Naodesheng (NDS) formula, which consists of Rhizoma Chuanxiong, Lobed Kudzuvine, Carthamus tinctorius, Radix Notoginseng, and Crataegus pinnatifida, is widely applied for the treatment of cardio/cerebrovascular ischemic diseases, ischemic stroke, and sequelae of cerebral hemorrhage, etc. At present, the studies on NDS formula for Alzheimer's disease (AD) only focus on single component of this prescription, and there is no report about the synergistic mechanism of the constituents in NDS formula for the potential treatment of dementia. Therefore, the present study aimed to predict the potential targets and uncover the mechanisms of NDS formula for the treatment of AD. Firstly, we collected the constituents in NDS formula and key targets toward AD. Then, drug-likeness, oral bioavailability, and blood-brain barrier permeability were evaluated to find drug-like and lead-like constituents for treatment of central nervous system diseases. By combining the advantages of machine learning, molecular docking, and pharmacophore mapping, we attempted to predict the targets of constituents and find potential multi-target compounds from NDS formula. Finally, we built constituent-target network, constituent-target-target network and target-biological pathway network to study the network pharmacology of the constituents in NDS formula. To the best of our knowledge, this represented the first to study the mechanism of NDS formula for potential efficacy for AD treatment by means of the virtual screening and network pharmacology methods.
Alzheimer Disease ; drug therapy ; pathology ; physiopathology ; Autoanalysis ; Biological Availability ; Biomarkers ; Biomarkers, Pharmacological ; Databases, Chemical ; Drug Combinations ; Drug Discovery ; methods ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; therapeutic use ; Humans ; Machine Learning ; Molecular Docking Simulation ; Neural Networks, Computer ; Peptide Fragments ; chemistry ; Permeability

Alzheimer's disease (AD) is one of the major neurodegenerative disorders of the elderly, which is characterized by the accumulation and deposition of amyloid-beta (Aβ) peptide in human brains. Oxidative stress and neuroinflammation induced by Aβ in brain are increasingly considered to be responsible for the pathogenesis of AD. The present study aimed to determine the protective effects of walnut peptides against the neurotoxicity induced by Aβ25-35 in vivo. Briefly, the AD model was induced by injecting Aβ25-35 into bilateral hippocampi of mice. The animals were treated with distilled water or walnut peptides (200, 400 and 800 mg/kg, p.o.) for five consecutive weeks. Spatial learning and memory abilities of mice were investigated by Morris water maze test and step-down avoidance test. To further explore the underlying mechanisms of the neuroprotectivity of walnut peptides, the activities of superoxide dismutase (SOD), glutathione (GSH), acetylcholine esterase (AChE), and the content of malondialdehyde (MDA) as well as the level of nitric oxide (NO) in the hippocampus of mice were measured by spectrophotometric method. In addition, the levels of 8-hydroxy-2'-deoxyguanosine (8-OHdG), tumor necrosis factor-α (TNF-α), interleukin 1β (IL-1β) and IL-6 in the samples were determined using ELISA. The hippocampal expressions of inducible nitric oxide synthase (iNOS) and nuclear factor κB (NF-κB) were evaluated by Western blot analysis. The results showed that walnut peptides supplementation effectively ameliorated the cognitive deficits and memory impairment of mice. Meanwhile, our study also revealed effective restoration of levels of antioxidant enzymes as well as inflammatory mediators with supplementation of walnut peptides (400 or 800 mg/kg). All the above findings suggested that walnut peptides may have a protective effect on AD by reducing inflammatory responses and modulating antioxidant system.
Acetylcholinesterase ; metabolism ; Alzheimer Disease ; drug therapy ; etiology ; Amyloid beta-Peptides ; toxicity ; Animals ; Female ; Glutathione ; metabolism ; Hippocampus ; drug effects ; metabolism ; Interleukins ; metabolism ; Juglans ; chemistry ; Male ; Malondialdehyde ; metabolism ; Maze Learning ; Memory Disorders ; drug therapy ; etiology ; Mice ; NF-kappa B ; metabolism ; Neuroprotective Agents ; pharmacology ; therapeutic use ; Nitric Oxide ; metabolism ; Peptide Fragments ; toxicity ; Peptides ; pharmacology ; therapeutic use ; Plant Extracts ; pharmacology ; therapeutic use ; Superoxide Dismutase ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo investigate the effects of exogenous hydrogen sulfide (H2S) on β-site APP cleaving enzyme 1 (BACE-1) and β-amyloid peptide (Aβ) metabolism in primary culture of neurons under high-glucose condition.

METHODSThe cortical neurons in primary culture under normal and high glucose (60 mmol/L) conditions for 24 h were exposed to 25, 50 and 100 µmol/L NaHS. Aβ1-42 concentration in the cell culture was measured by ELISA, and BACE-1 mRNA and protein levels were detected by fluorescent quantitative real-time PCR and Western blotting, respectively.

RESULTSCompared with the neurons cultured in normal glucose, the neurons exposed to high glucose showed significantly increased Aβ1-42 concentration and BACE-1 mRNA and protein expressions (P<0.05). Exposure to 25, 50 and 100 µmol/L NaHS significantly decreased Aβ1-42 concentration and BACE-1 mRNA and protein expressions in the high-glucose cell culture (P<0.05).

CONCLUSIONNeurons exposed to high glucose exhibit increased Aβ1-42 levels and BACE-1 mRNA and protein expressions, which can be concentration-dependently decreased by NaHS.

Amyloid Precursor Protein Secretases ; metabolism ; Amyloid beta-Peptides ; metabolism ; Animals ; Aspartic Acid Endopeptidases ; metabolism ; Cells, Cultured ; Culture Media ; chemistry ; Glucose ; chemistry ; Hydrogen Sulfide ; pharmacology ; Neurons ; drug effects ; metabolism ; Peptide Fragments ; metabolism ; Rats ; Rats, Sprague-Dawley

A major barrier to the use of antimicrobial peptides as antibiotics is the toxicity or ability to lyse eukaryotic cells. In this study, a 26-residue amphipathic α-helical antimicrobial peptide A12L/A20L (Ac-KWKSFLKTFKSLKKTVLHTLLKAISS-amide) was used as the framework to design a series of D- and L-diastereomeric peptides and study the relationships of helicity and biological activities of α-helical antimicrobial peptides. Peptide helicity was measured by circular dichroism spectroscopy and demonstrated to correlate with the hydrophobicity of peptides and the numbers of D-amino acid substitutions. Therapeutic index was used to evaluate the selectivity of peptides against prokaryotic cells. By introducing D-amino acids to replace the original L-amino acids on the non-polar face or the polar face of the helix, the hemolytic activity of peptide analogs have been significantly reduced. Compared to the parent peptide, the therapeutic indices were improved of 44-fold and 22-fold against Gram-negative and Gram-positive bacteria, respectively. In addition, D- and L-diastereomeric peptides exhibited lower interaction with zwitterionic eukaryotic membrane and showed the significant membrane damaging effect to bacterial cells. Helicity was proved to play a crucial role on peptide specificity and biological activities. By simply replacing the hydrophobic or the hydrophilic amino acid residues on the non-polar or the polar face of these amphipathic derivatives of the parent peptide with D-amino acids, we demonstrated that this method could have excellent potential for the rational design of antimicrobial peptides with enhanced specificity.
Anti-Infective Agents ; chemistry ; pharmacology ; Circular Dichroism ; Drug Design ; Erythrocytes ; drug effects ; Gram-Negative Bacteria ; drug effects ; Gram-Positive Bacteria ; drug effects ; Hemolysis ; drug effects ; Humans ; Peptide Fragments ; chemistry ; pharmacology ; Protein Structure, Secondary ; Structure-Activity Relationship ; Substrate Specificity

OBJECTIVE: To prepare Arg-Gly-Asp (RGD) and cell penetrating peptide TAT co-modified paclitaxel loaded liposome (RGD/TAT-LP-PTX) for MCF-7 cell inhibition. METHODS: The co-modified liposome was prepared by film-ultrasonic method. The appearance, particle size and zeta potential were evaluated. The cellular uptake by MCF-7 cells in vitro was used to evaluate the targeting efficiency. The anti-proliferation efficiency of RGD/TAT-LP-PTX was evaluated by MTT assay. Tumor spheroids were used to evaluate anti-tumor ability of RGD/TAT-LP-PTX in vitro. RESULTS: The particle diameter of the co-modified liposome was (138.8 ± 12.4) nm with the Zeta potential of (25.85 ± 2.75) mV. The entrapment efficiency of PTX was 88.3%. The RGD/TAT-LP uptaken by MCF-7 cells at 4 h was 1.79 times higher than that at 2 h. The co-modified liposome uptaken by MCF-7 cells was 2.25 and 2.72 times higher than that of TAT-LP and RGD-LP, respectively. The anti-proliferation rate of RGD/TAT-LP-PTX increased with time. The inhibition rate of RGD/TAT-LP-PTX for MCF-7 cells at 48 h was 1.78 times higher than that at 24 h. The MTT assay demonstrated the cell viability of RGD/TAT-LP-PTX was 1.65, 1.82 and 2.55 times higher than that of TAT-LP-PTX, RGD-LP-PTX and LP-PTX, respectively. CONCLUSION Co-modified liposome may serve as a promising breast cancer delivery system for antitumor drugs.
Antineoplastic Agents ; pharmacology ; Breast Neoplasms ; Cell Survival ; Humans ; Liposomes ; MCF-7 Cells ; drug effects ; Oligopeptides ; chemistry ; Paclitaxel ; pharmacology ; Particle Size ; Peptide Fragments ; chemistry ; Spheroids, Cellular ; drug effects

Calreticulin (CRT) is a multifunctional molecule in both intracellular and extracellular environment. We have previously found that a recombinant CRT fragment (rCRT/39-272) could modulate T cell-mediated immunity in mice via activation and expansion of CD1d(hi)CD5⁺ B cells as well as induction of CRT-specific regulatory antibodies. Antibody secreting cells (ASCs) are terminally differentiated B cells responsible for producing antibodies to participate in positive immune response as well as immune regulation. In this study, we demonstrate that rCRT/39-272 differentiates murine CD1d(hi)CD5⁺ B cells into ASCs marked by increased expression of plasma cell-associated transcription factors and production of polyreactive antibodies against DNA and CRT in vitro. Intraperitoneal administration of rCRT/39-272 augmented differentiation of CD1d(hi)CD5⁺ B cells into ASCs in naïve mice or mice with experimental autoimmune encephalomyelitis. Thus, we propose that ASC differentiation and subsequent antibody production of CD1d(hi)CD5⁺ B cells are key steps in CRT-mediated immunoregulation on inflammatory T cell responses.
Animals ; Antigens, CD1d ; metabolism ; Autoantibodies ; biosynthesis ; B-Lymphocytes ; cytology ; drug effects ; immunology ; metabolism ; CD5 Antigens ; metabolism ; Calreticulin ; chemistry ; Cell Differentiation ; drug effects ; Encephalomyelitis, Autoimmune, Experimental ; immunology ; Humans ; Mice ; Peptide Fragments ; chemistry ; pharmacology ; Solubility

Tyrosinase is a key enzyme related to skin pigmentation disorders of elderly people, while self-aggregation of the amyloid-beta peptide, Abeta42, has been considered as a key event in the pathogenesis of Alzheimer's disease (AD). The present study was undertaken to investigate the inhibitory effects of 20 samples from Rhodiola species on tyrosinase and Abeta42 aggregation, and to isolate their corresponding bioactive components. The results demonstrated that the oligomeric proanthocyanidins (OPCs) commonly found in Rhodiola species were the major bioactive components corresponding to their anti-tyrosinase and anti-Abeta42 aggregation bioactivities. Salidroside, a representative compound of Rhodiola plants, proved not to be active in the present studies.
Amyloid beta-Peptides ; antagonists & inhibitors ; metabolism ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Glucosides ; isolation & purification ; pharmacology ; Monophenol Monooxygenase ; antagonists & inhibitors ; metabolism ; Peptide Fragments ; antagonists & inhibitors ; metabolism ; Phenols ; isolation & purification ; pharmacology ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Proanthocyanidins ; isolation & purification ; pharmacology ; Rhodiola ; chemistry

OBJECTIVETo observe the injury in rat primary cultured neurons induced by Abeta(1-40) and the protective effects of combination of ginseng and ginko extracts.

METHODPrimary neurons were induced by Abeta(1-40) to establish the cell model of toxic injury. Using flow cytometry with Annexin V-FITC/PI double staining, MTP assay, transmission electron microscopy and Western blot, the appropriate concentration and duration of AP for cell model establishment were determined. The effects of extracts of ginseng and ginko (EGGB)on cellular proliferative activity, apoptotic rate, ultrastructure and caspase-3 expression were detected.

RESULTThe apoptotic rate was increased significantly after neurons were induced by 1 micromol x L(-1) Abeta(-40) for 24 h (P < 0.01). EGGB (5, 50 mg L(-1)) significantly enhanced the proliferative activity (P < 0.05). Meanwhile, EGGB (50 mg L(-1)) inhibited neuronal apoptosis and caspase-3 overexpression and improved cellular ultrastructure remarkably (P < 0.05, P < 0.01).

CONCLUSIONAbeta(1-40) could significantly induce primary cultured neurons to apoptosis in vitro. EGGB showed beneficial neuroprotective effects against neuronal apoptosis, which might be due to improving the structures of neuron and its subcellular organelles, enhancing cellular proliferative activity and inhibiting caspase-3 overexpression in neurons.

Alzheimer Disease ; drug therapy ; Amyloid beta-Peptides ; pharmacology ; Animals ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Proliferation ; drug effects ; Cells, Cultured ; Drug Interactions ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Gene Expression Regulation, Enzymologic ; drug effects ; Ginkgo biloba ; chemistry ; Male ; Neurons ; cytology ; drug effects ; metabolism ; ultrastructure ; Panax ; chemistry ; Peptide Fragments ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Time Factors

OBJECTIVETo observe the effects of hippocampal Abeta42 deposition on the expression of inflammatory cytokines and phosphorylated MAPK signal molecules as well as the intervention of AD by total glucosides of paeony (TGP).

METHOD12 week-old female SD rats were stereotactic injected one-time with a fibrillar Abeta42 positioning hippocampus to replicate AD pathology model and interfered with TGP. The expression of inflammatory cytokines and phosphorylated MAPK pathway signaling molecules were observed by immunohistochemistry (SABC), and SABC images were analyzed by image analysis software.

RESULTCompared with the control group, the IL-1beta, IL-6 and p-p38, p-JNK, p-MEK3/6 positive stained areas of AD pathology model group increased and their staining intensity decreased (the protein expression quantity inversely proportional to the staining intensity), while the IL-1beta, IL-6 and p-p38, p-JNK, p-MEK3/6 positive stained areas of the treatment groups decreased and their staining intensity increased compared with AD pathology model group.

CONCLUSIONAbeta42 deposition in hippocampus can induce the brain inflammation and the over-expression of IL-1beta, IL-6 and p-p38, p-JNK, p-MEK3/6. Inhibiting the over-expression of inflammatory cytokines and phosphorylated MAPK signaling molecules may be a major antagonistic mechanism of TGP against AD.

Alzheimer Disease ; drug therapy ; Amyloid beta-Peptides ; metabolism ; toxicity ; Animals ; Cytokines ; analysis ; Female ; Glucosides ; pharmacology ; therapeutic use ; Hippocampus ; metabolism ; JNK Mitogen-Activated Protein Kinases ; metabolism ; MAP Kinase Signaling System ; drug effects ; Mitogen-Activated Protein Kinases ; metabolism ; Paeonia ; chemistry ; Peptide Fragments ; metabolism ; toxicity ; Phosphorylation ; Rats ; Rats, Sprague-Dawley ; p38 Mitogen-Activated Protein Kinases ; metabolism

To investigate the angiotensin I-converting enzyme (ACE) inhibitory activity of beta-chain hemoglobin fragments, 17 fragments were synthesized by microwave-assisted solid-phase synthesis method. Wang resin or Trt(2-Cl) resin, Fmoc and HBTU-HOBt were used as solid carrier, N-terminal amino acid protecting groups and coupling reagents, respectively. The ACE inhibitory, alpha-glucosidase inhibitory, antibacterial and antitumor activities of the synthesized fragments were assayed. In vitro, Val-Val-Tyr-Pro-Trp-Thr showed high ACE inhibitory activity (IC50 = 7.42 micromol x L(-1)). The results indicate that there are two active sites in Val-Val-Tyr-Pro-Trp-Thr-Gln-Arg-Phe, one consists of Val-Val-, and the other -Gln-Arg-Phe. Peptides showed high ACE inhibitory activity when the N-terminal was hydrophobic amino acid such as Val and C-terminal tripeptide contained Phe, Trp or Arg. Some of the fragments showed low a-glucosidase inhibitory activity. No antibacterial activity or antitumor activity was detected in vitro. The results indicate that these peptides have a potential antihypertensive effect and possible application in the treatment of hypertension.
Amino Acid Sequence ; Angiotensin-Converting Enzyme Inhibitors ; pharmacology ; Anti-Bacterial Agents ; pharmacology ; Antihypertensive Agents ; pharmacology ; Antineoplastic Agents ; pharmacology ; Cell Line, Tumor ; Glycoside Hydrolase Inhibitors ; Humans ; Peptide Fragments ; chemical synthesis ; chemistry ; pharmacology ; Peptidyl-Dipeptidase A ; drug effects ; Solid-Phase Synthesis Techniques ; methods ; alpha-Glucosidases ; drug effects ; beta-Globins ; chemical synthesis ; chemistry ; pharmacology