Animals ; Epilepsy ; immunology ; metabolism ; Hippocampus ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; gamma-Aminobutyric Acid ; immunology ; metabolism

AIM AND METHODSThe effect of intrahippocampal microinjection of noradrenaline (NA) and its receptors antagonists and agonists on cellular immune functions were investigated in normal and adrenalectomy rat by determine the proliferative activity of Con A-stimulated splenic lymphocytes in MTT method and natural killer (NK) cell activity.

RESULTS(1) In normal group, the proliferative activity of Con A-Stimulated splenic lymphocytes were inhibited and the activity of NK cell were reduced with microinjection NA and beta1-, beta2-adrenergic receptor agonists Dobutamine (Dob, 4 microl, 6.0 x 10(-3) moL/L), Metaproterenol (Met, 4 microl, 8.0 x 10(-3) mol/L), compared with their intensity of effect, NA > Met > Dob; the immunosuppression effect induced by NA was partly hindered by alpha- and beta-receptor antagonists, phentolamine (Phen, 2 microl, 1.6 x 10(-2) mol/L) and propranolol (Prop, 2 microl, 1.6 x 10(-3) mol/L), and the action of Prop was more evident. (2) In adrenalectomy group, immunosuppression effect induced by NA was unconspicuous.

CONCLUSIONThe results suggested that NA in hippocampus could inhibit distinctly cellular immune functions, which was predominantly mediated by beta2- adrenergic receptor with a minor contribution of beta1- and alpha- adrenergic receptors. Moreover, keeping intact construction and function of adrenal gland have an important role in the effect of NA on cellular immune function.

Adrenergic Agonists ; pharmacology ; Animals ; Hippocampus ; drug effects ; Immunity, Cellular ; drug effects ; Killer Cells, Natural ; immunology ; Lymphocytes ; immunology ; Microinjections ; Norepinephrine ; pharmacology ; Rats ; Rats, Wistar ; Spleen ; cytology ; immunology

OBJECTIVETo observe the effects of transcutaneous electrical acupoint stimulation (TEAS) combined with general anesthesia or controlled hypotension on hippocampal neuronal damage and the inflammatory response in peripheral circulation and central nervous system (CNS) after surgery, and to investigate its brain protection mechanism.

METHODSEighteen healthy male beagles aged 6 - 8 months were randomly divided into a general anesthesia group (group G), a controlled hypotension group (group C) and a compound anesthesia acupuncture group (group A), 6 cases in each group. Dogs in group G was anesthetized by isoflurane inhalation, and group C was combined with intravenous infusion of sodium nitroprusside based on isoflurane inhalation to induce hypotension, and followed surgery after achieving the target blood pressure, and group A was combined with TEAS at "Quchi" (LI 11), "Hegu" (LI 4) "Zu sanli" (ST 36) and "Sanyinjiao" (SP 6) based on controlled hypotension, and then brain tissue was taken out on the 72 h after mean arterial pressure (MAP) was returned to baseline levels. The concentration of IL-1beta,TNF-alpha in serum at different time points were detected by ELISA. The expression of IL-1beta, TNF-alpha, Bcl-2, Bax and cleaved caspase-3 were measured by immunohistochemistry, and the apoptosis of hippocampus were detected by TUNEL.

RESULTS(1) At different time points, the concentration of TNFalpha showed the trend of increase first and then decrease, while IL-1beta concentration represented a trend of decrease first and then increase in both group C and group A, but there were no significant differences in cytokine expression between the two groups (all P > 0.05). (2) The ratio of positive cells of IL-1beta, TNF-alpha and caspase-3 in CA1 and CA3 of hippocampus in both group C and A were higher than those in group G (all P < 0.01), and cytokines expression in group A were lower than those in group C (all P < 0.01), and caspase-3 in CA1 in group A was lower than that in group C (P < 0.01). The ratio of Bcl-2/Bax in both group C and A were lower than that in group G (all P < 0.01), and that in group A was higher than that in group C (P < 0.01 in CA1, P < 0.05 in CA3). (3) The apoptosis index (AI) of hippocampal neurons in both group C and A was significantly higher than that in group G (P < 0.01), while AI in CA1 in group A was lower than that in group C (P < 0.01).

CONCLUSIONThe TEAS can regulate the expression of inflammatory factor in hippocampus in animals undergoing general anesthesia or con trolled hypotension surgery, further improving Bcl-2/Bax ratio, inhibiting the expression of caspase-3 and reducing neuron apoptosis in hippocampus so as to play a neuroprotection.

Acupuncture Analgesia ; Acupuncture Points ; Anesthesia, General ; Animals ; Apoptosis ; Dogs ; Hippocampus ; cytology ; immunology ; surgery ; Humans ; Hypotension, Controlled ; Inflammation ; genetics ; immunology ; physiopathology ; therapy ; Interleukin-1beta ; genetics ; immunology ; Male ; Neurons ; cytology ; immunology ; Transcutaneous Electric Nerve Stimulation ; Tumor Necrosis Factor-alpha ; genetics ; immunology

The hippocampus is a central area of the memory-related neural system. Combined immunohistochemistry against choline acetyl transferase and retrograde transneuronal labelling of the pseudorabies virus were used to identify cholinergic neurons in the central nervous system projecting to the hippocampal formation of the rat. Five to ten microL of Bartha strain of pseudorabies virus were injected into the dentate gyrus, CA1 and CA3 of the hippocampus of 20 Sprague Dawley rats using stereotaxic instrument. Forty eight to 96 hr after the injection, the brains were removed and the tissue sections were processed for double immunofluorescence procedure using polyclonal antibodies against pseudorabies virus or choline acetyl transferase. The double labelled neurons were distributed at several different nuclei and the labelling patterns of three different areas of the hippocampus were similar. These data suggests that the cholinergic innervation to the hippocampus were distributed in a transsynaptic manner throughout the whole brain area.
Animal ; Antibodies ; Choline O-Acetyltransferase/*analysis/immunology ; Cholinergic Fibers/*enzymology ; Herpesvirus 1, Suid/immunology ; Hippocampus/*cytology ; Immunohistochemistry ; Microinjections ; Neural Pathways ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the effects of immunization with the fusion protein CAC (a product of prokaryotic expression of recombinant HBcAg and β-amyloid peptide fusion gene) against the toxicity induced by intrahippocampal injection of aggregated β-amyloid peptide (Aβ) in rats.

METHODSSD rats were immunized intraperitoneally with the fusion protein CAC, and the titer of anti-Aβ antibody was evaluated by ELISA. When the titers of the anti-Aβ antibody reached 1:3 000, aggregated Aβ was injected into the CA1 region of the rat hippocampus. Two weeks after Aβ injection, the rats underwent morris water maze test before sacrificed to prepare the brain slices with Congo red and haematoxylin staining.

RESULTSThe titer of anti-Aβ antibody reached 1:3 000 after 5 immunizations with the fusion protein. After Aβ injection, the saline-immunized rats showed a reduced cognitive behavior in the Morris water maze test compared to the CAC-immunized rats. In the saline-immunized rats, the neurons around the site of Aβ injection exhibited obvious cell damages with Aβ deposits and glial infiltration, whereas in CAC-immunized rats, Aβ deposits were significantly reduced or even absent.

CONCLUSIONImmunization with the fusion protein CAC can inhibit the toxicity induced by intrahippocampal aggregated Aβ injection.

Amyloid beta-Peptides ; administration & dosage ; biosynthesis ; genetics ; immunology ; Animals ; Antibodies ; blood ; Hepatitis B Core Antigens ; biosynthesis ; genetics ; Hippocampus ; metabolism ; Immunization ; Injections ; Male ; Peptide Fragments ; administration & dosage ; immunology ; Rats ; Rats, Sprague-Dawley ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology

OBJECTIVETo study the effects of electroacupuncture (EA) on the changes of behavior after ketamine anesthesia, and changes of serum antibodies against beta-amyloid (Abeta) and Abeta protein in the hippocampus of aged rats, thus exploring the effects of EA on the cognitive dysfunction.

METHODSThirty 14-month old SD rats were randomly divided into 3 groups, i. e. , the control group (Group A), the ketamine anesthesia group (Group B), and the EA+ketamine anesthesia group (Group C), 10 in each group. 50 mg/kg katemine was intraperitoneally injected to rats in Group B and Group C, once daily for 7 successive days. EA was performed to rats in Group C from the 1st day of the experiment after rats awoke completely from anesthesia, twice daily for 7 successive days. Changes of the ratio of the swim time in the original platform quadrant to the total swim time and the escape latency phase were observed by Morris water maze. The peripheral blood was withdrawn by the end of the experiment. Serum anti-Abeta antibody contents were detected using enzyme-linked immunosorbent assay (ELISA). The expressions of Abeta in the hippocampus were detected using Westen blot.

RESULTSLong-term application of ketamine could lower aged rats' cognitive function. In the navigation test, the escape latency phase of rats in Group B was significantly prolonged ( P < 0.01) . On the 7th day of the experiment, the serum level of anti-Abeta antibodies was lower in Group B than in Group A (P < 0.05), while the serum level of anti-Abeta antibodies was significantly higher in Group C than in Group B (P < 0.01). On the 7th day of the experiment, the expression of Abeta in the hippocampus was higher in Group B than in Group A (P < 0.05).

CONCLUSIONEA could increase the contents of anti-Abeta antibodies in aged rats with ketamine anesthesia, decrease the expression of Abeta in the hippocampus, alleviate the deposition of Abeta, thus improving rats' cognitive dysfunction.

Amyloid beta-Peptides ; immunology ; Anesthesia ; adverse effects ; Animals ; Antibodies ; blood ; Cognitive Dysfunction ; therapy ; Electroacupuncture ; Female ; Hippocampus ; metabolism ; Ketamine ; adverse effects ; Male ; Maze Learning ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the regulating effect of Ginkgo biloba extract 50 (GBE50) on pre-inflammatory factors interleukin-1 beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha) and anti-inflammatory factors interleukin-4 (IL-4), interleukin-10 (IL-10) of hippocampus in senile rats, in order to explore the protective mechanism of GBE50 on central nervous system of senile animals.

METHODSD rats were randomly divided into four groups: the normal group, the model group, the GBE50 group and the EGB761 group. Rats were intraperitoneally injected with 100 mg x kg(-1) D-galactose every day for 42 days to establish the senile rat model. At the 21st day, the GBE50 group and the EGB761 group were orally administered with 60 mg x kg(-1) for 21 days. IL-1beta mRNA and TNF-alpha mRNA expressions were detected by real-time fluorescence quantitative PCR assay, IL-1beta and TNF-alpha protein expressions were detected by immunohistochemistry, IL-4 and IL-10 protein contents were detected by ELISA.

RESULTD-galactose caused imbalance between pre-inflammatory factors and anti-inflammatory factors of hippocampus in senile rats, GBE50 and EGB761 reduced IL-1beta mRNA expression (P < 0.05) and TNF-alpha and IL-1beta protein level (P < 0.01) and up-regulated IL-10 protein content (P < 0.01, P < 0.05).

CONCLUSIONThe mechanism of GBE50 in protecting central nervous system is probably related to its effect in mitigating inflammatory of central nervous system.

Animals ; Ginkgo biloba ; Hippocampus ; drug effects ; immunology ; Interleukin-1beta ; analysis ; Male ; Plant Extracts ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; analysis

OBJECTIVETo study the effects of immunoglobulin on the neuronal expression of IL-1beta and IL-1ra and the neuronal death at hippocampus in rats with convulsion induced by pentylenetetrazol.

METHODSThe epilepsy model was established by injecting intraperitoneally pentylenetetrazol (PTZ) into Wistar rats. Forty-five rats were randomly divided into three groups, normal control group, PTZ plus intravenous immunoglobulin (PTZ-IVIG); PTZ plus normal saline (PTZ-NS). Neuronal death was assessed by light microscopy with the hematoxylin-eosin (HE) staining and with in situ terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL). IL-1beta and IL-1ra expressions were examined by histochemistry.

RESULTSThe ratio of IL-1beta/IL-1ra at hippocampal CA(1) region in PTZ-IVIG group (0.5 +/- 0.1) was significantly lower than that in PTZ-NS group (1.9 +/- 0.5, t = 12.9, P < 0.05). Apoptotic cell numbers at the hippocampal CA(1) region were significantly decreased in the PTZ-IVIG group, compared to PTZ-NS group (t = 27.1, P < 0.05). The numbers of positive cells were 16.4 +/- 3.3/1000 microm(2) in the former and 41.7 +/- 3.5/1000 microm(2) in the latter. Necrotic cell numbers at the hippocampal CA(1) region were significantly decreased in the PTZ-IVIG group (19.0 +/- 2.6/1000 microm(2)), compared to PTZ-NS group (42.3 +/- 4.9/1000 microm(2), t = 20.9, P < 0.05).

CONCLUSIONImmunoglobulin could inhibit neuronal death induced by convulsion and its possible mechanism might be the regulation of IL-1 system in neurons.

Animals ; Apoptosis ; Hippocampus ; drug effects ; immunology ; metabolism ; Immunoglobulins, Intravenous ; pharmacology ; Interleukin 1 Receptor Antagonist Protein ; metabolism ; Interleukin-1beta ; metabolism ; Neurons ; drug effects ; Pentylenetetrazole ; adverse effects ; Rats ; Rats, Wistar ; Seizures ; chemically induced ; immunology ; metabolism

This study examined whether amifostine (WR-2721) could attenuate memory impairment and suppress hippocampal neurogenesis in adult mice with the relatively low-dose exposure of acute radiation syndrome (ARS). These were assessed using object recognition memory test, the terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling assay, and immunohistochemical markers of neurogenesis [Ki-67 and doublecortin (DCX)]. Amifostine treatment (214 mg/kg, i.p.) prior to irradiation significantly attenuated the recognition memory defect in ARS, and markedly blocked the apoptotic death and decrease of Ki-67- and DCX-positive cells in ARS. Therefore, amifostine may attenuate recognition memory defect in a relatively low-dose exposure of ARS in adult mice, possibly by inhibiting a detrimental effect of irradiation on hippocampal neurogenesis.
Acute Radiation Syndrome/drug therapy/*immunology/psychology ; Amifostine/*pharmacology/therapeutic use ; Animals ; Apoptosis/immunology ; Gamma Rays/*adverse effects ; Hippocampus/immunology ; Immunohistochemistry ; In Situ Nick-End Labeling ; Male ; Memory/*radiation effects ; Mice ; Mice, Inbred ICR ; Neurogenesis/immunology ; Radiation-Protective Agents/*pharmacology/therapeutic use

OBJECTIVE: To explore the application of immunofluorescence and sandwich ELISA with double-antibodies in detection of human rabies. METHODS: The cerebrum, cerebellum, brainstem, and hippocampus of four patients died of rabies identified by clinical diagnosis were collected and kept in freezer at -70 degrees C or in formaldehyde solution separately. The rat brain tissue infected by CVS strain of rabies virus was used as a positive control and the brain tissue of a patient died of acute pancreatitis was used as a negative control. RESULTS: Rabies virus was detected in the tissues kept in freezer at -70 degrees C and the positive control but was not detected in the tissues kept in formaldehyde solution and the negative control. CONCLUSION Immunofluorescence and Sandwich ELISA with double-antibodies could be used in detection of human rabies. The samples should be kept in deep frozen temperature condition instead of in formaldehyde solution.
Animals ; Antibodies, Viral/analysis* ; Brain/virology* ; Enzyme-Linked Immunosorbent Assay/methods* ; Fluorescent Antibody Technique/methods* ; Hippocampus/virology* ; Humans ; Rabies/virology* ; Rabies virus/immunology* ; Rats ; Reagent Kits, Diagnostic ; Sensitivity and Specificity ; Tissue Preservation/methods*