OBJECTIVETo investigate the effect of nerve growth factor (NGF) on lipopolysaccharide (LPS) injury and activation of nuclear factor-kappa B in PC12 cells.

METHODSIn order to set injury models, the PC12 cells were incubated in different concentration of LPS. Cells were cultured in the culture and were reduced by LPS, and then cells were treated by NGF of various concentrations. The cell viability was determined by methyl thiazolyl tetrazolium (MTT) assay, cellular morphology was observed under inverted microscope and fluorescence microscope, and the content of NF-kappaB was assessed by RT-PCR.

RESULTS(1) The viability of PC12 cell was decreased with concentration of LPS increasing. (2) The cellular morphology change showed that NGF had an ability to reduce LPS injury. (3) The result of RT-PCR showed that the content of NF-kappaB in LPS injury was more than the normal and treated cell, and the treated one was close to the normal one.

CONCLUSIONThe reports about NGF in brain cells repair after inflammatory are very small. And our study is about that NGF can protect the PC12 cell from LPS injury, and this mechanism possible bears on the activation of NF-kappaB.

Animals ; Lipopolysaccharides ; toxicity ; NF-kappa B ; metabolism ; Nerve Growth Factor ; pharmacology ; Neuroprotective Agents ; pharmacology ; PC12 Cells ; Rats

OBJECTIVETo explore the effect of 2,5-hexanedione (2,5-HD) on the levels of nerve growth factor (NGF) in sciatic nerve of rats and motor-neurons.

METHODA total of 50 Wistar rats were randomly designed into five groups and intoxicated with 400 mgxkg(-1)xd(-1) 2,5-HD for 0, 7, 14, 21, 28 d. Immunohistochemistry and real-time PCR were used to detect the levels of NGF and NGF mRNA. Motor neuron VSC4.1 cells were administrated with 0, 2.5, 5.0, 10.0, 20.0 mmol/L 2,5-HD for 24 h and 10.0 mmol/L 2,5-HD was chosen to intoxicated VSC4.1 cells for 0, 1, 3, 6, 12, 24, 48 h respectively. Immunofluorescence technique was selected to detect the levels of NGF.

RESULTSThe NGF level in sciatic nerve of rats administrated with 400 mgxkg(-1)xd(-1) 2,5-HD showed increase tendency at begin and then decrease after exposure. The NGF mRNA level in 14 d (2(-DeltaDeltaCt)= 3.46), 21 d (2(-DeltaDeltaCt)= 5.28) and 28 d (2(-DeltaDeltaCt)= 3.10) were higher than those in 0 d (2(-DeltaDeltaCt)= 1) and 7 d (2(-DeltaDeltaCt)= 0.78). In vitro tests of VSC4.1 cells showed that NGF levels in 5.0 mmol/L (43.24 +/- 7.52), 10.0 mmol/L (43.48 +/- 10.86) and 20.0 mmol/L (63.13 +/- 10.68) were higher than those in 0 mmol/L (16.32 +/- 4.20)(q values were 19.92, 19.72, 32.78, respectively, P < 0.01) and 2.5 mmol/L (19.78 +/- 2.66) (q values were 17.50, 17.42, 30.63, respectively, P < 0.01) in 24 h and the NGF level in 20.0 mmol/L was higher than those in 5.0 mmol/L (q = 13.04, P < 0.01) and 10.0 mmol/L (q = 11.71, P < 0.01). The NGF levels of VSC4.1 cells with 10.0 mmol/L 2,5-HD in 6 h (18.66 +/- 2.89), 12 h (23.14 +/- 6.08), 24 h (27.66 +/- 6.11) and 48 h (17.25 +/- 3.05) were increased compared with that in 0 h (10.18 +/- 1.81) (q values were 9.64, 15.74, 21.76, 8.50, respectively, P < 0.01), 1 h (9.31 +/- 1.28) (q values were 10.28, 16.17, 21.95, 9.20, respectively, P < 0.01) and 3 h (10.44 +/- 2.13) (q values were 9.25, 15.24, 21.17, 8.10, respectively, P < 0.01), and NGF levels in 12 h and 24 h increased compared with those in 6 h (q values were 5.24, 10.77, respectively, P < 0.01) and 48 h (q values were 7.31, 13.26, respectively, P < 0.01).

CONCLUSION2,5-HD could increase NGF levels in sciatic nerve of rats and motor-neurons, and the dose or time dependent effects were observed in this study.

Animals ; Cell Line ; Hexanones ; toxicity ; Male ; Motor Neurons ; drug effects ; metabolism ; Nerve Growth Factor ; metabolism ; Rats ; Rats, Wistar ; Sciatic Nerve ; drug effects ; metabolism

OBJECTIVETo investigate the effects of high-affinity tyrosine kinase receptors TrkB, TrkC and the low-affinity neurotrophin receptor p75 in spiral ganglion cell (SGC) of cisplatin-induced ototoxicity.

METHODSThe 50 adult Wistar rats were divided randomly into 5 groups received intraperitoneal injection of cisplatin with vary dose. Control group was received equivalent volumes of saline. The group received 1 day intraperitoneal injection was cisplatin treated at a dose of 5 mg/kg and killed at next day. The group received 3 days was cisplatin treated for 3 days at same dose daily and then killed at next day. The group A received 5 days was cisplatin treated for 5 days and killed at next day. The group B received 5 days was cisplatin treated for 5 days and then were sacrificed after 7 days. The change of mRNA level of neurotrophin receptors in cochlear tissue were examined by RT-PCR. The expressing pattern of TrkB, TrkC, P75 in damaged cochlea were study by immunochemistry using antibodies against TrkB, TrkC, P75 protein.

RESULTSThe research data showed the expression of Trk B, Trk C, p75 exhibited in SGC was dynamic along with the administration lasting. The mRNA and protein level of Trk B (x(-) +/- s) at day 1 and 3 after cisplatin treatment were 0.76 +/- 0.06, 88.78 +/- 4.28, 0.82 +/- 0.09 and 91.64 +/- 4.06, with significant difference among those and other groups (P < 0.05). The mRNA and protein level of TrkC at day 1 after cisplatin treatment were 0.80 +/- 0.06 and 89.66 +/- 2.76, with significant difference among that and other groups (P < 0.05). The mRNA and protein level of p75 at the control group and cisplatin treated groups were 0.64 +/- 0.04, 55.16 +/- 3.10, 0.77 +/- 0.04, 78.46 +/- 3.86, 1.01 +/- 0.09, 105.02 +/- 6.61, 1.18 +/- 0.09, 111.10 +/- 6.08, 0.51 +/- 0.04 and 42.74 +/- 5.20, with significant difference among the control group and cisplatin treated groups (P < 0.05).

CONCLUSIONSThe expression of Trk B increased to peak at day 1 - 3 after cisplatin treatment and decreased at day 5 early and following weeks. The expression of Trk C went up to peak at day 1 after cisplatin treatment and went down during subsequently time. P75 kept a trend of continuance increased during the drug treatment and decrease at drug stopped. The expression of Trk B, Trk C and P75 may be involved in cochlear insult with cisplatin-induced. Trk B and Trk C may play an important role in the reparative process of cochlear, especially at early stage of the damage. P75 could promote SGC apoptosis in cisplatin-induced neurotoxicity.

Animals ; Cisplatin ; toxicity ; Male ; Rats ; Rats, Wistar ; Receptor, Nerve Growth Factor ; metabolism ; Receptor, trkB ; metabolism ; Receptor, trkC ; metabolism ; Spiral Ganglion ; drug effects ; metabolism

AIMTo investigate the preventive effects of the cerebro cellular growth peptide (CCGP) on gentamycin-induced inner ear damage in guinea pigs, and to clarify its mechanism.

METHODSThe hypoacusis severity and enzymatic activity in the cochlear hair cells were examined by brainstem auditory evoked potential (BAEP) and histochemistry, respectively. The damaged hair cells was counted in three groups.

RESULTSCCGP reduced the elevated BAEP reaction thresholds. It protected activities of mitochondrial succinate dehydrogenase and lysosome acid phosphatase in the cochlear hair cells. The number of damaged hair cells in the CCGP group was less than that in the gentamycin (GM) group.

CONCLUSIONCCGP can reduce GM ototoxicity. The mechanism may be associated with the protective activity of mitochondrial enzyme, the maintenance of lysosome intactness, energy metabolism of the cochlear hair cells, and reduction of autolysis of hair cells induced by hydrolase over flowing from lysosome.

Animals ; Evoked Potentials, Auditory, Brain Stem ; Female ; Gentamicins ; toxicity ; Guinea Pigs ; Hair Cells, Auditory ; drug effects ; physiology ; Male ; Nerve Growth Factor ; pharmacology

OBJECTIVESTo study the effect of lead acetate on the expression of nerve growth factor (NGF) protein in rat brain and the regulation of thyroid hormone.

METHODSLead acetate was given to SD rats intraperitoneally ip. at the dosage of 25, 50 and 100 mg/kg respectively. 6-n-propyl-2-thiouracil (PTU) was used to make a hypothyroid model and then lead acetate was given at the dosage of 50 mg/kg body weight through i.p. The NGF protein expression in rat brain was observed by immunohistochemistry Triiodothyronine (T3), thyroxin (T4), TSH in serum and T3, T4 in brain tissue were determined by radio immunoassays (RIAs).

RESULTSThe average gray value of NGF protein in cerebral cortex of 50 mg, 100 mg treated groups (180.49 +/- 10.33, 169.72 +/- 19.75, respectively) were lower than the control (200.75 +/- 3.27, P<0.01). The area density of NGF protein in hippocampus of three treated groups (0.08 +/- 0.14, 0.12 +/- 0.02, 0.13 +/- 0.04, respectively) were significantly different from the control (0.025 +/- 0.015, P<0.05). The area density and the average gray value of NGF protein in lead acetate treated hypothyroid rat brain were of no significant changes. The levels of serum T3 in three treated groups [(0.68 +/- 0.02), (0.57 +/- 0.04), (0.54 +/- 0.02) microg/L respectively] and T4 [(28.30 +/- 1.83), (27.35 +/- 2.55), (24.00 +/- 3.01) microg/L] in serum were significantly lower while TSH [(6.34 +/- 1.13), (7.74 +/- 0.79), (9.16 +/- 0.77) IU] higher than those in the control [T3 (0.97 +/- 0.14) microg/L, T4 (54.50 +/- 3.70) microg/L and TSH (4.62 +/- 2.16) IU], and there was a good dose-response relationship. The levels of T3 in cerebral cortex of three treated groups [(13.26 +/- 0.81), (11.49 +/- 0.10), (10.42 +/- 1.19) pg/mg pro respectively] and T4 [(0.50 +/- 0.03), (0.49 +/- 0.13), (0.42 +/- 0.01) ng/mg pro] were significantly lower than those in control [(20.85 +/- 11.01) pg/mg pro, (0.76 +/- 0.14) ng/mg pro, P<0.05, P<0.01].

CONCLUSIONLead could increase the NGF protein expression in rat brain, which may be regulated by thyroid hormone.

Animals ; Brain Chemistry ; drug effects ; Immunohistochemistry ; Male ; Nerve Growth Factor ; analysis ; Organometallic Compounds ; toxicity ; Rats ; Rats, Sprague-Dawley ; Thyroid Hormones ; analysis ; blood ; physiology

This study was performed to evaluate the effects of nerve growth factor (NGF) upon angiogenesis in the rat cornea, to examine its possible application as an alternative angiogenic inducer and to provide basic data for further studies. Angiogenesis was induced by cornea micropocket assay, as previously described. Eight of thirty two eyes of Sprague-Dawley rats were randomly assigned to one of four groups, namely, a non-NGF group (Group 0), a 0.5 ng of NGF group (Group 0.5), a 1.0 ng of NGF group (Group 1.0) and a 5.0 ng of NGF group (Group 5.0). Pellets made of poly-2-hydroxylethylmethacrylate and sucralfate were implanted into the corneal stroma no closer than 1 mm from the limbus. After the implantation, the number of new vessels, vessel length and circumferential neovascularization were examined daily under the surgical microscope over a period of 7 days. The area of neovascularization was determined using a mathematical formula. Although new vessels in Group 0 and Group 0.5 were first observed at day 5, those of Groups 1.0 and 5.0 were first noted on days 4 and 3, respectively. However, the growth rates of new vessels in Groups 1.0 and 5.0 were higher than those of Groups 0 and 0.5 with the passage of time. The number, length, circumferential neovascularization and areas covered by the vessels in Groups 1.0 and 5.0 were significantly more than in Group 0 and Group 0.5 (p<0.05). This study showed that NGF had a dose-dependent angiogenic effects on the rat cornea and that the minimal effective dose of NGF was 1.0 ng per cornea. Also, it showed that NGF would be useful in angiogenic studies as an alternative angiogenic inducer.
Angiogenesis Inducing Agents/*toxicity ; Animals ; Cornea/blood supply/*drug effects ; Corneal Neovascularization/*chemically induced ; Dose-Response Relationship, Drug ; Female ; Male ; Nerve Growth Factor/*toxicity ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the effects of lead acetate on the expression of brain-derived neurotropic factor (BDNF) and its receptor P75NTR in rat brain.

METHODSLead acetate was given to SD rats by intraperitoneal injection (ip) for 5 days at the dosage of 25, 50 and 100mg/kg body weight respectively. The contents of lead in serum, cerebral cortex and hippocampus were measured by atomic absorption spectrophotochemistry. The levels of BDNF mRNA and protein expression in cerebral cortex and hippocampus were observed by RT-PCR and immunohistochemistry, respectively. The levels of P75NTR protein expression in rat brain were measured by immunohistochemistry.

RESULTSCompared with the control, the contents of lead were significantly increased in serum, cerebral cortex and hippocampus in the treatment groups respectively (P < 0.01, P < 0.05). The BDNF mRNA expression in the cerebral cortex (0.52 +/- 0.05, 0.33 +/- 0.03) and hippocampus (0.77 +/- 0.10, 0.92 +/- 0.08) of 50, 100 mg/kg treated groups was significantly higher than that of the control group (0.52 +/- 0.05, 0.33 +/- 0.03), respectively (P < 0.05). The results of immunohistochemistry showed that the area density of BDNF protein in cerebral cortex of every treatment group (0.040 +/- 0.027, 0.048 +/- 0.027, 0.086 +/- 0.040) was significantly increased whereas the average gray value (187.11 +/- 11.15, 180.53 +/- 5.82, 180.15 +/- 8.01) was significantly lower than that of the control (0.026 +/- 0.005, 204.98 +/- 3.45) (P < 0.05, P < 0.01). The area density of BDNF protein in hippocampus of every treatment group was 0.040 +/- 0.027, 0.048 +/- 0.027, 0.086 +/- 0.040, respectively, which was significantly increased compared with the control (0.045 +/- 0.019, P < 0.05). The average gray value of BDNF protein in hippocampus (181.03 +/- 5.16, 171.25 +/- 12.65) of 50, 100 mg/kg were significantly lower than that of the control (198.98 +/- 6.40, P < 0.01). There was no positive expression of P75NTR protein in the control and 25 mg/kg body weight groups. The positive expression of P75NTR protein was detected in 50 and 100 mg/kg body weight groups.

CONCLUSIONLead can increase the BDNF and P75NTR expression in rat brain which might play an important role in the neural damage and repair.

Animals ; Brain ; drug effects ; metabolism ; Brain-Derived Neurotrophic Factor ; biosynthesis ; Dose-Response Relationship, Drug ; Female ; Immunohistochemistry ; Male ; Organometallic Compounds ; toxicity ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptor, Nerve Growth Factor ; biosynthesis ; Reverse Transcriptase Polymerase Chain Reaction

Due to the therapeutic potential of gene therapy for neuronal injury, many studies of neurotrophic factors, vectors, and animal models have been performed. The presumed dog beta-nerve growth factor (pdbeta-NGF) was generated and cloned and its expression was confirmed in CHO cells. The recombinant pdbeta-NGF protein reacted with a human beta-NGF antibody and showed bioactivity in PC12 cells. The pdbeta-NGF was shown to have similar bioactivity to the dog beta-NGF. The recombinant pdbeta-NGF plasmid was administrated into the intrathecal space in the gene therapy group. Twenty-four hours after the vector inoculation, the gene therapy group and the positive control group were intoxicated with excess pyridoxine for seven days. Each morning throughout the test period, the dogs' body weight was taken and postural reaction assessments were made. Electrophysiological recordings were performed twice, once before the experiment and once after the test period. After the experimental period, histological analysis was performed. Dogs in the gene therapy group had no weight change and were normal in postural reaction assessments. Electrophysiological recordings were also normal for the gene therapy group. Histological analysis showed that neither the axons nor the myelin of the dorsal funiculus of L(4) were severely damaged in the gene therapy group. In addition, the dorsal root ganglia of L(4) and the peripheral nerves (sciatic nerve) did not experience severe degenerative changes in the gene therapy group. This study is the first to show the protective effect of NGF gene therapy in a dog model.
Amino Acid Sequence ; Animals ; Base Sequence ; CHO Cells ; Central Nervous System Diseases/chemically induced/therapy/*veterinary ; Cloning, Molecular ; Cricetinae ; Cricetulus ; Cytomegalovirus ; Dog Diseases/*chemically induced/therapy ; Dogs ; Female ; Gene Therapy/*veterinary ; Genetic Vectors ; Male ; Molecular Sequence Data ; Nerve Growth Factor/genetics/*metabolism/*therapeutic use ; Pyridoxine/*toxicity