1.Effect of Sodium Salicylate on Calcium Currents and Exocytosis in Cochlear Inner Hair Cells: Implications for Tinnitus Generation.
Ting FAN ; Meng-Ya XIANG ; Ruo-Qiao ZHOU ; Wen LI ; Li-Qin WANG ; Peng-Fei GUAN ; Geng-Lin LI ; Yun-Feng WANG ; Jian LI
Neuroscience Bulletin 2022;38(1):69-80
Sodium salicylate is an anti-inflammatory medication with a side-effect of tinnitus. Here, we used mouse cochlear cultures to explore the effects of salicylate treatment on cochlear inner hair cells (IHCs). We found that IHCs showed significant damage after exposure to a high concentration of salicylate. Whole-cell patch clamp recordings showed that 1-5 mmol/L salicylate did not affect the exocytosis of IHCs, indicating that IHCs are not involved in tinnitus generation by enhancing their neuronal input. Instead, salicylate induced a larger peak amplitude, a more negative half-activation voltage, and a steeper slope factor of Ca2+ current. Using noise analysis of Ca2+ tail currents and qRT-PCR, we further found that salicylate increased the number of Ca2+ channels along with CaV1.3 expression. All these changes could act synergistically to enhance the Ca2+ influx into IHCs. Inhibition of intracellular Ca2+ overload significantly attenuated IHC death after 10 mmol/L salicylate treatment. These results implicate a cellular mechanism for tinnitus generation in the peripheral auditory system.
Animals
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Calcium
;
Exocytosis
;
Hair Cells, Auditory, Inner
;
Mice
;
Sodium Salicylate/pharmacology*
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Tinnitus/chemically induced*
2.Inhibition of salicylate on voltage-gated sodium channels in rat inferior colliculus neurons.
Yanxing LIU ; Xuepei LI ; Hailin ZHANG ; Yongli WANG ; Hong LU ; Xiang QI ; Huijun ZHANG
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2007;21(19):890-894
OBJECTIVE:
To investigate the mechanism of the tinnitus inducer, sodium salicylate, on voltage-gated sodium channels.
METHOD:
The effects of salicylate on voltage-gated sodium channels in freshly dissociated inferior colliculus neurons of rats were studied, using the whole-cell voltage clamp method.
RESULT:
Salicylate blocked sodium current (INa) in concentration-dependent manner (0.1-10 mmol/L). The IC50 value for the blocking action of salicylate was 1.43 mmol/L. Salicylate did not affect the conductance-voltage curve and the steady-state activation curve of INa. The steady-state INa inactivation curve of INa was shifted by about 9 mV in the hyperpolarizing direction. In addition, salicylate delayed the sodium channel recovery from INa inactivation by increasing the slow time constant.
CONCLUSION
Our results suggest that salicylate causes a concentration-dependent blockade of INa and shifts the INa inactivation curve to more hyperpolarized potentials, which could be related to the mechanism of salicylate-induced tinnitus.
Animals
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Inferior Colliculi
;
cytology
;
Male
;
Neurons
;
drug effects
;
metabolism
;
Patch-Clamp Techniques
;
Rats
;
Rats, Wistar
;
Sodium Channels
;
drug effects
;
metabolism
;
Sodium Salicylate
;
pharmacology
3.Differentially expressed gene profiles of cochlea in rats induced by acute and chronic sodium salicylate injection.
Zhi-Wu HUANG ; Ping CHEN ; Ling MEI ; Qi-Jun FAN ; Bo-Kui XIAO ; Zhan-Yuan WU
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2005;40(1):27-32
OBJECTIVETo study the mechanism of electrophysiologic changes caused by different type of sodium salicylate injection.
METHODSDecapitated three group rats ( acute injected, chronic injected and normal rats ) separately, dissected the temporal bones to collect cochlea, and the otic capsules were removed. Then the cochlear materials from each groups were pooled and homogenized respectively, extracted the total RNA, obtained cDNA from purified total RNA by reversed transcription, cDNA were transcripted to cRNA probes in vitro. Hybridized the cRNA probes with tester chip to evaluate the quality of probes, if good, hybridized the probes with real chip. Obtained three gene expression profiles of different groups of cochlea Analyzed the differentially expressed genes among three groups by SOM. Analogized the SOM result to electrophysiologic changes. Then analyzed the genes in clusters of analog results by Gene Ontology. Then the genes in clusters of analog results were analyzed by Gene Ontology. Hsp27 was chosen to validate the result of gene chip using real time quantitative reverse transcription PCR ( RTQ RT-PCR).
RESULTSThe probes was good, and the chip hybridization results was credible. We obtained 6 clusters genes by SOM analysis, in which we choose cluster 3 and cluster 4 as candidate cluster. There were 46 genes in cluster 3 and 30 genes in cluster 4 employing GO analysis, which involved in cell communication, cell motility, metabolism, immune response and nerve ensheathment, et al. The result of RTQ RT-PCR showed high concordance with that of gene chip.
CONCLUSIONIt's a new method to study the mechanism of electrophysiologic changes caused by sodium salicylate by gene chip and SOM analysis.
Animals ; Cochlea ; drug effects ; metabolism ; Gene Expression Profiling ; Injections ; Oligonucleotide Array Sequence Analysis ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar ; Sodium Salicylate ; administration & dosage ; pharmacology
4.Effect of sodium salicylate on the auditory brain stem response threshold and expression of glutamic acid decarboxylase in spiral ganglion of juvenile and adult guinea pigs.
Shi-Hua YIN ; An-Zhou TANG ; Song-Hua TAN ; Ping CHEN ; Li-Hong XIE ; Yi REN
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2008;43(5):364-368
OBJECTIVETo study the differences of regulation of sodium salicylate on the auditory brain stem response (ABR) threshold and expression of glutamic acid decarboxylase (GAD) protein in spiral ganglion of juvenile and adult guinea pigs.
METHODSFourty juvenile guinea pigs which were born just four days and fourty adult guinea pigs which were born thirty days were selected. They were divided four groups (group A; group B; group C; group D). ABR threshold was detected before administration, after administration for 15 days and after administration stopped for 30 days. The protein expression of GAD were measured after administration for 15 days and after administration stopped for 30 days by the method of immunohistochemistry.
RESULTSABR threshold of juvenile sodium salicylate groups (group C) was increased remarkably than that of before administration and the control after administration for 15 days (P < 0.001). ABR threshold of group C was returned to the level of that of before administration and after administration stopped for 30 days. ABR threshold of adult sodium salicylate groups (group D) was increased remarkably than that of before administration and the control after administration for 15 days (P < 0.001). ABR threshold of group D was kept the high level after administration stopped for 30 days. The protein expression of GAD of sodium salicylate groups (group C and D) was decreased than that of the control after administration for 15 days. The protein expression of group C was more visible regression than that of group D (t = 4.7, P < 0.001). The protein expression of group C was returned the level of before administration after administration stopped for 30 days, but the protein expression of group D was kept the high level.
CONCLUSIONSThe results suggest that sodium salicylate can regulate differently ABR threshold and expression of GAD protein in spiral ganglion of juvenile and adult guinea pigs. The effects of sodium salicylate on ABR threshold and expression of GAD protein in spiral ganglion of juvenile pigs are more noticeable than that of adult guinea pigs, but these changes are easier to return the normal than that of adult guinea pigs.
Animals ; Auditory Threshold ; drug effects ; Evoked Potentials, Auditory, Brain Stem ; drug effects ; physiology ; Glutamate Decarboxylase ; metabolism ; Guinea Pigs ; Sodium Salicylate ; pharmacology ; Spiral Ganglion ; drug effects ; enzymology
5.The effect of sodium salicylate on the expression of GABAa receptor subunits in cochlear spiral ganglion neurons.
Chen YAO ; Zheng CAI ; Renjun WANG ; Huiying CHEN ; Zhihui HUANG ; Jixin QIN ; Jiping SU
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2015;29(11):1024-1029
OBJECTIVE:
To investigate mRNA expression of GABAa receptor(GABAaR) subunits in the rat cochlear spiral ganglion neurons (SGN) and explore the effect of sodium salicylate (SS) on the expression of GABAaR subunits.
METHOD:
The realtime fluorescent quantitative PCR (FQ-PCR) was used to detect mRNA expression of twelve GABAaR subunits in the newborn rat SGN and then investigate mRNA expression of GABAaR subunits after treatment with 5 mmol/L SS for 15 min, 30 min, 1 h, 3 h and 6 h in the primary culture SGN.
RESULT:
(1) GABAaR subunits of α1-6, β1-3, and γ1-3 were detected in the SGN, and the expression of GABAaR subunits was lower than those in the cerebral cortex. In the subunit α family of GABAaR, the expression rank was α2>α3/α5>α4>a1>α6, and the expression of α3 and α5 had no difference (P>0. 05). In the subunit β family, the expression rank was β3>β2>β1. In the subunit γ family, the expression rank was γ1>γ2>γ3. (2) The expression of all subunits of GABAa receptor was obviously fluctuated excepting subunit α5 after treatment with SS. At 15 min post-SS, α1, α2 , β1 and γ1-3 were upregulated, and α3 was downregulated; At 30 min post-SS, α3, β1 and β3 were upregulated, and γ1 was downregulated; At 1 h post-SS, β2 was upregulated and γ3 was downregulated; At 3 h post-SS, β1 and β2 were upregulated, and α3 and γ2 were downregulated; At 6 h post-SS, αl, α3 ,β2, β3 and γ1 were upregulated, and α2, α4 and β1 were downregulated.
CONCLUSION
The mRNA of GABAaR was expressed in the rat SGN, and the expression of GABAaR subunits was lower in SGN than the cerebral cortex. SS could alter the GABAaR expression quantity in rat SGN; Most of the subunits expression were elevated obviously in the early post SS (15 min), followed by a slight fluctuation.
Animals
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Cells, Cultured
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Cochlea
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cytology
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In Situ Hybridization
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Neurons
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drug effects
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RNA, Messenger
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Rats
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Receptors, GABA-A
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metabolism
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Sodium Salicylate
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pharmacology
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Spiral Ganglion
;
drug effects
6.Mechanisms of muscovite on gastric mucosal protective effect.
Yun QIAN ; Jian-Min SI ; Liang-Jing WANG ; Shu-Jie CHEN ; You-Fa ZHU
China Journal of Chinese Materia Medica 2004;29(8):781-785
OBJECTIVETo explore the mechanisms of muscovite gastric mucosal protective effect.
METHODRat model of chronic gastritis were used. After gastric mucosal injury was induced, the rats were divided into 6 groups and were treated with different drugs. 2 weeks later, the tissue and blood samples were obtained and measured.
RESULTThe general conditions, the observations under macroscopy, microscope and electron microscope of the middle and high dose of muscovite groups resembled those of the normal group. Their PH levels were higher than those of the model group, and the rates of intestinal metaplasia were lower, but the PGE2 level of the middle dose of muscovite group was the highest.
CONCLUSIONMuscovite can be adsorbed on the surface of the gastric mucosa. It has gastric mucosal protective effect by improving excretion of mucus and synthesis of PGE2 in gastric mucosa, restraining gastric acid, reversing of intestinal metaplasia and decreasing inflammation cells.
Aluminum Compounds ; pharmacology ; Animals ; Dinoprostone ; blood ; Gastric Juice ; chemistry ; Gastric Mucosa ; pathology ; ultrastructure ; Gastritis ; blood ; chemically induced ; pathology ; Hydrogen-Ion Concentration ; Materia Medica ; pharmacology ; Microscopy, Electron, Scanning ; Potassium Compounds ; pharmacology ; Protective Agents ; pharmacology ; Rats ; Rats, Wistar ; Silicates ; pharmacology ; Sodium Salicylate
7.A study on toxic effects of sodium salicylate on rat cochlear spiral ganglion neurons: dopamine receptors mediate expressions of NMDA and GABAreceptors.
Ting-Jia WEI ; Hui-Ying CHEN ; Xi HUANG ; Jing-Jin WENG ; Jiang-Yuan QIN ; Ji-Ping SU
Acta Physiologica Sinica 2017;69(3):285-290
The aim of the present study was to observe whether dopamine receptor (DR) was involved in the effects of sodium salicylate (SS) on the expressions of N-methyl-D-aspartic acid (NMDA) and γ-aminobutyric acid (GABA) receptors in rat cochlear spiral ganglion neurons (SGNs). Forty-eight hours after primary culture of rat SGNs, immunofluorescence technique was applied to detect expressions of DR1 and DR2, the two subtypes of dopamine receptors. Western blot was performed to assess NMDA receptor NR1 subunit and GABAreceptor subunit α2 (GABRα2) protein expressions in the SGNs after the treatments of SS alone or in combination with DR antagonists. The results demonstrated that: (1) The DR1 and DR2 were expressed in the bodies and axons of the SGN; (2) After the treatment with SS, the surface protein expressions of GABRα2 and NR1 were decreased by 44.69% and 21.57%, respectively, while the total protein expressions showed no significant changes; (3) Neither SS + SCH23390 (DR1 antagonist) group nor SS + Eticlopride (DR2 antagonist) group showed significant differences in GABRα2 and NR1 surface protein expressions compared with the control group. These results suggest that SS regulates the surface GABAand NMDA receptors trafficking on SGN, and the mechanism may involve DR mediation.
Animals
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Benzazepines
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pharmacology
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Cells, Cultured
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Cochlea
;
cytology
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Neurons
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drug effects
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Rats
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Receptors, Dopamine
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metabolism
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Receptors, GABA-A
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metabolism
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Receptors, N-Methyl-D-Aspartate
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metabolism
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Sodium Salicylate
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toxicity
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Spiral Ganglion
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drug effects
8.The effects of sodium salicylate on the expression of GABAalpha, NR1 and hearing response properties of inferior colliculus neurons in mice.
Shi-Hua YIN ; Shu-Sheng GONG ; Kai-Sheng YAN ; Sui LI ; Pei CHEN ; Guang Li CHEN
Chinese Journal of Applied Physiology 2006;22(2):200-205
AIMTo study the effects of sodium salicylate on the expression of GABAalpha NR1 and hearing response properties of inferior colliculus neurons in mice.
METHODSThirty-six kunming mice were divided into three groups (A, B, C,). The expression of GABAalpha NR1 were measured by using RT-PCR. The intensity-rates functions, intensity-latency functions and frequency-turning curves were recorded by extracellular electrophysiological recording techniques.
RESULTS(1) The expression of GABAalpha mRNA of B group was decreased remarkably than the control group (A group, P < 0.05), there weren't noticeable differences between A group and C group (P > 0.05). The expression of NR1 mRNA of B group was increased remarkably than the control group (A group, P < 0.01), there were noticeable differences between A group and C group P < 0.05). (2) The intensity-rates functions, intensity-latency functions were monotonic while the frequency-turning curves were more broad when sodium salicylate was given. (3) The intensity-rates functions, intensity-latency functions were non-monotonic while the frequency-turning curves were sharpened after lidocaine was given.
CONCLUSIONS(1) The results suggested that administration of sodium salicylate decreased the expression of GABAalpha while increased the expression of NR1mRNA. (2) The intensity-rates functions, intensity-latency functions were monotonic, the frequency-turning curves were more broad when salicylate was given and the changes above could be reversed by given lidocaine.
Acoustic Stimulation ; Animals ; Inferior Colliculi ; drug effects ; metabolism ; physiology ; Mice ; Mice, Inbred Strains ; Neurons ; drug effects ; metabolism ; physiology ; Receptors, N-Methyl-D-Aspartate ; metabolism ; Sodium Salicylate ; pharmacology ; gamma-Aminobutyric Acid ; metabolism
9.Effects of sodium salicylate on the expressions of gamma-aminobutyricacid and glutamate and auditory response properties of the inferior colliculus neurons.
Shi-Hua YIN ; An-Zhou TANG ; Xiao-Ling XING ; Song-Hua TAN ; Li-Hong XIE
Acta Physiologica Sinica 2006;58(5):449-455
The effects of sodium salicylate (NaSA) on the expressions of gamma-aminobutyricacid (GABA) and glutamate (Glu), and auditory response properties of the inferior colliculus neurons in mice were studied. Thirty-six Kunming mice were divided into three groups: control group (saline injection); NaSA group (NaSA 450 mg/kg, i.p., each day for 15 d); NaSA + lidocaine group (NaSA 450 mg/kg + lidocaine 10 mg/kg, i.p., each day for 15 d). The expressions of GABA and Glu were examined with immunohistochemical method. The intensity-rate function, intensity-latency function and frequency-tuning curve were determined with extracellular electrophysiological recording. Results are as follows: (1) The expression of GABA in the NaSA and NaSA + lidocaine groups decreased remarkably compared with that in the control group; there was no noticeable difference between the NaSA and NaSA + lidocaine groups. The expression of Glu in the NaSA group increased significantly compared with that in the control and NaSA + lidocaine groups. No difference in the expression of Glu was found between the control and NaSA + lidocaine groups. (2) In NaSA group, the intensity-rate function displayed a non-monotonic pattern, rising at low intensity and descending at high intensity; the tip of frequency-tuning curves became broad after administration of NaSA. (3) The changes in intensity-rate function and intensity-latency function were not evident and the tips of the frequency-tuning curves sharpened in the NaSA + lidocaine group. These results suggest that administration of NaSA increases the expression of Glu-positive neurons and reduces that of GABA-positive neurons in the inferior colliculus. NaSA changes the auditory response properties of the inferior colliculus and lidocaine can reverse these changes.
Acoustic Stimulation
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Animals
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Female
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Glutamates
;
analysis
;
Glutamic Acid
;
analysis
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Immunohistochemistry
;
Inferior Colliculi
;
chemistry
;
drug effects
;
physiology
;
Male
;
Mice
;
Reaction Time
;
drug effects
;
Sodium Salicylate
;
pharmacology
;
gamma-Aminobutyric Acid
;
analysis
10.Effect of sodium para-aminosalicylic on concentrations of amino acid neurotransmitters in basal ganglia of manganese-exposed rats.
Chao-yan OU ; Ming-li HUANG ; Yue-ming JIANG ; Hai-lan LUO ; Xiang-fa DENG ; Chan WANG ; Fang WANG ; Xiao-wei HUANG
Chinese Journal of Preventive Medicine 2011;45(5):422-425
OBJECTIVETo probe the effect of sodium para-aminosalicylate (PAS-Na) on concentration of amino acid neurotransmitters including glutamate (Glu), glutamine (Gln), glycine (Gly) and gamma-aminobutyric acid (GABA) in basal ganglia of subacute manganese (Mn)-exposed rats.
METHODSForty Sprague-Dawley male rats were randomly divided into the control, Mn-exposed, low dose PAS-Na (L-PAS) and high dose PAS-Na (H-PAS) groups. Rats in experiment groups received daily intraperitoneally injections of manganese chloride (MnCl₂ · 4H₂O, 15 mg/kg), while rats in control group received daily intraperitoneally injections of normal saline (NS), all at 5 days/week for 4 weeks. Then the rats in PAS groups followed by a daily subcutaneously dose of PAS-Na (100 and 200 mg/kg as the L-PAS and H-PAS groups, respectively) for another 3 and 6 weeks; while the rats in Mn-exposed and control group received NS. The concentrations of Glu, Gln, Gly and GABA in basal ganglia of rat was detected by the high performance liquid chromatography fluorescence detection technique.
RESULTSAfter treating with PAS-Na for 3 weeks, the concentration of Gly in the Mn-exposed rats decreased to (0.165 ± 0.022) µmol/L (control = (0.271 ± 0.074) µmol/L, Mn vs control, t = 4.65, P < 0.05). After the further 6-week therapy with PAS-Na, the concentrations of Glu, Gln, Gly in the Mn-exposed rats were lower than those of the control rats ((0.942 ± 0.121), (0.377 ± 0.070), (0.142 ± 0.048), (1.590 ± 0.302), (0.563 ± 0.040), (0.247 ± 0.084) µmol/L; t = 7.72, 5.85, 4.30, P < 0.05); and also lower than in L-PAS and H-PAS groups, whose concentrations were separately (1.268 ± 0.124), (1.465 ± 0.196), (0.497 ± 0.050), (0.514 ± 0.103), (0.219 ± 0.034) µmol/L (L-PAS Glu and Gln vs Mn, t = 3.87, 3.77, P < 0.05; H-PAS Glu, Gln and Gly vs Mn, t = 6.78, 4.70, 3.42, P < 0.05).
CONCLUSIONThe toxic effect of manganese on Glu, Gln and Gly in basal ganglia of Mn-exposed rats is obvious, especially appears earlier on Gly. The toxic effect still continues to develop when relieved from the exposure. PAS-Na may play an antagonism role in toxic effect of manganese on concentration of Glu, Gln and Gly in basal ganglia of Mn-exposed rats.
Amino Acids ; metabolism ; Animals ; Basal Ganglia ; drug effects ; metabolism ; Glutamic Acid ; metabolism ; Male ; Manganese ; toxicity ; Neurotransmitter Agents ; metabolism ; Rats ; Rats, Sprague-Dawley ; Sodium Salicylate ; pharmacology ; gamma-Aminobutyric Acid ; metabolism