive and behavior functions evaluated by rotarod, passive avoidance and open field test significantly (p < 0.05 ~ p < 0.001). The MPTP-induced changes in brain oxidative enzyme levels such as superoxide dismutase, catalase, and lipid peroxidation were significantly (p < 0.01 and p < 0.001 at 200 and 300 mg/kg, respectively) restored with CSE treatment. High-performance thin-layer chromatography fingerprinting analysis of CSE exhibited several distinctive peaks with quercetin and kaempferol-3O-glucoside as identifiable compounds. In conclusion, our study indicated that CSE attenuated neuroinflammatory processes in LPS-stimulated microglia in vitro and restored the MPTP-induced behavioral deficits and brain oxidative enzyme status in vivo proving its therapeutic potential in the treatment of neuroinflammatory and oxidative stress-mediated neurodegeneration seen in PD.

β-Caryophyllene exerts protective antioxidant effects through the activation of NQO1 in the MPTP model of Parkinson’s disease

Objective To investigate the protective effects of stilbene glycoside(2, 3, 5, 4'-tetrahydroxy-stilbene-2-O-β-D-glucoside, TSG)on 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine(MPTP)-induced mice model of Parkinson's disease(PD). Methods Mice were randomly divided into the blank control group, the negative control group, the TSG high-dose group, the TSG low-dose group and the positive drug group(n=20 each). Mice were weighted daily to observe the changes of body weight, and mice motor and behavior function were tested by open field test.Level changes of α/β synuclein in brain cortex, cerebellum, midbrain, and hippocampal were detected by Western blot. Results Compared with the blank control group, the negative control group showed that the body weight was decreased(P<0.05). Compared with the negative control group, the body weight was increased in the TSG high-and low-dose groups and the positive drug group(P<0.05). The spontaneous behavior was impaired in the negative control group.Compared with the blank control group, the negative control group showed that the open field test showed traveled distance over a 10-min period was significantly shortened at 1st, 7th, 28th days after testing(all P<0.05). The trajectory of motor axons indicated that mice in the negative control group showed dyskinesia, but the groups of positive drug and high-and low-dose of TSG could reverse this dyskinesia.Compared with the blank control group, brain α/β synuclein protein levels were increased in the negative control group, and decreased in positive drug and TSG high-and low-dose groups(P<0.05). Conclusions Stilbene glycosides exert neuroprotective effects in MPTP-induced mice model of PD. Key words: Parkinson disease; Stilbenes; 1-methy-4-phenyl-1, 2, 3, 6-tetrahydropyridine

BackgroundParkinson's disease is a progressive degenerative nervous system disease. Recent studies have shown that secondary changes in the GABA system play directly affect the pathogenesis of PD. There is still much debate about GABA concentrations because currently, GABA concentrations in the brain tissue are obtained indirectly by measuring its concentration in the blood and cerebrospinal fluid. These results are unreliable. Magnetic resonance spectroscopy (MRS) is the only noninvasive method for evaluating the concentration of metabolites in living brain tissue and has been widely applied in research and clinical practice. In addition, combining MEGA-PRESS technology with LCModel software for quantitative GABA measurements is largely recognized. At present, the PD monkeys model in primates has been increasingly proficient. Primates are more similar to humans in terms of brain structure and function than other animals. However, 3.0 T MRS studies involving the PD monkey model to measure metabolites in living subjects with PD are still rare. The study was performed at 3.0 T MRI with control monkeys and PD monkeys that were injected methyl-phenyl-tetrahydropyridine (MPTP) in one side of common carotid artery before and 3 months after successful model establishment to measure GABA concentrations in the bilateral striatum. Behavioral observations were performed for all animals, and the behavioral score was recorded. After 3 months, the GABA concentration in the bilateral striatum was measured in both groups by high-performance liquid chromatography (HPLC). The data obtained from magnetic resonance spectroscopy (MRS) were compared with the actual measured GABA concentrations in tissues isolated from the corresponding regions, and their correlations with the behavior score were analyzed. The research objectives are to investigate the changes of γ-aminobutyric acid (GABA) concentration in the bilateral striatum of monkeys with Parkinson's disease (PD) and the value of quantitatively measuring its concentration by noninvasive 3.0 T spectroscopy.Results(1) The MRS results showed that the GABA concentration in the injured side of the striatum of the PD monkeys was higher than in the contralateral side, but the difference was not statistically significant (P = 0.154). Compared with that the blank control group, the GABA concentration in the striatum of the PD monkeys increased, but there was no difference between the groups (P = 0.381; P = 0.425). (2) The GABA concentration that determined from the isolated specimens by HPLC in the injured side of the striatum of the PD monkeys was significantly higher than that in the contralateral side (P < 0.01). Compared with the blank control group, the PD monkeys had higher GABA concentrations in both sides of the striatum, and there was a significant difference in the lesion side (P = 0.004), while there was a non-significant difference in the contralateral side (P = 0.475). (3) The mean GABA concentration in the injured striatum of PD monkeys determined by MRS was not significantly correlated with the behavioral score (r = 0.146, P = 0.688). The mean GABA concentration in the injured striatum determined from the isolated specimens was positively correlated with the behavioral score in the same period (r = 0.444, P = 0.038).ConclusionThe GABA concentration in the injured striatum of PD monkeys is increased and positively correlated with behavioral changes. Validity of noninvasive 3.0 T MRS to detect PD neurotransmitter changes is limited.

While progressive dopaminergic neurodegeneration is responsible for the cardinal motor defects in Parkinson's disease (PD), new diagnostics and therapeutic targets are necessary to effectively address this major global health burden. We evaluated whether the adhesion G protein-coupled receptor B1 (ADGRB1, formerly BAI1, brain-specific angiogenesis inhibitor 1) might contribute to dopaminergic neuronal loss. We used bioinformatic analyses, as well as in vitro and in vivo PD models. We report in this study that ADGRB1 is decreased in PD and that the ADGRB1 level is specifically decreased in dopaminergic neurons in the substantia nigra of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-intoxicated mice. In primary mouse mesencephalic neurons and human neuroblastoma cell lines, 1-methyl-4-phenylpyridinium (MPP+), a toxic metabolite of MPTP, suppressed the expression of ADGRB1. Moreover, we applied a network generation tool, Ingenuity Pathway Analysis®, with the transcriptomics dataset to extend the upstream regulatory pathway of ADGRB1 expression. AMP-activated protein kinase (AMPK) was predicted as a regulator, and consequently, 5-aminoimidazole-4-carboxamide ribonucleotide, a specific activator of AMPK, reduced the ADGRB1 protein level. Finally, ADGRB1 overexpression decreased nuclear condensation induced by MPP+ treatment. Taken together, we observed that decreased ADGRB1 by activation of AMPK induced neuronal cell death in MPTP/MPP+-mediated PD models, suggesting that ADGRB1 might potentially play a survival role in the neurodegenerative pathway of PD. These data offer new insights into dopaminergic cell death with therapeutic implications for neurodegenerative disorders.

Adult male mice C57BL/6 (n = 105) were divided into five groups. The first group served as a control. In the 2nd–5th groups, the animals were treated subcutaneously with 40 mg/kg of proneurotoxin MPTP (methylphenyltetrahydropyridine), which forms a state similar to the initial stage of Parkinson’s disease over a 2-week period. Mice of groups 3–5 daily received an additive along with their food: one of three extracts of the biomass of the litter beetle Alphitobius diaperinus. In 2 weeks, all animals were tested for motor disorders in the vertical bar test; they were then euthanized and histochemical analysis of the dopamine-containing brain regions was performed. In addition, the same extracts were tested for counteraction to MPP+ toxin in cultured neuroblastoma cells. It was found that the primary aqueous and, especially, secondary water–methanol extracts had a powerful protective effect against the neurotoxic effect judging by the results of both the behavioral test and morphological control. Arginine was found at substantial concentrations in both effective extracts. An in vitro study confirmed the protective effect of the primary aqueous extract.

Five groups of C57Bl/6JSto mice (groups 1–5) were tested on a vertical pole two weeks after s.c. administration (groups 2–5) of the proneurotoxin methylphenyltetrahydropyridine (MPTP) at a dose of 40 mg per kg, which at this time induces sensorimotor impairments similar to the early stage of Parkinson’s disease. Mice of group 1 served as controls. One week before MPTP administration and throughout the whole of the post-treatment period (two weeks), animals of groups 3–5 received a feed supplement (8 g/kg of feed) consisting of one of three extracts of biomass homogenate of the “remedy” beetle Alphitobius diaperinus. Extracts were: aqueous (No. 1), aqueous-methanolic (No. 2), and aqueous-methanolic after solid-phase extraction (No. 3). Extracts were immobilized on a plant-based carrier. Animals of groups 1 and 2 were kept on a standard diet without supplementation. The severe sensorimotor impairments seen in mice of group 2 (toxin) did not occur in either group 1 (controls) or groups 3–5, where animals received dietary supplementation with “antidote” consisting of darkling beetle biomass extract. The most effective was preparation No. 3.

MPTP (1-metyl 4-phenyl 1,2,3,6-tetrahydro-piridin) treatment is a validated toxic model to produce experimental parkinsonism. Chronic exposure to little amounts of MPTP has been reported to reproduce neuropathological features characterizing dopaminergic neurons in Parkinson disease (1). Recent studies by our group have found that MPTP, besides dopaminergic neurons, affects also spinal motor neurons (2), thus reproducing also the neuropathology of spinal cord in Parkinson’s disease. The aim of the present study is to investigate, through immune-histochemistry, the involvement of spinal motor neurons after chronic administration of MPTP. Chronic MPTP treatment is able to induce motor neurons loss in the lumbar spinal cord. Increased immune-reactivity of the autophagy protein MAP LC3β is detectable in motor neurons which indicates the activation of autophagy. Immune-staining for alpha-synuclein, the hallmark protein for Parkinson’s disease, was altered in spinal motor neurons of MPTP treated mice, as detected by two novel homemade monoclonal antibodies. Furthermore, immunohistochemical alterations have been detected in the Renshaw cells and in tyrosine-hydroxylase-containing axons in line with the alterations of noradrenergic projections in Parkinsons’s disease (3). Our results suggest that spinal cord may contribute to motor symptoms occurring in Parkinson’s disease.

Objective To investigate the effects of rapamycin which is an autophagy inducer and 3- adenine (3-MA) which is an autophagy inhibitor on motor behavior and autophagy related protein LC3 in C57BL/6 mice of Parkinson's disease(PD). Methods 40 C57BL/6 male mice were randomly divided into control group and experimental groups which consist of MPTP model group, rapamycin group and 3-MA group, with 10 in each group. Mice in experimental groups received intraperitoneal injection with MPTP to establish PD models, and mice in control group received intraperitoneal injection with the same amount of saline solution for 1 week. Mice in rapamycin group received intraperitoneal injection with rapamycin and mice in 3-MA group received intraperitoneal injection with 3-MA at the same time when MPTP was injected.Animal behavior detections were carried out on the 1th, 7th and 14th day after the last injection.After the last injection, mice were sacrificed and sections of midbrain nigra were gained for the detection of expression of autophagy specificity protein LC3 by Western Blot. Results Compared with MPTP model group, rapamycin could improve the general condition and behavioral manifestation of mice in pole test((14.89±1.14)s vs (16.24±1.39)s, P<0.05; (15.18±1.36)s vs(17.93±1.11s), P<0.01), traction test((1.7±0.5) vs (1.2±0.4), P<0.05; (1.5±0.5)vs (1.1±0.3), P<0.05)and open field test which contained total activity distance((5 875.3±1148.9)cm vs (5 061.5±773.1)cm, P<0.05; (5 753.2±1 106.7)cm vs (4 669.3±969.1)cm, P<0.01)and average speed((19.29±2.35)cm/s vs (16.47±3.01)cm/s, P<0.05; (18.71±2.71)cm/s vs (15.80±2.50)cm/s, P<0.01), while 3-MA aggravated behavioral deficits of mice in pole test(19.92±1.61s vs 17.93±1.11s, P<0.05), and both total activity distance((3 879.7±575.0)cm vs (4 669.3±969.1)cm, P<0.05)and average speed((13.34±1.87)cm/s vs(15.80±2.50)cm/s, P<0.05)in open field test were decreased.The results of Western Blot confirmed that rapamycin could increase the expression of LC3-Ⅱ, however 3-MA could reduce the expression of LC3-Ⅱ. Conclusion This study confirmed that rapamycin could alleviate behavioral symptoms of PD model mice and increase the level of LC3 in midbrain nigra, while 3-MA could exacerbate behavioral symptoms in PD model mice and decrease the level of LC3 in midbrain nigra.Thus it can be speculated that drugs which can regulate autophagy may be potential treatment protocols for PD. Key words: Rapamycin; 3-MA; Parkinson’s disease; LC3; Autophagy

Objective To investigate the anxious behavior in acute parkinson's mice that were induced by 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP) injection. Methods Twenty mice were randomly divided into the control group (n=10) and model group(n=10); The model group was induced by injecting MPTP dosage, and the control group was induced by the same dose of saline. The anxious behaviors in mice were tested by the elevated plus-maze test and the light/dark box. Results The model group mice spent a longer time than the control group in the dark box (P<0.05=. The open arm entry (OE), open arm time (OT) and OE% of model group was significantly less than that in control group in the elevated plus-maze test (P<0.01=, the OT% was significantly less than control group(P<0.05=. Conclusions Anxiety symptoms appeared in the model group of early arkinson disease (PD) mice. Key words: 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine/AE; Parkinson disease/CO/ET; Anxiety/CO

BACKGROUND Ischemia damages the electron transport chain (ETC), including at complex I. The transient, reversible blockade of complex I during early reperfusion decreases cardiac injury. Our studies in isolated mitochondria showed that high dose Metformin (MET) partially inhibits complex I in mitochondria that were damaged by ischemia. In contrast to previous chronic pretreatment studies, we asked if partial blockade of complex I only at the onset of reperfusion using high dose MET (2 mM), without pretreatment, would decrease cardiac injury. Opening of the mitochondrial permeability transition pore (MPTP) is a key effector of ETC‐mediated injury during reperfusion. METHODS Buffer‐perfused mouse hearts (C57BL/6) were subjected to 25 min ischemia and 30 min (mitochondrial isolation) or 60 min (infarct size) reperfusion with or without MET (2 mM) treatment for the first 5 min. of reperfusion only. Infarct size was used to evaluate myocardial injury. Opening of MPTP was evaluated by calcium retention capacity (CRC, nmol Ca 2+ /mg) in isolated mitochondria. Oxidative phosphorylation (OXPHOS) was measured. RESULTS High dose MET treatment at reperfusion decreased infarct size and improved contractile function compared to untreated hearts. MET decreased susceptibility to MPTP opening (improved CRC). OXPHOS rate was not improved, since much of the damage occurs during ischemia, before MET treatment during reperfusion. Thus, MET treatment decreases MPTP opening during reperfusion. CONCLUSION Administration of MET only during reperfusion protects hearts during ischemia‐reperfusion by partial blockade of complex I with decreased MPTP opening. Regional treatment of myocardium with high dose MET in the cardiac cath lab at the time of stent deployment for acute myocardial infarction is a clinically translatable approach to cardiac protection via modulation of the ischemia‐damaged ETC during early reperfusion. Support or Funding Information This work was supported by the Office of Research and Development, Medical Research Service, Department of Veterans Affairs, a CCTR grant from Virginia Commonwealth University, a Scientist Development Grant from the American Heart Association and the Pauley Heart Center of Virginia Commonwealth University. Isolated hearts Infarct size LVDP LVEDP IR 37±4 7±1 47±4 MET + IR 15±3 18±8 38±2 Isolated mito OXPHOS (nAO/min/mg) CRC substrates Glu + Mal Succ + Rot TC (n=5) 403±39 530±34 723±37 IR (n=6) 254±30 396±44 350±12 MET+IR (n=G) 216±25 367±18 467±22 Mean ± SEM; LVDP‐developed pressure mmHg; LVEDP‐end of diastolic pressure; Complex I substrates are glutamate (Glu) + malate (Mal); complex II substrates are succinate (Succ) + rotenone (ROT); p&lt;0.05 vs. ischemia‐reperfusion (IR); p&lt;0.05 vs. TC (time control).

Objective To study the neuroprotective role of TFP5 in a MPTP-induced mouse model of Parkinson's disease (PD). Methods C57BL/6 mice were used as experimental animals. Briefly, 5 consecutive days of intraperitoneal injection of 25 mg/Kg 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP) was applied to induce mouse PD model. The mice were randomized into 5 groups including control group, model group, scrambled TFP5 peptide (Scb) group, TFP5 group and roscovitine group. On the 7th day after the first injection of MPTP, behavior tests were performed, and then western blot method was employed to detect the expression of p25 and phosphorylated MEF2D in substantia nigra.Tyrosine hydroxylase (TH) immunohistochemical staining was performed to observe the apoptosis of dopaminergic neurons in substantia nigra pars compacta (SNpc) 28 days after the first injection of MPTP. Results MPTP increased the expression of p25 (0.48±0.10 vs 0.26±0.02, P<0.05) and phosphorylated MEF2D (0.81±0.10 vs 0.22±0.02, P<0.05) in substantia nigra, but decreased the number of dopaminergic neurons in SNpc (348.67±24.40 vs 463.29±19.61, P<0.05), resulting in motor impairment in the model mice (P<0.05). Intraperitoneal injection of 30mg/Kg of TFP5 for 3 days effectively reduced the excessive phosphorylation of MEF2D (0.25±0.12 vs 0.81±0.10, P<0.05) in substantia nigra, rescued dopaminergic neuron reduction of SNpc (422.92±8.41 vs 348.67±24.40, P<0.05), and improved the motor ability of the model mice (P<0.05). Roscovitine exerted almost same neuroprotective role as TFP5, while Scb had no protective effect. Conclusion TFP5 can rescue MPTP-induced damage of dopaminergic neurons in substantia nigra, and thus improve motor impairment of model mice, which may be mediated by the inhibition of Cdk5/p25 activity. Key words: Parkinson's disease; Cyclin-dependent kinase 5; TFP5; Dopaminergic neurons

Objective To investigate the effects of transcranial ultrasound stimulation (TUS) on the motor functioning and anti-oxidative capacity of mice with Parkinson's disease (PD). Methods Thirty-two inbred C57BL male mice were randomized into a normal control group, a model group, a sham TUS group and a TUS group (n=8 for each group) according to a random number table. A PD model was induced in the mice of the model, sham TUS and TUS groups by injecting 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP) at 20 mg/kg intraperitoneally, while those in the normal control group were given saline. Low intensity (1 W) focused ultrasound (LIFU) at a frequency of 0.5 MHz was then applied to stimulate the nigra region, except for the mice in the sham TUS group, which were treated with the same procedure but with no ultrasound output. A pole climbing test was carried out before, 2 weeks and 5 weeks after the injection of the MPTP. After 5 weeks the animals were sacrificed and the whole brain malondialdehyde (MDA) and glutathione peroxidase (GSH-Px) content were measured. Results No significant differences in in pole climbing scores were observed among the four groups before the MPTP injections. However, the average value decreased significantly to (4.30±1.19), (4.40±0.23) and (4.80±0.23) for the model, sham TUS and TUS groups respectively 2 weeks after the injection. It then rose to (5.12±0.83) and (5.51±1.21) for the first two groups 3 weeks later, but was still lower that before the injection. After 5 weeks the TUS group's average score was significantly higher than 3 weeks earlier and than that of the model group and the sham TUS group. Compared with the control group, the other groups' average scores were all lower 2 weeks after MPTP injection, and those of the model and the sham TUS groups remained so 5 weeks after the injection. Five weeks after the injection, the average MDA content of the model group (10.2±1.1 nmol/ml) and the sham TUS group (9.4±1.3 nmol/ml) were significantly higher than the normal control group (4.5±0.8 nmol/ml), as well as the TUS group (6.8±0.9 nmol/ml). However, GSH-Px enzyme activity in the model group (100±35.4 U/mgprot) and the sham US group (444±24.9 U/mgprot) was significantly lower than that in the normal control group (1262.5±53 U/mgprot), together with the TUS group (1047.3±77.8 U/mgprot). Conclusion TUS can improve motor function in PD, at least in mice. This may be due to its anti-oxidative capacity. Key words: Ultrasound; Parkinson's disease; Motor function; Antioxidants; Brain

Objective To investigate the neuroprotective effect of geniposide on 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP) induced Parkinson's disease (PD) model mice and possible mechanism. Methods A total of 48 male C57BL/6 mice were randomly divided into control, model, geniposide and MPTP + geniposide groups. The behaviors of C57BL/6 mice were assessed by using open field test, the tyrosine hydroxylase (TH) and Bcl-2 positive neurons in the midbrain substantia nigra of mice were detected by immunohistochemistry and the number of apoptosis neurons were observed with TdT-mediated dUTP-biotin nick end labeling (TUNEL). Results The number of mobile grid [(76.33 ± 8.59) times/5 min], standing [(19.58 ± 3.97) times/5 min], TH-positive neurons [(12.83 ± 2.32)/HPF] and Bcl-2-positive neurons [(10.83 ± 2.23)/HPF] in model group were significantly lower than those in the control group [(142.50 ± 11.65) times/5 min, (39.17 ± 4.75) times/5 min, (35.67 ± 1.75)/HPF, (20.67 ± 1.75)/HPF; P = 0.000, for all]. The apoptosis neurons in model group [(20.33 ± 2.58)/HPF] were significantly higher than that in control group [(3.83 ± 1.67) /HPF, P = 0.000). The number of mobile grid [(97.67 ± 13.15) times/5 min, P = 0.000], standing [(29.33 ± 2.90) times/5 min, P = 0.000], TH-positive neurons [(17.50 ± 2.07)/HPF, P = 0.002] and Bcl-2-positive neurons [(15.17 ± 2.79) /HPF, P = 0.003] in MPTP + geniposide group were significantly higher than those in model group. The number of apoptosis neurons [(14.67 ± 3.08) /HPF] in MPTP + geniposide group was significantly lower than that in model group ( P = 0.001). Conclusions Geniposide can protect dopaminergic neurons in MPTP-induced neurodegeneration and the mechanism may be associated with the inhibition of neuronal apoptosis. DOI: 10.3969/j.issn.1672-6731.2015.06.012

Insects have long attracted the attention of researchers as a source of biologically active sub� stances present in the tissues of different species of insects at various stages of their life cycle (1, 2). The analysis of the chemical composition of some species of invertebrates showed that the bee ( Apis mellifera), grasshoppers (family Acrididae), housecricket (Acheta domesticus), American cockroach (Periplan� eta americana), larval and adult mealworms (Tenebrio molitor), as well as larvae of the wax moth (Galleria mellonella), silkworm (Bombyx mori), blowfly (family Calliphoridae), black soldier fly (Hermetia illucens), etc., contain not only proteins and fats, but also chitin, minerals, amino acids, polyunsaturated fatty acids and vitamins, essential oils, and other biologically active sub� stances, including new antimicrobial peptides (3-9). In the scientific literature, there is scarce information about the possible medical use of extracts from the darkling beetle Ulomoides dermestoides from the family Tenebrionidae, which exhibit antiinflammatory and immunomodulating effect (10) and cytotoxicity with respect to A549 (lung adenocarcinoma) tumor cells (11). A species close to Ulomoides dermestoides is the beetle Alphitobius diaperinus, which is also used in folk medicine to treat various diseases, including one of the most socially important—Parkinson's disease. It was assumed that the Alphitobius diaperinus biomass homogenate may exhibit inhibitory activity with respect to MPTP (methylphenyl tetrahydropyri� dine)—a toxin that causes experimental parkinsonism in C57BL/6JSto mice (12, 13).

Although selective neurodegeneration of nigro-striatal dopaminergic neurons is widely accepted as a cause of Parkinson's disease (PD), the role of vascular components in the brain in PD pathology is not well understood. However, the neurodegeneration seen in PD is known to be associated with neuroinflammatory-like changes that can affect or be associated with brain vascular function. Thus, dysfunction of the capillary endothelial cell component of neurovascular units present in the brain may contribute to the damage to dopaminergic neurons that occurs in PD. An animal model of PD employing acute, sub-acute and chronic exposures of mice to methyl-phenyl-tetrahydropyridine (MPTP) was used to determine the extent to which brain vasculature may be damaged in PD. Fluoro-Turquoise gelatin labeling of microvessels and endothelial cells was used to determine the extent of vascular damage produced by MPTP. In addition, tyrosine hydroxylase (TH) and NeuN were employed to detect and quantify dopaminergic neuron damage in the striatum (CPu) and substantia nigra (SNc). Gliosis was evaluated through GFAP immunohistochemistry. MPTP treatment drastically reduced TH immunoreactive neurons in the SNc (20.68 ± 2.83 in acute; 22.98 ± 2.14 in sub-acute; 10.20 ± 2.24 in chronic vs 34.88 ± 2.91 in controls; p<0.001). Similarly, TH immunoreactive terminals were dramatically reduced in the CPu of MPTP treated mice. Additionally, all three MPTP exposures resulted in a decrease in the intensity, length, and number of vessels in both CPu and SNc. Degenerative vascular changes such as endothelial cell 'clusters' were also observed after MPTP suggesting that vasculature damage may be modifying the availability of nutrients and exposing blood cells and/or toxic substances to neurons and glia. In summary, vascular damage and degeneration could be an additional exacerbating factor in the progression of PD, and therapeutics that protect and insure vascular integrity may be novel treatments for PD.

Parkinson Disease (PD) is the prototype of synucleinopathies, a group of neurodegenerative disorders characterized by the accumulation of alpha-synuclein (α-syn). The involvement of spinal cord (SC) in PD is described in human autopsies, clinical reports and experimental studies [1]. The neurotoxin MPTP reproduces to a remarkable extent neuropathology of PD [2]. Furthermore, recent studies have shown that MPTP, besides dopaminergic neurons, affects also spinal Motor Neurons (MNs) [3]. To detect the effects of different MPTP treatments on the neuropathology in the SC at different ages and in different animal species. Following MPTP, neurotoxicity involves the whole thoraco-lumbar spinal cord. A model-dependent variation in the pattern of α-syn immunostaining has been detected. MNs, sympathetic pre-ganglionic neurons and calbindin immunoreactive (IR) cells of lamina VII are mostly affected, whereas TH positive fibers are spared. Alterations in α-syn positive fibers occur in the posterior laminae (I, II and III) and correlate with a significant loss of Met-Enk, SP and calbindin 28 kDa. A-syn accumulates in the affected neurons and in glial cells. Our results indicate that the SC represents a target of the parkinsonism-inducing neurotoxin MPTP. These data suggest that altered α-syn distribution associates with MPTP-induced neurotoxicity.

Objective To investigate the effect of repetitive transcranial magnetic stimulation (rTMS) on dopaminergic neurons in substantia nigra and nigrostriate fibers in the striatum in cases of Parkinson's disease (PD).Methods Forty male C57BL/6J mice were randomly divided into a control group,a PD model group,a sham-rTMS (srTMS) group and an rTMS group with 10 mice in each group.A PD model was established by subcutaneous injection of a solution of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP),and the mice were treated with rTMS or sham rTMS for 14 days.Tyrosine hydroxylase (TH) expression in the substantia and striatum were detected using an immunohistochemical technique,and the corrected optical densities (CODs) of TH in the striatum were analysed using an image analysis system.The Nissl bodies were detected by Nissl staining.The morphological disposition of nerve fibers in the striatum was detected using Warthin-Starry staining.Results The fraction of neurons expressing TH decreased significantly more in the substantia nigras of mice in the rTMS group than in the control group,the PD group and the srTMS group.In the control group the neural plasm of dopaminergic neurons was full of dark blue and granular Nissl bodies.Many Nissl bodies were lost in the PD and srTMS groups,and the remaining Nissl bodies were colored lightly.Few Nissl bodies were lost in the rTMS group.Positive TH reactions in the striatum were significantly decreased in the rTMS group compared to the control group.The average COD was also significantly lower.But positive TH reactions in the striata of the rTMS group mice were significantly greater than in the PD and srTMS groups,and the average COD was significantly higher.In the control group,the disposition of nerve fibers in the striatum was typically fasciculated,concentrated and ordered; in PD and srTMS groups,many fibers were lost,and the remaining nerve fibers were rare,ruptured and scattered ; in the rTMS group fewer nerve fibers were lost and the disposition of the remaining fibers was more fasciculated,concentrated and ordered than in the PD and srTMS groups.Conclusion rTMS may play a role in treating Parkinson's disease by protecting dopaminergic neurons as well as nigrostriate fibers and by improving the synthesis and transport of dopamine. Key words: Transcranial magnetic stimulation; Substantia nigra; Corpus striatum; Parkinson's disease