耳穴电刺激配合声音掩蔽对耳鸣大鼠听觉皮层的CREB蛋白、BDNF及其受体TrkB表达的影响
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摘要
目的:探讨耳穴电刺激配合声音掩蔽对耳鸣大鼠听觉皮层的环磷酸腺苷反应元件结合蛋白(CREB)、脑源性神经营养因子(BDNF)及其受体酪氨酸蛋白激酶受体B(TrkB)表达的影响。方法:将27只成年雄性SD大鼠随机分为对照组、模型组和治疗组。模型组及治疗组大鼠采取腹腔注射水杨酸钠诱导制作耳鸣模型;对照组注射0.9%NaCl溶液。造模成功后,治疗组采用耳穴"神门""胰胆"电刺激配合声音掩蔽法治疗,每天1次,治疗15d。采用声音惊吓反应监测系统监测各组大鼠的停顿声音预刺激抑制惊吓反应(GPIAS)及预刺激抑制反应(PPI),以WesternBlot法检测各组大鼠听觉皮层的BDNF及其受体TrkB,CREB和磷酸化CREB(p-CREB)蛋白表达。结果:①与对照组比较,模型组大鼠在12、16、20、28 kHz高频背景声音的GPIAS比值均下降(均P<0.05);与模型组比较,治疗组在12、16、20、28 kHz高频背景声音的GPIAS比值均升高(均P<0.05)。②与对照组比较,模型组BDNF蛋白表达、p-CREB蛋白表达均升高(均P<0.01),TrkB表达升高(P<0.05);治疗组与模型组BDNF和TrkB表达、CREB及p-CREB蛋白表达比较差异均无统计学意义(均P>0.05)。结论:耳穴电刺激配合声音掩蔽法能改善耳鸣大鼠高频背景声音的GPIAS比值变化,此作用可能与抑制耳鸣大鼠听觉皮层的BDNF/TrkB/CREB信号通路相关蛋白表达下调,进而逆转不良可塑性改变有关。
Objective To explore the effect of electrical stimulation at auricular points(EAS) combined with sound masking on the expression of cAMP-response element binding protein(CREB), brain-derived neurotrophic factor(BDNF) and tyrosine receptor kinase B(TrkB) in the auditory cortex of tinnitus rats. Methods A total of 27 adult male SD rats were randomly divided into a control group, a model group and an EAS group. The rats in the model group and the EAS group were intervened with intraperitoneal injection of sodium salicylate to induce tinnitus model, while the rats in the control group were intervened with injection of 0.9% NaCl solution. After the model was successfully established, the rats in the EAS group were treated with electrical stimulation at "Shenmen"(TF_4) and "Yidan"(CO_(11)),combined with sound masking; the treatment was given once a day for 15 days. The gap prepulse inhibition of acoustic startle(GPIAS) and prepulse inhibition(PPI) testing were performed using the acoustic startle reflex starter package for rats. The expression of BDNF, TrkB, CREB and p-CREB in the auditory cortex of each group were measured with Western Blot analysis. Results ① Compared with the control group, the GPIAS values in 12 kHz, 16 kHz, 20 kHz and28 kHz were significantly decreased in the model group(all P<0.05); compared with the model group,GPIAS values in 12 kHz, 16 kHz, 20 kHz and 28 kHz were significantly increased in the EAS group(all P<0.05).② Compared with the control group, the expression of BDNF and p-CREB in the model group was significantly increased(P<0.01), and the expression of TrkB in the model group was significantly increased(P<0.05); the differences of expression of BDNF, TrkB, CREB and p-CREB between the model group and the EAS group had no statistics significance(all P>0.05). Conclusion EAS could improve the GPIAS values of high-frequency background sound in tinnitus rats, which may be related with the upregulation of the BDNF/TrkB/CREB signaling pathway in the auditory cortex, leading to the reversion of the maladaptive plasticity.
Objective To explore the effect of electrical stimulation at auricular points(EAS) combined with sound masking on the expression of cAMP-response element binding protein(CREB), brain-derived neurotrophic factor(BDNF) and tyrosine receptor kinase B(TrkB) in the auditory cortex of tinnitus rats. Methods A total of 27 adult male SD rats were randomly divided into a control group, a model group and an EAS group. The rats in the model group and the EAS group were intervened with intraperitoneal injection of sodium salicylate to induce tinnitus model, while the rats in the control group were intervened with injection of 0.9% NaCl solution. After the model was successfully established, the rats in the EAS group were treated with electrical stimulation at "Shenmen"(TF_4) and "Yidan"(CO_(11)),combined with sound masking; the treatment was given once a day for 15 days. The gap prepulse inhibition of acoustic startle(GPIAS) and prepulse inhibition(PPI) testing were performed using the acoustic startle reflex starter package for rats. The expression of BDNF, TrkB, CREB and p-CREB in the auditory cortex of each group were measured with Western Blot analysis. Results ① Compared with the control group, the GPIAS values in 12 kHz, 16 kHz, 20 kHz and28 kHz were significantly decreased in the model group(all P<0.05); compared with the model group,GPIAS values in 12 kHz, 16 kHz, 20 kHz and 28 kHz were significantly increased in the EAS group(all P<0.05).② Compared with the control group, the expression of BDNF and p-CREB in the model group was significantly increased(P<0.01), and the expression of TrkB in the model group was significantly increased(P<0.05); the differences of expression of BDNF, TrkB, CREB and p-CREB between the model group and the EAS group had no statistics significance(all P>0.05). Conclusion EAS could improve the GPIAS values of high-frequency background sound in tinnitus rats, which may be related with the upregulation of the BDNF/TrkB/CREB signaling pathway in the auditory cortex, leading to the reversion of the maladaptive plasticity.
引文
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[5]杨松柏,梅志刚,谭凌菁,等.耳迷走神经分布区穴位电刺激配合声音掩蔽法对耳鸣大鼠听性脑干反应及下丘神经递质的影响[J].中国针灸,2016,36(5):517-522.
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[12]Zhao J,Wang B,Wang X,et al.Up-regulation of Ca2+/Ca MKII/CREB signaling in salicylate-induced tinnitus in rats[J].Mol Cell Biochem,2018,448(1-2):71-76.doi:10.1007/s11010-018-3314-z.
[13]Yi B,Wu C,Shi R,et al.Long-term administration of salicylate-induced changes in BDNF expression and CREBphosphorylation in the auditory cortex of rats[J].Otol Neurotol,2018,39(3):e173-e180.
[14]Coskunoglu A,Orenay-Boyacioglu S,Deveci A,et al.Evidence of associations between brain-derived neurotrophic factor(BDNF)serum levels and gene polymorphisms with tinnitus[J].Noise Health,2017,19(88):140-148.
[15]Xiong H,Yang H,Liang M,et al.Plasma brain-derived neurotrophic factor levels are increased in patients with tinnitus and correlated with therapeutic effects[J].Neurosci Lett,2016,622:15-18.
[16]Goto F,Saruta J,Kanzaki S,et al.Various levels of plasma brain-derived neurotrophic factor in patients with tinnitus[J].Neurosci Lett,2012,510(2):73-77.
[17]Rasmussen P,Brassard P,Adser H,et al.Evidence for a release of brain-derived neurotrophic factor from the brain during exercise[J].Exp Physiol,2009,94(10):1062-1069.
[18]吴莎,华清泉,杨琨,等.脑源性神经营养因子研究进展[J].中华临床医师杂志(电子版),2013,7(9):3988-3990.
[19]Lu B.BDNF and acigtivity-dependent synaptic modulation[J].Learn Mem,2003,10(2):86-89.
[20]Saura CA,Valero J.The role of CREB signaling in Alzheimer’s disease and other cognitive disorders[J].Rev Neurosci,2011,22(2):153-169.
[21]Obrietan K,Gao XB,Van Den Pol AN.Excitatory actions of GABA increase BDNF expression via a MAPK-CREB-dependent mechanism-a positive feedback circuit in developing neurons[J].JNeurophysiol,2002,88(2):1005-1015.
[22]Rivera C,Li H,Thomas-Crusells J,et al.BDNF-induced TrkBactivation down-regulates the K+-Cl-cotransporter KCC2 and impairs neuronal Cl-extrusion[J].J Cell Biol,2002,159(5):747-752.
[23]王玉杏,徐鸥,刘砚星,等.听觉剥夺大鼠听皮层BDNF和Trk B的表达变化[J].听力学及言语疾病杂志,2013,21(4):379-382.
[24]Tziridis K,Ahlf S,Schulze H.A low cost setup for behavioral audiometry in rodents[J].J Vis Exp,2012(68):pii:4433.doi:10.3791/4433.
[25]Ouyang J,Pace E,Lepczyk L,et al.Blast-induced tinnitus and elevated central auditory and limbic activity in rats:a manganese-enhanced MRI and behavioral study[J].Sci Rep,2017,7(1):4852.doi:10.1038/s41598-017-04941-w.
[26]Guilloux JP,Douillard-Guilloux G,Kota R,et al.Molecular evidence for BDNF-and GABA-related dysfunctions in the amygdala of female subjects with major depression[J].Mol Psychiatry,2012,17(11):1130-1142.
[2]尹志华,马灵草,刘宏建,等.耳鸣人群流行病学特征分析[J].医药论坛杂志,2015,36(1):55-59.
[3]Irvine DRF.Plasticity in the auditory system[J].Hear Res,2018,362:61-73.
[4]Mei ZG,Yang SB,Cai SJ,et al.Treatment of tinnitus with electrical stimulation on acupoint in the distribution area of ear vagus nerve combining with sound masking:randomized controlled trial[J].World J Acup-Moxi,2014,24(2):30-35.
[5]杨松柏,梅志刚,谭凌菁,等.耳迷走神经分布区穴位电刺激配合声音掩蔽法对耳鸣大鼠听性脑干反应及下丘神经递质的影响[J].中国针灸,2016,36(5):517-522.
[6]Jastreboff PJ,Brennan JF,Coleman JK,et al.Phantom auditory sensation in rats:an animal model for tinnitus[J].Behav Neurosci,1988,102(6):811-822.
[7]Turner JG,Brozoski TJ,Bauer CA,et al.Gap detection deficits in rats with tinnitus:a potential novel screening tool[J].Behav Neurosci,2006,120(1):188-195.
[8]Shore SE,Roberts LE,Langguth B.Maladaptive plasticity in tinnitus-triggers,mechanisms and treatment[J].Nat Rev Neurol,2016,12(3):150-160.
[9]Hu SS,Mei L,Chen JY,et al.Effects of salicylate on the inflammatory genes expression and synaptic ultrastructure in the cochlear nucleus of rats[J].Inflammation,2014,37(2):365-373.
[10]Hu SS,Mei L,Chen JY,et al.Expression of immediate early genes in the dorsal cochlear nucleus in salicylate induced tinnitus[J].Eur Arch Otorhinolaryngol,2016,273(2):325-332.
[11]王洪田,孙建和,于宁水,等.杨酸钠引起耳鸣大鼠听皮层突触形态改变的透射电镜观察[J].中华耳科学杂志,2009,7(3):208-211.
[12]Zhao J,Wang B,Wang X,et al.Up-regulation of Ca2+/Ca MKII/CREB signaling in salicylate-induced tinnitus in rats[J].Mol Cell Biochem,2018,448(1-2):71-76.doi:10.1007/s11010-018-3314-z.
[13]Yi B,Wu C,Shi R,et al.Long-term administration of salicylate-induced changes in BDNF expression and CREBphosphorylation in the auditory cortex of rats[J].Otol Neurotol,2018,39(3):e173-e180.
[14]Coskunoglu A,Orenay-Boyacioglu S,Deveci A,et al.Evidence of associations between brain-derived neurotrophic factor(BDNF)serum levels and gene polymorphisms with tinnitus[J].Noise Health,2017,19(88):140-148.
[15]Xiong H,Yang H,Liang M,et al.Plasma brain-derived neurotrophic factor levels are increased in patients with tinnitus and correlated with therapeutic effects[J].Neurosci Lett,2016,622:15-18.
[16]Goto F,Saruta J,Kanzaki S,et al.Various levels of plasma brain-derived neurotrophic factor in patients with tinnitus[J].Neurosci Lett,2012,510(2):73-77.
[17]Rasmussen P,Brassard P,Adser H,et al.Evidence for a release of brain-derived neurotrophic factor from the brain during exercise[J].Exp Physiol,2009,94(10):1062-1069.
[18]吴莎,华清泉,杨琨,等.脑源性神经营养因子研究进展[J].中华临床医师杂志(电子版),2013,7(9):3988-3990.
[19]Lu B.BDNF and acigtivity-dependent synaptic modulation[J].Learn Mem,2003,10(2):86-89.
[20]Saura CA,Valero J.The role of CREB signaling in Alzheimer’s disease and other cognitive disorders[J].Rev Neurosci,2011,22(2):153-169.
[21]Obrietan K,Gao XB,Van Den Pol AN.Excitatory actions of GABA increase BDNF expression via a MAPK-CREB-dependent mechanism-a positive feedback circuit in developing neurons[J].JNeurophysiol,2002,88(2):1005-1015.
[22]Rivera C,Li H,Thomas-Crusells J,et al.BDNF-induced TrkBactivation down-regulates the K+-Cl-cotransporter KCC2 and impairs neuronal Cl-extrusion[J].J Cell Biol,2002,159(5):747-752.
[23]王玉杏,徐鸥,刘砚星,等.听觉剥夺大鼠听皮层BDNF和Trk B的表达变化[J].听力学及言语疾病杂志,2013,21(4):379-382.
[24]Tziridis K,Ahlf S,Schulze H.A low cost setup for behavioral audiometry in rodents[J].J Vis Exp,2012(68):pii:4433.doi:10.3791/4433.
[25]Ouyang J,Pace E,Lepczyk L,et al.Blast-induced tinnitus and elevated central auditory and limbic activity in rats:a manganese-enhanced MRI and behavioral study[J].Sci Rep,2017,7(1):4852.doi:10.1038/s41598-017-04941-w.
[26]Guilloux JP,Douillard-Guilloux G,Kota R,et al.Molecular evidence for BDNF-and GABA-related dysfunctions in the amygdala of female subjects with major depression[J].Mol Psychiatry,2012,17(11):1130-1142.