电针对面神经损伤后面神经元中神经型钙黏素、上皮型钙黏素和胎盘型钙黏素表达的影响
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摘要
背景:作者所在课题组前期研究发现周围性面神经损伤能引起神经元中黏附分子的表达变化,而电针穴位治疗周围性面神经损伤的机制尚不清楚。目的:探讨电针穴位在面神经外周部受损后对中枢面神经元的形态学变化及面神经元中神经型钙黏素、上皮型钙黏素及胎盘型钙黏素表达的影响。方法:将新西兰大白兔随机分为正常对照组(n=4)、面神经损伤组(n=24)和电针组(n=24)。面神经损伤组与电针组均建立右侧面神经上颊支压榨损伤模型。造模成功后每天对电针组实验兔颊车、地仓、翳风、颧缪、四白、阳白电刺激,选择频率为18-20 Hz、电压为1.5 V的疏密波,另外埋针合谷穴不予以电刺激,针灸每次30 min,1次/d。结果与结论:(1)原位杂交结果显示各组兔面神经元中均未见上皮型钙黏素和胎盘型钙黏素mRNA表达,且正常对照组兔面神经元中未见神经型钙黏素mRNA表达,而面神经损伤组和电针组兔面神经元中出现神经型钙黏素mRNA的阳性表达,电针组兔面神经元中神经型钙黏素mRNA阳性细胞数均多于面神经损伤组(P<0.05);(2)免疫组化染色可见正常对照组兔面神经元中神经型钙黏素、上皮型钙黏素和胎盘型钙黏素蛋白表达;损伤后1 d时三者表达下降(电针组神经型钙黏素蛋白除外);损伤后1,4,14 d时电针组兔面神经元中神经型钙黏素和胎盘型钙黏素蛋白表达高于面神经损伤组(P<0.05);损伤后各时间点,电针组上皮型钙黏素蛋白表达均高于面神经损伤组(1 d时除外)(P<0.05);(3)苏木精-伊红染色可见面神经损伤组逐渐出现面神经元变性及类似神经元死亡表现,21 d时神经元开始恢复。电针组变性及类似死亡表现的神经元较面神经损伤组少,且14 d时神经元开始恢复,28 d时2组神经元形态均基本恢复;(4)提示面神经损伤引起了钙黏素的表达变化,电针治疗减轻了面神经元的变性并促进神经元恢复,这可能与电针促进面神经元中经典钙黏素的高表达密切相关。
BACKGROUND: Preliminary studies have found that peripheral facial nerve injury can cause changes in the expression of adhesion molecules in neurons, but the mechanism of electroacupuncture in the treatment of facial nerve injury is unclear. OBJECTIVE: To observe the effect of electroacupuncture on the morphological changes of the facial neurons and the expression of N-cadherin, E-cadherin and P-cadherin in the neurons after peripheral facial nerve injury. METHODS: New Zealand white rabbits were randomized into control group(n=4), model group(n=24) and electroacupuncture group(n=24). Rabbits in the latter two groups were used to establish the crush injury model of superior buccal branch of the right facial nerve, and the electroacupuncture group rabbits were given the electroacupuncture at Jiache(ST6), Dicang(ST4), Yifeng(SJ17), Quanliao(SI18), Sibai(ST2) and Yangbai(GB14) with dilatational wave(18-20 Hz, 1.5 V), and needle-embedding at Hegu(LI4) without electroacupuncture, 30 minutes, once daily. RESULTS AND CONCLUSION: In situ hybridization results suggested that E-cadherin mRNA and P-cadherin mRNA were negative in all groups, and N-cadherin mRNA was not expressed in the control group. The expression of N-cadherin mRNA was positive in the electroacupuncture and model groups after facial nerve injury. The number of N-cadherin mRNA positive cells in the electroacupuncture group was significantly higher than that in the model group(P < 0.05). Immunohistochemical staining results found that the expressions of N-cadherin, E-cadherin and P-cadherin were positive in the control group; the expression levels of the three cadherins decreased at 1 day after injury(except N-cadherin in the electroacupuncture group); the expression levels of N-cadherin and P-cadherin in the electroacupuncture group were higher than those in the model group at 1, 4 and 14 days after injury(P < 0.05); the expression levels of E-cadherin in the electroacupuncture group was significantly higher than that in the model group(except the first day)(P < 0.05). Hematoxylin-eosin staining showed that in the model group, there was facial nerve neuron degeneration followed by similar neuronal death, and the neuron morphology began to recover at 21 days after injury. Neuron morphological changes in the electroacupuncture group were less than those in the model group, and neurons began to recover at 14 days after injury. The neurons in the two groups were restored at 28 days after injury. Our findings reveal that facial nerve injury can induce the expression of adhesion molecules, and electroacupuncture can reduce the degeneration of facial neurons and promote the recovery of neurons, which may be achieved by increasing the expression of adhesion molecules in the facial neurons.
BACKGROUND: Preliminary studies have found that peripheral facial nerve injury can cause changes in the expression of adhesion molecules in neurons, but the mechanism of electroacupuncture in the treatment of facial nerve injury is unclear. OBJECTIVE: To observe the effect of electroacupuncture on the morphological changes of the facial neurons and the expression of N-cadherin, E-cadherin and P-cadherin in the neurons after peripheral facial nerve injury. METHODS: New Zealand white rabbits were randomized into control group(n=4), model group(n=24) and electroacupuncture group(n=24). Rabbits in the latter two groups were used to establish the crush injury model of superior buccal branch of the right facial nerve, and the electroacupuncture group rabbits were given the electroacupuncture at Jiache(ST6), Dicang(ST4), Yifeng(SJ17), Quanliao(SI18), Sibai(ST2) and Yangbai(GB14) with dilatational wave(18-20 Hz, 1.5 V), and needle-embedding at Hegu(LI4) without electroacupuncture, 30 minutes, once daily. RESULTS AND CONCLUSION: In situ hybridization results suggested that E-cadherin mRNA and P-cadherin mRNA were negative in all groups, and N-cadherin mRNA was not expressed in the control group. The expression of N-cadherin mRNA was positive in the electroacupuncture and model groups after facial nerve injury. The number of N-cadherin mRNA positive cells in the electroacupuncture group was significantly higher than that in the model group(P < 0.05). Immunohistochemical staining results found that the expressions of N-cadherin, E-cadherin and P-cadherin were positive in the control group; the expression levels of the three cadherins decreased at 1 day after injury(except N-cadherin in the electroacupuncture group); the expression levels of N-cadherin and P-cadherin in the electroacupuncture group were higher than those in the model group at 1, 4 and 14 days after injury(P < 0.05); the expression levels of E-cadherin in the electroacupuncture group was significantly higher than that in the model group(except the first day)(P < 0.05). Hematoxylin-eosin staining showed that in the model group, there was facial nerve neuron degeneration followed by similar neuronal death, and the neuron morphology began to recover at 21 days after injury. Neuron morphological changes in the electroacupuncture group were less than those in the model group, and neurons began to recover at 14 days after injury. The neurons in the two groups were restored at 28 days after injury. Our findings reveal that facial nerve injury can induce the expression of adhesion molecules, and electroacupuncture can reduce the degeneration of facial neurons and promote the recovery of neurons, which may be achieved by increasing the expression of adhesion molecules in the facial neurons.
引文
[1]田勇泉.耳鼻咽喉-头颈外科学[M].8版.北京:人民卫生出版社,2013.
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[3]卫彦,寇吉友.电针对兔实验性周围性面瘫干预作用量效关系的影响[J].上海针灸杂志,2014,33(6):589-591.
[4]李雷激,骆文龙,曾卫东,等.穴位电针对面神经损伤后面神经核中黏附分子CD56表达的影响[J].重庆医学,2006,35(24):2239-2241.
[5]牟鸿.单核细胞趋化蛋白-1?细胞间粘附分子-1在病毒性面瘫小鼠脑干面神经核团的动态表达及意义[D].济南:山东大学,2015.
[6]牙祖蒙,王建华,李忠禹,等.面神经损伤后穴位电针刺激对面神经核中几种神经营养因子m RNA表达的影响[J].中国中西医结合耳鼻咽喉科杂志,2001,9(5):209-211.
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[10]孙力超.结肠癌肝转移诊断?治疗相关分子靶标的研究[D].北京:中国医学科学院,2009.
[11]Gwak GY,Yoon JH,Yu SJ,et al.Anti-apoptotic N-cadherin signaling and its prognostic implication in human hepatocellular carcinomas.Oncol Rep.2006;15(5):1117-1123.
[12]Lin S,Stonyanova T,Kono E,et al.Abstract LB-279:N-cadherin promotes castration-resistant prostate cancer progression by enhancing stem cell properties of prostate cancer cells.Cancer Res.2016;76(14 Supplement):LB-279.
[13]Horne HN,Oh H,Sherman ME,et al.Abstract 3451:Breast cancer risk factor associations by loss of E-cadherin tumor tissue expression:Apooled analysis of 5,896 cases in 12 studies from the Breast Cancer Association Consortium(BCAC).Cancer Res.2016;76(14 Supplement):3451.
[14]Li B,Shi H2,Wang F1,et al.Expression of E-,P-and N-Cadherin and Its Clinical Significance in Cervical Squamous Cell Carcinoma and Precancerous Lesions.PLo S One.2016;11(5):e0155910.
[15]Rebman JK,Kirchoff KE,Walsh GS.Cadherin-2 Is Required Cell Autonomously for Collective Migration of Facial Branchiomotor Neurons.PLo S One.2016;11(10):e0164433.
[16]Xu C,Funahashi Y,Watanabe T,et al.Radial Glial Cell-Neuron Interaction Directs Axon Formation at the Opposite Side of the Neuron from the Contact Site.J Neurosci.2015;35(43):14517-14532.
[17]Liu Q,Londraville RL,Azodi E,et al.Up-regulation of cadherin-2 and cadherin-4 in regenerating visual structures of adult zebrafish.Exp Neurol.2002;177(2):396-406.
[18]Hatoko M,Tada H,Tanaka A,et al.The differential expression of N-cadherin in vascularized and nonvascularized nerve grafts:a study in a rat sciatic nerve model.Ann Plast Surg.2001;47(3):322-327.
[19]Tada H,Hatoko M,Tanaka A,et al.The difference in E-cadherin expression between nonvascularized and vascularized nerve grafts:study in the rat sciatic nerve model.J Surg Res.2001;100(1):57-62.
[20]牙祖蒙,王建华,李忠禹,等.面神经损伤后穴位电针刺激对神经组织中神经营养因子-3及其受体表达的影响[J].中国中医基础医学杂志,2000,6(1):62-65.
[21]Paxinos G,Watson C.The Rat Brain Stereotaxic Coordinates.Sydney:Reed Elsevier Group PLC.2007:166-167.
[22]李雷激,徐超然,覃纲,等.面神经损伤后面神经核中神经型钙黏附分子及胎盘型钙黏附分子的表达[J].中国组织工程研究,2015,19(37):5978-5982.
[23]于亚楠,刘彦礼,徐振平,等.视网膜钙黏蛋白在中枢神经系统发育中的作用研究现状[J].新乡医学院学报,2015,32(9):804-806.
[24]Paulson AF,Prasad MS,Thuringer AH,et al.Regulation of cadherin expression in nervous system development.Cell Adh Migr.2014;8(1):19-28.
[25]Astick M,Tubby K,Mubarak WM,et al.Central topography of cranial motor nuclei controlled by differential cadherin expression.Curr Biol.2014;24(21):2541-2547.
[26]Redies C,Takeichi M.Expression of N-cadherin m RNA during development of the mouse brain.Dev Dyn.1993;197(1):26-39.
[27]Scheib J,H?ke A.Advances in peripheral nerve regeneration.Nat Rev Neurol.2013;9(12):668-676.
[28]Faroni A,Mobasseri SA,Kingham PJ,et al.Peripheral nerve regeneration:experimental strategies and future perspectives.Adv Drug Deliv Rev.2015;82-83:160-167.
[29]Sun Z,Wei W,Liu H,et al.Acute Response of Neurons:An Early Event of Neuronal Cell Death After Facial Nerve Injury.World Neurosurg.2018;109:e252-e257.
[30]Zelano J,Wallquist W,Hailer NP,et al.Expression of nectin-1,nectin-3,N-cadherin,and NCAM in spinal motoneurons after sciatic nerve transection.Exp Neurol.2006;201(2):461-469.
[31]张晓晖.电针治疗对坐骨神经损伤大鼠NGF?CNTF?BDNF?b FGF表达的影响[D].北京:北京中医药大学,2011.
[2]李坚将,刘辉.针灸治疗面瘫及对口唇?甲襞微循环的影响[J].上海针灸杂志,2001,20(5):16-18.
[3]卫彦,寇吉友.电针对兔实验性周围性面瘫干预作用量效关系的影响[J].上海针灸杂志,2014,33(6):589-591.
[4]李雷激,骆文龙,曾卫东,等.穴位电针对面神经损伤后面神经核中黏附分子CD56表达的影响[J].重庆医学,2006,35(24):2239-2241.
[5]牟鸿.单核细胞趋化蛋白-1?细胞间粘附分子-1在病毒性面瘫小鼠脑干面神经核团的动态表达及意义[D].济南:山东大学,2015.
[6]牙祖蒙,王建华,李忠禹,等.面神经损伤后穴位电针刺激对面神经核中几种神经营养因子m RNA表达的影响[J].中国中西医结合耳鼻咽喉科杂志,2001,9(5):209-211.
[7]张微.电针对面神经损伤修复的雪旺细胞形态学及CNTF表达影响的研究[D].成都:成都中医药大学,2012.
[8]李雷激,骆文龙,伍宗惠,等.面神经损伤后面神经核中黏附分子CD44表达与穴位电针刺激的影响[J].中国组织工程研究与临床康复,2007,11(27):5369-5373.
[9]李斐,江普查,袁先厚,等.N-cadherin?E-cadherin及β-catenin在胶质瘤中的表达及意义[J].中国临床神经外科杂志,2007,12(9):536-539.
[10]孙力超.结肠癌肝转移诊断?治疗相关分子靶标的研究[D].北京:中国医学科学院,2009.
[11]Gwak GY,Yoon JH,Yu SJ,et al.Anti-apoptotic N-cadherin signaling and its prognostic implication in human hepatocellular carcinomas.Oncol Rep.2006;15(5):1117-1123.
[12]Lin S,Stonyanova T,Kono E,et al.Abstract LB-279:N-cadherin promotes castration-resistant prostate cancer progression by enhancing stem cell properties of prostate cancer cells.Cancer Res.2016;76(14 Supplement):LB-279.
[13]Horne HN,Oh H,Sherman ME,et al.Abstract 3451:Breast cancer risk factor associations by loss of E-cadherin tumor tissue expression:Apooled analysis of 5,896 cases in 12 studies from the Breast Cancer Association Consortium(BCAC).Cancer Res.2016;76(14 Supplement):3451.
[14]Li B,Shi H2,Wang F1,et al.Expression of E-,P-and N-Cadherin and Its Clinical Significance in Cervical Squamous Cell Carcinoma and Precancerous Lesions.PLo S One.2016;11(5):e0155910.
[15]Rebman JK,Kirchoff KE,Walsh GS.Cadherin-2 Is Required Cell Autonomously for Collective Migration of Facial Branchiomotor Neurons.PLo S One.2016;11(10):e0164433.
[16]Xu C,Funahashi Y,Watanabe T,et al.Radial Glial Cell-Neuron Interaction Directs Axon Formation at the Opposite Side of the Neuron from the Contact Site.J Neurosci.2015;35(43):14517-14532.
[17]Liu Q,Londraville RL,Azodi E,et al.Up-regulation of cadherin-2 and cadherin-4 in regenerating visual structures of adult zebrafish.Exp Neurol.2002;177(2):396-406.
[18]Hatoko M,Tada H,Tanaka A,et al.The differential expression of N-cadherin in vascularized and nonvascularized nerve grafts:a study in a rat sciatic nerve model.Ann Plast Surg.2001;47(3):322-327.
[19]Tada H,Hatoko M,Tanaka A,et al.The difference in E-cadherin expression between nonvascularized and vascularized nerve grafts:study in the rat sciatic nerve model.J Surg Res.2001;100(1):57-62.
[20]牙祖蒙,王建华,李忠禹,等.面神经损伤后穴位电针刺激对神经组织中神经营养因子-3及其受体表达的影响[J].中国中医基础医学杂志,2000,6(1):62-65.
[21]Paxinos G,Watson C.The Rat Brain Stereotaxic Coordinates.Sydney:Reed Elsevier Group PLC.2007:166-167.
[22]李雷激,徐超然,覃纲,等.面神经损伤后面神经核中神经型钙黏附分子及胎盘型钙黏附分子的表达[J].中国组织工程研究,2015,19(37):5978-5982.
[23]于亚楠,刘彦礼,徐振平,等.视网膜钙黏蛋白在中枢神经系统发育中的作用研究现状[J].新乡医学院学报,2015,32(9):804-806.
[24]Paulson AF,Prasad MS,Thuringer AH,et al.Regulation of cadherin expression in nervous system development.Cell Adh Migr.2014;8(1):19-28.
[25]Astick M,Tubby K,Mubarak WM,et al.Central topography of cranial motor nuclei controlled by differential cadherin expression.Curr Biol.2014;24(21):2541-2547.
[26]Redies C,Takeichi M.Expression of N-cadherin m RNA during development of the mouse brain.Dev Dyn.1993;197(1):26-39.
[27]Scheib J,H?ke A.Advances in peripheral nerve regeneration.Nat Rev Neurol.2013;9(12):668-676.
[28]Faroni A,Mobasseri SA,Kingham PJ,et al.Peripheral nerve regeneration:experimental strategies and future perspectives.Adv Drug Deliv Rev.2015;82-83:160-167.
[29]Sun Z,Wei W,Liu H,et al.Acute Response of Neurons:An Early Event of Neuronal Cell Death After Facial Nerve Injury.World Neurosurg.2018;109:e252-e257.
[30]Zelano J,Wallquist W,Hailer NP,et al.Expression of nectin-1,nectin-3,N-cadherin,and NCAM in spinal motoneurons after sciatic nerve transection.Exp Neurol.2006;201(2):461-469.
[31]张晓晖.电针治疗对坐骨神经损伤大鼠NGF?CNTF?BDNF?b FGF表达的影响[D].北京:北京中医药大学,2011.