Adoptive therapy with amyloid-ß specific regulatory T cells alleviates Alzheimer's disease.

Rationale: Neuroinflammation is a primary feature of Alzheimer's disease (AD), for which an increasing number of drugs have been specifically developed. The present study aimed to define the therapeutic impact of a specific subpopulation of T cells that can suppress excessive inflammation in various immune and inflammatory disorders, namely, CD4+CD25+Foxp3+ regulatory T cells (Tregs). Methods: To generate Aß antigen-specific Tregs (Aß+ Tregs), Aß 1-42 peptide was applied in vivo and subsequent in vitro splenocyte culture. After isolating Tregs by magnetic bead based purification method, Aß+ Tregs were adoptively transferred into 3xTg-AD mice via tail vein injection. Therapeutic efficacy was confirmed with behavior test, Western blot, quantitative real-time PCR (qRT-PCR), enzyme-linked immunosorbent assay (ELISA), and immunohistochemistry staining (IHC). In vitro suppression assay was performed to evaluate the suppressive activity of Aß+ Tregs using flow cytometry. Thy1.1+ Treg trafficking and distribution was analyzed to explore the infused Tregs migration into specific organs in an antigen-driven manner in AD mice. We further assessed cerebral glucose metabolism using 18F-FDG-PET, an imaging approach for AD biological definition. Subsequently, we evaluated the migration of Aß+ Tregs toward Aß activated microglia using live cell imaging, chemotaxis, antibody blocking and migration assay. Results: We showed that Aß-stimulated Tregs inhibited microglial proinflammatory activity and modulated the microglial phenotype via bystander suppression. Single adoptive transfer of Aß+ Tregs was enough to induce amelioration of cognitive impairments, Aß accumulation, hyper-phosphorylation of tau, and neuroinflammation during AD pathology. Moreover, Aß-specific Tregs effectively inhibited inflammation in primary microglia induced by Aß exposure. It may indicate bystander suppression in which Aß-specific Tregs promote immune tolerance by secreting cytokines to modulate immune responses during neurodegeneration. Conclusions: The administration of Aß antigen-specific regulatory T cells may represent a new cellular therapeutic strategy for AD that acts by modulating the inflammatory status in AD.

The Current Status of Clinical Research Involving Microneedles: A Systematic Review.

In recent years, a number of clinical trials have been published on the efficacy and safety of drug delivery using microneedles (MNs). This review aims to systematically summarize and analyze the current evidence including the clinical effect and safety of MNs. Three electronic databases, including PubMed, were used to search the literature for randomized controlled trials (RCTs) and clinical controlled trials (CCTs) that evaluated the therapeutic efficacy of MNs from their inception to 28 June 2018. Data were extracted according to the characteristics of study subjects; disorder, types, and details of the intervention (MNs) and control groups; outcome measurements; effectiveness; and incidence of adverse events (AEs). Overall, 31 RCTs and seven CCTs met the inclusion criteria. Although MNs were commonly used in skin-related studies, evaluating the effects of MNs was difficult because many studies did not provide adequate comparison values between groups. For osteoporosis treatment, vaccine, and insulin delivery studies, MNs were comparable to or more effective than the gold standard. Regarding the safety of MNs, most AEs reported in each study were minor (grade 1 or 2). A well-designed RCT is necessary to clearly evaluate the effectiveness of MNs in the future.

Bee Venom Phospholipase A2 Induces Regulatory T Cell Populations by Suppressing Apoptotic Signaling Pathway.

Bee venom phospholipase A2 is a lipolytic enzyme in bee venom that catalyzes hydrolysis of the sn-2 ester bond of membrane phospholipids to produce free fatty acid and lysophospholipids. Current evidence suggests that bee venom phospholipase A2 (bvPLA2) induces regulatory T cell expansion and attenuates several immune system-related diseases, including Alzheimer's disease. The induction of Treg cells is directly mediated by binding to mannose receptors on dendritic cells. This interaction induces the PGE2-EP2 signaling pathway, which promotes Treg induction in CD4+ T cells. In this study, we investigated the effects of bvPLA2 treatment on the apoptotic signaling pathway in Treg populations. Flow cytometry was performed to identify early apoptotic cells. As a result, early apoptotic cells were dramatically decreased in bvPLA2-treated splenocytes, whereas rapamycin-treated cells showed levels of apoptotic cells similar to those of PBS-treated cells. Furthermore, bvPLA2 treatment increased expression of anti-apoptotic molecules including CTLA-4 and PD-1. The survival rate increased in bvPLA2-treated Tregs. Our findings indicate that bvPLA2-mediated modulation of apoptotic signaling is strongly associated with the Treg induction, which exhibits protective effects against various immune-related diseases. To our knowledge, this study is the first to demonstrate that bvPLA2 is the major bee venom (BV) compound capable of inducing Treg expansion through altering apoptotic signal.

The Methodological Trends of Traditional Herbal Medicine Employing Network Pharmacology.

Natural products, including traditional herbal medicine (THM), are known to exert their therapeutic effects by acting on multiple targets, so researchers have employed network pharmacology methods to decipher the potential mechanisms of THM. To conduct THM-network pharmacology (THM-NP) studies, researchers have employed different tools and databases for constructing and analyzing herb-compound-target networks. In this study, we attempted to capture the methodological trends in THM-NP research. We identified the tools and databases employed to conduct THM-NP studies and visualized their combinatorial patterns. We also constructed co-author and affiliation networks to further understand how the methodologies are employed among researchers. The results showed that the number of THM-NP studies and employed databases/tools have been dramatically increased in the last decade, and there are characteristic patterns in combining methods of each analysis step in THM-NP studies. Overall, the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) was the most frequently employed network pharmacology database in THM-NP studies. Among the processes involved in THM-NP research, the methodology for constructing a compound-target network has shown the greatest change over time. In summary, our analysis describes comprehensive methodological trends and current ideas in research design for network pharmacology researchers.

Sanguiin H-11 from Sanguisorbae radix protects HT22 murine hippocampal cells against glutamate-induced death.

Excessive glutamate level induces neuronal death in acute brain injuries and chronic neurodegenerative diseases. Natural compounds from medicinal and food plants have been attracting interest as a treatment for neurological disorders. Sanguiin H-11 (SH-11), a hydrolysable ellagitannin, inhibits neutrophil movement and nitric oxide -production. However, its neuroprotective effect has not been studied. Therefore, the present study examined the protective effect of SH-11 from Sanguisorbae radix and its mechanism against glutamate-induced death in HT22 cells. Our results showed that SH-11 possessed a strong antioxidant activity and prevented glutamate-induced death in HT22 cells. As a strong antioxidant, SH-11 significantly reduced glutamate-induced increases in intracellular reactive oxygen species accumulation and calcium ion influx. Western blotting analysis showed that glutamate-induced phosphorylation of mitogen-activated protein kinases (MAPKs), including extracellular signal-related kinases 1/2, c-Jun N-terminal kinase, and p38, was significantly decreased by SH-11. Furthermore, SH-11 significantly decreased the number of annexin V-positive HT22 cells, which is indicating apoptotic cell death. In conclusion, our results suggested that SH-11 exerted a potent neuroprotective activity against glutamate-mediated apoptotic cell death by inhibiting oxidative stress-mediated MAPK activation.

Bee venom phospholipase A2 ameliorates Alzheimer's disease pathology in Aß vaccination treatment without inducing neuro-inflammation in a 3xTg-AD mouse model.

Alzheimer's disease (AD) is the most common form of dementia and is characterized by an imbalance between the production and clearance of amyloid-beta (Aß) and tau proteins. Although vaccination against Aß peptide results in a dramatic reduction in Aß pathology in experimental mouse models, the initial clinical trial for an active Aß vaccine was halted early due to the development of acute meningoencephalitis in 6% of the immunized patients, which likely involved a T-cell mediated pro-inflammatory response. In this study, we aimed to determine whether bee venom phospholipase A2 (bvPLA2) treatment would induce Tregs and ameliorate AD pathology without unwanted T cell-mediated inflammation. First, we investigated the effects of bvPLA2 on the inflammatory infiltration caused by Aß vaccination. Inflammatory aggregates of CD3+ T lymphocytes and macrophages were found in the brains and spinal cords of mice treated with Aß. However, administration of bvPLA2 dramatically eliminated central nervous system inflammation following Aß immunization. In AD model mice (3xTg-AD mice), bvPLA2 administration significantly ameliorated cognitive deficits and reduced Aß burdens in the brains of Aß-vaccinated 3xTg-AD mice. Additionally, we examined brain glucose metabolism using positron emission tomography with 18F-2 fluoro-2-deoxy-D-glucose. Cerebral glucose uptake was considerably higher in the brains of Aß-vaccinated 3xTg-AD mice that received bvPLA2 than those that did not. The present study suggests that the modulation of Treg populations via bvPLA2 treatment may be a new therapeutic approach to attenuate the progression of AD in conjunction with Aß vaccination therapy without an adverse inflammatory response.

Bee venom phospholipase A2 ameliorates motor dysfunction and modulates microglia activation in Parkinson's disease alpha-synuclein transgenic mice.

α-Synuclein (α-Syn) has a critical role in microglia-mediated neuroinflammation, which leads to the development of Parkinson's disease (PD). Recent studies have shown that bee venom (BV) has beneficial effects on PD symptoms in human patients or 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) toxin-induced PD mice. This study investigated whether treatment with BV-derived phospholipase A2 (bvPLA2) would improve the motor dysfunction and pathological features of PD in human A53T α-Syn mutant transgenic (A53T Tg) mice. The motor dysfunction of A53T Tg mice was assessed using the pole test. The levels of α-Syn, microglia and the M1/M2 phenotype in the spinal cord were evaluated by immunofluorescence. bvPLA2 treatment significantly ameliorated motor dysfunction in A53T Tg mice. In addition, bvPLA2 significantly reduced the expression of α-Syn, the activation and numbers of microglia, and the ratio of M1/M2 in A53T Tg mice. These results suggest that bvPLA2 could be a promising treatment option for PD.

Protective Effect of Coriolus versicolor Cultivated in Citrus Extract Against Nitric Oxide-Induced Apoptosis in Human Neuroblastoma SK-N-MC Cells.

Nitric oxide (NO) is a reactive free radical and a messenger molecule in many physiological functions. However, excessive NO is believed to be a mediator of neurotoxicity. The medicinal plant Coriolus versicolor is known to possess anti-tumor and immune-potentiating activities. In this study, we investigated whether Coriolus versicolor possesses a protective effect against NO donor sodium nitroprusside (SNP)-induced apoptosis in the human neuroblastoma cell line SK-N-MC. We utilized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, flow cytometry, 4,6-diamidino-2-phenylindole (DAPI) staining, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) assay, DNA fragmentation assay, reverse transcription-polymerase chain reaction (RT-PCR), Western blot analysis, and caspase-3 enzyme activity assay in SK-N-MC cells. MTT assay showed that SNP treatment significantly reduces the viability of cells, and the viabilities of cells pre-treated with the aqueous extract of Coriolus versicolor cultivated in citrus extract (CVE(citrus)) was increased. However, aqueous extract of Coriolus versicolor cultivated in synthetic medium (CVE(synthetic)) showed no protective effect and aqueous citrus extract (CE) had a little protective effect. The cell treated with SNP exhibited several apoptotic features, while those pre-treated for 1 h with CVE(citrus) prior to SNP expose showed reduced apoptotic features. The cells pre-treated for 1 h with CVE(citrus) prior to SNP expose inhibited p53 and Bax expressions and caspase-3 enzyme activity up-regulated by SNP. We showed that CVE(citrus) exerts a protective effect against SNP-induced apoptosis in SK-N-MC cells. Our study suggests that CVE(citrus) has therapeutic value in the treatment of a variety of NO-induced brain diseases.

Anti-diabetic effect of standardized herbal formula PM021 consisting of Mori Folium and Aurantii Fructus on type II diabetic Otsuka Long-Evans Tokushima Fatty (OLETF) rats.

PM021, which consists of two herbal components, Mori Folium and Aurantii Fructus, is routinely used to treat diabetes in Korea. In this study, the anti-diabetic effect of PM021 on an animal model of developing type 2 diabetes of Otsuka Long-Evans Tokushima Fatty (OLETF) rats was investigated. Eight weeks of age male OLETF rats were treated daily with PM021 or vehicle for five months. Specifically, changes in body weight, blood glucose, urine volume, food intake and oral glucose tolerance were measured in rats for five months. The rats in this study were divided into four groups: a Long-Evans Tokushima Otsuka (LETO) rat group, which is a genetic control group for OLETF, that received no treatment; a PM021 treatment group of LETO rats; OLETF rats that received no treatment; and OLETF rats that received PM021 treatment. The results showed that PM021 significantly prevented increases in body weight, blood glucose, and urine and food intake that resulted from the induction of obesity and diabetes. PM021 also improved glucose tolerance in OLETO rats. However, PM021 had no effect on LETO rats, a control group of OLETF rats. Taken together, these findings indicate that PM021 has distinct anti-diabetic effects without any adverse effects or toxicities.

Effects of acupuncture on abdominal leak point pressure and c-Fos expression in the brain of rats with stress urinary incontinence.

Stress urinary incontinence leads to the involuntary loss of urine during abdominal strain caused by sneezing, laughing, and coughing. Acupuncture has been widely used for the treatment and prevention of a variety of diseases in traditional medicine. Acupuncture has also been used to relieve the symptoms of functional disorders of the lower urinary tract. In the present study, we investigated the effect of acupuncture at the Sanyinjiao (SP6) acupoint on stress urinary incontinence in rats. The present results showed that abdominal leak point pressure was decreased in rats with stress urinary incontinence, while acupuncture at the SP6 acupoint significantly enhanced the abdominal leak point pressure. The expression of c-Fos in the pontine micturition center (PMC), ventrolateral periaqueductal gray (vlPAG), and medial preoptic nucleus (MPA) regions was increased by the induction of stress urinary incontinence, and acupuncture at the SP6 acupoint significantly decreased c-Fos expression in these areas. In the present study, we showed that acupuncture has therapeutic effect on the symptoms of stress urinary incontinence, and this effect of acupuncture is associated with modulation of c-Fos expression in the brain.

Aqueous extract of Anemarrhena rhizome increases cell proliferation and neuropeptide Y expression in the hippocampal dentate gyrus on streptozotocin-induced diabetic rats.

In this study, the effects of the aqueous extract of Anemarrhena rhizome on cell proliferation and neuropeptide Y expression in the dentate gyrus of streptozotocin-induced diabetic rats were investigated via immunohistochemistry for 5-bromo-2'-deoxyuridine (BrdU) and neuropeptide Y. The results showed that the treatment with 50 to 200 microg/kg/day for 7 days of the aqueous extract of Anemarrhena rhizome increased new cell formation and neuropeptide Y expression in the dentate gyrus of diabetic rats reduced by the treatment with streptozotocin in rat.

Effects of acupuncture on the intrastriatal hemorrhage-induced caspase3 expression and newly cell birth in rats.

BACKGROUND: Intracerebral hemorrhage is one of the most devastating types of stroke. Caspases are essential players in apoptotic cell death both as initiators and executioners. The v-Fos FBJ murine osteosarcoma viral oncogene homolog (Fos, c-Fos) is an immediate early gene, and Fos expression is sometimes used as a marker for stimuli-induced changes in the metabolic activity of neurons. The expressions of caspase3 and Fos are enhanced with neuroregeneration and with neuronal cell death, respectively. Cells proliferation the dentate gyrus of adult rodents is enhanced by certain pathologic events as seizures and ischemic insult, and such up-regulation of cell proliferation occurring during pathologic situations is thought to be a compensatory response to lesion-induced cell death in the brain. In the present study, we investigated the effects of acupuncture on the intrastriatal hemorrhage-induced caspase3 expression in the striatum and on the Fos expression and cell proliferation in the dentate gyrus of rats. METHODS: For this study, immunohistochemistry for caspase3, Fos and 5-bromo-2'-deoxyuridine (BrdU) was performed. RESULTS: Caspase3 expression in the striatum was increased by intrastriatal hemorrhage. Fos expression and cell proliferation in the dentate gyrus of rats with intracerebral hemorrhage were also increased. Acupunctural treatment, especially at the ST36 acupoint, suppressed the intracerebral hemorrhage-induced caspase3 expression in the stratum, and it also inhibited expression of Fos and cell proliferation in the dentate gyrus. CONCLUSION: In the present study, we have shown that acupuncture treatment has a neuroprotective effect on intrastrstriatal hemorrhage-induced neuronal cell death, and this suggests that acupuncture can aid in the recovery of the central nervous system following stroke.