The study reports the detection of neuroprotective effect of 10 kinds of coumarin derivatives and explores their possible mechanism. MTT method was used to screen the neuroprotective effect of 10 coumarin derivatives on neurotoxic agents (Aβ25-35 and rotenone) or OGD (oxygen-glucose deprivation). A compound with better protective effect was obtained. Then the effect of this compound on neurotoxic agents on PC12 was detected by the morphological observation. Furthermore, the effect of compound 3 on microglia with lipopolysaccharide (LPS) induced inflammation was detected. And the inflammatory factor was tested. Finally, direct free radical scavenging ability was detected. Compound 3 was found to be the best compound through three neurons toxic models. Not only compound 3 ameliorated cell viability reduced by three neurons toxic models, but also significantly inhibited the production of inflammatory factor (TNF-α and IL-1β). And its free radical scavenging ability is very good, especially the effect on superoxide anion, which is comparable with vitamin C. The significant scavenging effect of compound 3 on superoxide anion might be the mechanism of the neuroprotection. Compound 3 as a potential neural cell protective agent merits further investigation.
Animals ; Coumarins ; chemistry ; Free Radical Scavengers ; chemistry ; Inflammation ; Microglia ; drug effects ; Neurons ; drug effects ; Neuroprotective Agents ; chemistry ; PC12 Cells ; Rats

The amyloid β-protein (Aβ)-induced disturbance of intracellular calcium homeostasis has been regarded as the final route whereby Aβ insults neurons. However, the mechanism of Aβ-induced Ca(2+) overloading is still unclear so far. Especially, it remains to be clarified whether nicotinic acetylcholine receptors (nAChRs) are involved in the Aβ-induced elevation of intracellular calcium concentration ([Ca(2+)](i)). In the present study, we observed the effects of Aβ fragments 25-35 (Aβ(25-35)) and 31-35 (Aβ(31-35)) on [Ca(2+)](i) in primary cultured rat cortical neurons using laser-scanning confocal calcium imaging technique, and investigated its probable cholinergic mechanism. The results showed that: (1) Both Aβ(25-35) and Aβ(31-35) induced similar and significant [Ca(2+)](i) elevation in a concentration-dependent manner, and no statistical difference was found between the effects of both peptides; (2) The reverse peptide of Aβ(31-35), i.e. Aβ(35-31), had no effect on [Ca(2+)](i) elevation; (3) Mecamylamine (MCA), a non-specific nAChRs antagonist, significantly and dose-dependently blocked the [Ca(2+)](i) elevation induced by Aβ(25-35) or Aβ(31-35) (4) Dihydro-β-erythroidine (D-β-E), a specific α4β2 subtype nAChRs antagonist, also significantly inhibited the [Ca(2+)](i) elevation induced by Aβ(25-35) and Aβ(31-35), but the effect was weaker than the effect of MCA at the same concentration. These results indicate that Aβ(31-35) may be a shorter active sequence in full length of Aβ molecule, and the overactivation of nAChRs, including α4β2 subtype, may be, at least partly, responsible for the Aβ-induced elevation of [Ca(2+)](i) in cultured rat cortical neurons. Thus, the present study suggests a new potential target of Aβ in the brain, and provides a new insight into the mechanisms by which Aβ impairs the cognitive function in Alzheimer's disease.
Amyloid beta-Peptides ; chemistry ; Animals ; Calcium ; metabolism ; Cells, Cultured ; Neurons ; metabolism ; Peptide Fragments ; chemistry ; Rats ; Receptors, Nicotinic ; metabolism

The olfactory perception is the process that the olfactory receptor is activated by odorous molecules, which induce the transduction of signal in the cell and the chemical information is transduced into electrical impulses. After the changed signal is transmitted to the brain, the whole perception process completes. OR gene belongs to the multigene family. The coded olfactory receptor proteins belong to the G-protein-coupled receptor (GPCR) superfamily and therefore are invariably seven-transmembrane domain(7TM) protein. Olfactory receptor protein plays an important role in olfactory perception and signal transduction process.
Animals ; Humans ; Olfactory Receptor Neurons ; metabolism ; physiology ; Receptors, Odorant ; chemistry ; genetics ; physiology ; Signal Transduction

OBJECTIVETo study the relationship between substance P (SP) and/or calcitonin gene-related peptide (CGRP) immunoreactive neurons in dorsal root ganglia (DRG) and the transmission of nociception in the penile frenulum of rats.

METHODSThe fluoro-gold (FG) retrograde tracing method was used to trace the origin of nerve terminals in the penile frenulum of rats. And SP and/or CGRP immunofluorescence labeling was employed to detect the distribution of SP and/or CGRP immunoreactive neurons in DRG.

RESULTSFG retrograde tracing showed that the FG retrolabeled neurons were localized in L6-DRG and S1-DRG. SP and/or CGRP immunofluorescence labeling indicated that a large number of DRG neurons were SP- and CGRP-immunoreactive, different in size, bright red and bright green respectively in color, and arranged in rows or spots among nerve bundles. All the FG/SP and FG/CGRP double-labeled neurons were medium or small-sized. One third of the FG-labeled neurons were SP-immunoreactive, and a half of them CGRP-immunoreactive in L6-DRG and S1-DRG respectively. The FG/SP/CGRP-labeled neurons accounted for one fifth of the FG retro labeled neurons.

CONCLUSIONSP- and CGRP-immunoreactive neurons in L6-DRG and SI-DRG of rats may be involved in the transmission of nociception in rat penile frenulum.

Animals ; Calcitonin Gene-Related Peptide ; analysis ; Ganglia, Spinal ; chemistry ; cytology ; Male ; Microscopy, Fluorescence ; Neurons ; chemistry ; physiology ; Neurons, Afferent ; chemistry ; physiology ; Penis ; innervation ; Rats ; Rats, Sprague-Dawley ; Substance P ; analysis

OBJECTIVETo study the level of brain-derived neurotrophic factor (BDNF) in the microenvironment of the neuron-astrocyte co-culture system by Mg2+-free-induced seizure-like discharge and analyze the source of BDNF.

METHODSHippocampal neurons (N) of fetal rats and astrocytes (AST) of neonatal rats were purified and divided into four groups, included control N (Con N) group, Mg2+-free treated N (Mg2+-free N) group, control N+AST co-culture (Con N+AST) group and Mg2+-free treated N+AST co-culture (Mg2+-free N+AST) group. The Mg2+-free treated groups were exposed to Mg2+-free media for 3 hrs to induce a repeated spontaneous seizure-like discharge. The level of BDNF in each group at different time points was measured using ELISA.

RESULTSThe cellular morphous of AST changed in the Mg2+-free N+AST group at 48 hrs. Neuronal epileptiform activity was observed in the Mg2+-free media at 3 hrs, and continued to exist until the microenvironment returned to normal for 72 hrs. The BDNF level increased at 24 hrs and 48 hrs in the Con N+AST group compared with the control N group (P<0.05). Compared with Con N+AST group, BDNF level increased at 12, 24 and 48 hrs in the Mg2+-free N+AST group, especially at 12 and 24 hrs (P<0.01). There were no significant differences in the level of BDNF between the Con and Mg2+-free N groups. Compared with Mg2+-free N group, BDNF level increased at 24 hrs in the Mg2+-free N+AST group (P<0.05).

CONCLUSIONSThe results of the experiment suggest that BDNF in the Con N+AST group might be excreted from both N and AST, but chiefly from N. Activated AST may be the main source for increasing BDNF in the Mg2+-free N+AST group.

Animals ; Astrocytes ; chemistry ; Brain-Derived Neurotrophic Factor ; analysis ; Cellular Microenvironment ; Coculture Techniques ; Female ; Hippocampus ; chemistry ; Magnesium Deficiency ; metabolism ; Male ; Neurons ; chemistry ; Rats ; Rats, Wistar ; Seizures ; metabolism

OBJECTIVETo study the relationship between the protection of Ginsenoside(GS) for spinal cells and nitric oxide (NO).

METHODSpinal cells were cultured in vitro, the model of peripheral nerve was established by scarifying the cells, and NO was measured by Griess method.

RESULTNO in injury group was high than that in noninjury group and NO in group cultured by GS was less than that in group cultured by common medium.

CONCLUSIONNO increases when peripheral nerve is injuried, and the protective effect of GS on spinal cells may be through inhibiting NO release.

Animals ; Cells, Cultured ; Fetus ; Ginsenosides ; isolation & purification ; pharmacology ; Motor Neurons ; cytology ; drug effects ; metabolism ; Neurons, Afferent ; cytology ; drug effects ; metabolism ; Neuroprotective Agents ; pharmacology ; Nitric Oxide ; metabolism ; Panax ; chemistry ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley ; Spinal Cord ; cytology ; metabolism

PURPOSE: Apoptosis is active cell death which plays an important role in developing normal tissues. Various conditions such as genetic defects, drugs, ischemia or infections are known to induce apoptosis. We studied the effect of maternal infection on fetal brain development during pregnancy. METHODS: We treated 46 C3H pregnant mice with lipopolysaccharide (LPS) or phosphat-buffered saline and observed the changes in apoptosis and expression of bcl-2, bcl-xS, bax. The fetal brain tissues were removed 1-48 hours after LPS treatment. The number of apoptosis per 100 neurons and glial cells was counted in H&E stained tissue and was analyzed statistically. Immunohistochemical staining with primary antibodies of bcl-2, bcl-xS, bax was done and their expression was classified by the degree of staining. RESULTS: The number of apoptosis was increased significantly in both neurons and glial cells of LPS-treated group and its degree of staining was more remarkable in glial cells. Immunohisto chemistry for bcl-2, bcl-xS, bax oncoprotein revealed mildly decreased expression of bcl-2 and markedly increased expression of bax in both neurons and glial cells, but it was more remarkable in glial cells. Immunochemistry for bcl-xS revealed no expression in neurons and minimal expression of bcl-xS in glial cells in both study groups. CONCLUSOIN: We observed an increase in the number of apoptosis, mildly decreased expression of bcl-2 and markedly increased expression of bax in both neurons and glial cells of fetal brain after treating pregnant mice with LPS. Maternal infection during pregnancy may have profound effects on developing fetal brain.
Animals ; Antibodies ; Apoptosis* ; bcl-2-Associated X Protein* ; Brain* ; Cell Death ; Chemistry ; Immunochemistry ; Ischemia ; Mice* ; Neuroglia ; Neurons ; Pregnancy

The present study was aimed to investigate the role of cerebrospinal fluid-contacting nucleus (CSF-CN) neurons in modulation of inflammatory pain and underlying mechanism. The inflammatory pain model was made by subcutaneous injection of the complete Freund's adjuvant (CFA) into the left hind paw of rats. The phosphorylation level of PKC (p-PKC) was examined by Western blot. Thermal withdrawal latency (TWL) of the rats was measured to assess inflammatory pain. The results showed that, compared with the sham controls, the inflammatory pain model rats showed shortened TWL on day 1, 3, and 7 after CFA injection, as well as increased level of p-PKC in CSF-CN neurons at 24 h after CFA injection. The administration of GF109203X, a PKC inhibitor, into lateral ventricle decreased the level of p-PKC protein expression and increased TWL in the model rats. These results suggest that blocking the PKC pathway in CSF-CN neurons may be an effective way to reduce or eliminate the inflammatory pain.
Animals ; Freund's Adjuvant ; Inflammation ; enzymology ; Neurons ; enzymology ; Pain ; enzymology ; Phosphorylation ; Protein Kinase C ; cerebrospinal fluid ; chemistry ; Rats ; Rats, Sprague-Dawley

Voltage-gated sodium channels (VGSCs), which are widely distributed in the excitable cells, are the primary mediators of electrical signal amplification and propagation. They play important roles in the excitative conduction of the neurons and cardiac muscle cells. The abnormalities of the structures and functions of VGSCs can change the excitability of the cells, resulting in a variety of diseases such as neuropathic pain, epilepsy and arrhythmia. At present, some voltage-gated sodium channel blockers are used for treating those diseases. In the recent years, several neurotoxins have been purified from the venom of the animals, which could inhibit the current of the voltage-gated sodium channels. Usually, these neurotoxins are compounds or small peptides that have been further designed and modified for targeted drugs of sodium channelopathies in the clinical treatment. In addition, a novel cysteine-rich secretory protein (CRBGP) has been isolated and purified from the buccal gland of the lampreys (Lampetra japonica), and it could inhibit the Na+ current of the hippocampus and dorsal root neurons for the first time. In the present study, the progress of the sodium channelopathies and the biological functions of voltage-gated sodium channel blockers are analyzed and summarized.
Animals ; Channelopathies ; physiopathology ; Hippocampus ; drug effects ; Neurons ; drug effects ; Neurotoxins ; pharmacology ; Venoms ; chemistry ; Voltage-Gated Sodium Channel Blockers ; pharmacology

This research was to study the regulation of intravenous administration of human umbilical cord blood mesenchymal stem cells (HUCBMSCs) on secretion of neural specific protein in rats after traumatic brain injury (TBI), and to explore its mechanisms promoting the recovery of neurological function. The TBI models of rats were established. We then injected HUCBMSCs, labelled by Brdu (5-bromo-2-deoxyuridine), into the TBI rats via the tail vein using modified Feeney free-falling method. The levels of neural biochemical indicators (serum S100β protein, NSE, LDH, CK) of rats were detected in shamed group, injury group and HUCBMSCs-transplanted group. And the morphological changes of brain tissue of rats in the three groups were observed by using HE staining under light microscope. During the whole experiment no immunosuppressant was used for the four groups. From the research, transplant-related death of the rats was not found in transplantation group. In the injury group, rises were found in contents of serum S100β protein, NSE, LDH, CK in the early stage after the rats were injured, which were much higher than those in shamed group at correspondent time point (P < 0.01). In HUCBMSCs-transplanted group, although these biochemistry indexes were found rising for a short period in the early stage, along with the time, these indexes were obviously lower than in those injury group (P < 0.05). Under light microscopy pathological changes of rats in HUCBMSCs-transplanted group were much slighter than those in injury group. It was well concluded that in the situation of no immuno-suppressants, the intravenous-injected HUCBMSCs could reduce the secretion of serum S100β protein, NSE, LDH, CK, promote the repair of tissue injury effectively, and promote the functional recovery of neurons.
Animals ; Biomarkers ; chemistry ; Brain ; pathology ; Brain Injuries ; therapy ; Cord Blood Stem Cell Transplantation ; Humans ; Mesenchymal Stem Cell Transplantation ; Neurons ; chemistry ; Rats