激光扫描共聚焦显微镜技术在针灸形态学研究中的应用
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摘要
激光扫描共聚焦显微镜(LSCM)应用于针灸形态学方面的研究已经有20多年,并成为从组织学角度探讨腧穴结构和针灸效应的重要工具之一。借助于LSCM的技术优势,针灸形态学研究水平得到进一步提升,使我们对腧穴和经络相关的组织结构以及针灸引起的组织化学变化有了更加深入的了解。为了让LSCM在针灸形态学研究方面更有效地发挥作用,本文将对以往有关方面的研究成果进行全面的梳理,同时结合自己的实际经验在技术层面上给出具体建议。期待LSCM技术能够被整合到更多的针灸实验研究中,为揭示针灸的作用机制提供更有力的形态学依据。
Laser scanning confocal microscope(LSCM)has been used in the field of morphological research of acupuncture and moxibustion for more than 20 years.It is one of the important tools for revealing the structure of acupoints and the effect of acupuncture from histological perspective.With the help of technical advantages of LSCM,the quality of morphological research of acupuncture and moxibustion has been greatly improved,helping us gain a deeper understanding about the structure of acupoints and meridians as well as histochemical changes induced by acupuncture/moxibustion intervention.In order to promote the application of LSCM in acupuncture and moxibustion,we simply reviewed some recent studies in this field and combined them with our experience,trying to provide some technical suggestions.We expect that the technique of LSCM could be integrated into more experiments in acupuncture medicine to provide more powerful morphological evidence for exploring the underlying mechanisms of acupuncture and moxibution therapies.
Laser scanning confocal microscope(LSCM)has been used in the field of morphological research of acupuncture and moxibustion for more than 20 years.It is one of the important tools for revealing the structure of acupoints and the effect of acupuncture from histological perspective.With the help of technical advantages of LSCM,the quality of morphological research of acupuncture and moxibustion has been greatly improved,helping us gain a deeper understanding about the structure of acupoints and meridians as well as histochemical changes induced by acupuncture/moxibustion intervention.In order to promote the application of LSCM in acupuncture and moxibustion,we simply reviewed some recent studies in this field and combined them with our experience,trying to provide some technical suggestions.We expect that the technique of LSCM could be integrated into more experiments in acupuncture medicine to provide more powerful morphological evidence for exploring the underlying mechanisms of acupuncture and moxibution therapies.
引文
[1]逄树龙,蔡振宇.激光扫描共聚焦显微镜在医学研究中的应用[J].现代生物医学进展,2009,9(13):2579-2580.
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[3]陶之理,张祖萍,王良培,等.足三里穴区感觉神经元的节段性分布(HRP法的研究)[J].针刺研究,1983,8(2):136-140.
[4]蒋瑾,白万柱,张璐,等.大鼠“太溪”穴区与神经系统的节段和区域相关性研究——辣根过氧化物酶法[J].针刺研究,2010,35(1):27-31.
[5]朱新龙,白万柱,吴富东,等.大鼠“承山”穴的神经解剖学特征——霍乱毒素亚单位B结合荧光素488法[J].针刺研究,2010,35(6):433-437.
[6]崔晶晶,朱新龙,吉长福,等.大鼠“京骨”和“大钟”原络配穴的神经解剖学基础——霍乱毒素亚单位B结合荧光素488和594双标法[J].针刺研究,2011,36(4):262-267.
[7]CUI J J,HA L J,ZHU X L,et al.Neuroanatomical basis for acupuncture point PC 8in the rat:neural tracing study with cholera toxin subunit B[J].Acupunct Med,2013,31(4):389-394.
[8]WU M L,CUI J J,XU D S,et al.Neuroanatomical characteristics of deep and superficial needling using LI 11 as an example[J].Acupunct Med,2015,33(6):472-477.
[9]CUI J J,ZHU X L,SHI H,et al.The expression of calcitonin gene-related peptide on the neurons associated Zusanli(ST 36)in rats[J].Chin J Integr Med,2015,21(8):630-634.
[10]WU M L,XU D S,BAI W Z,et al.Local cutaneous nerve terminal and mast cell responses to manual acupuncture in acupoint LI 4area of the rats[J].J Chem Neuroanat,2015,68:14-21.
[11]吴美玲.伤害性神经肽与肥大细胞在针刺穴位中的应答[D].北京:中国中医科学院,2016.
[12]何伟,吴美玲,景向红,等.穴位的本态:穴位组织细胞化学的动态变化[J].中国针灸,2015,35(11):1181-1186.
[13]黄光英,郑翠红,张明敏.针刺对大鼠“足三里”穴缝隙连接蛋白Cx 43表达的影响[J].中国针灸,2005,25(8):565-568.
[14]HE W,WANG X Y,SHI H,et al.Cutaneous neurogenic inflammation in the sensitized acupoints induced by gastric mucosal injury in rats[J].BMC Complement Altern Med,2017,17(1):141.
[15]李强.“凤汉学说”沉浮的启示[J].中国针灸,2011,31(3):263-268.
[16]林栋,黄晓真,庄婉玉,等.从“凤汉系统”到“原始管道系统”——对经穴实质研究的思考[J].中国针灸,2017,37(1):95-101.
[17]LEE B C,KIM S,SOH K S.Novel anatomic structures in the brain and spinal cord of rabbit that may belong to the Bonghan system of potential acupuncture meridians[J].J Acupunct Meridian Stud,2008,1(1):29-35.
[18]LEE B C,BAE K H,JHON G J,et al.Bonghan system as mesenchymal stem cell niches and pathways of macrophages in adipose tissues[J].J Acupunct Meridian Stud,2009,2(1):79-82.
[19]LEE B C,KIM H B,SUNG B,et al.Network of endocardial vessels[J].Cardiology,2011,118(1):1-7.
[20]LIM C J,LEE S Y,RYU P D.Identification of primo-vascular system in abdominal subcutaneous tissue layer of rats[J].Evid Based Complement Alternat Med,2015,2015:751937.
[21]WANG X Y,SHI H,CUI J J,et al.Preliminary research of relationship between acute peritonitis and celiac primo vessels[J].Evid Based Complement Alternat Med,2013,2013:569161.
[22]TIAN Y Y,JING X H,GUO S G,et al.Study on the formation of novel threadlike structure through intravenous injection of heparin in rats and refined observation in minipigs[J].Evid Based Complement Alternat Med,2013,2013:731518.
[23]吉长福,王晓宇,石宏,等.原始管道系统(PVs)与针灸经络关系的研究[J].上海针灸杂志,2013,32(10):795-799.
[24]LOAIZA L A,YAMAGUCHI S,ITO M,et al.Electro-acupuncture stimulation to muscle afferents in anesthetized rats modulates the blood flow to the knee joint through autonomic reflexes and nitric oxide[J].Auton Neurosci,2002,97(2):103-109.
[25]罗燕,许能贵,易玮,等.电针对局灶性脑缺血大鼠大脑皮层缺血灶周围区星形胶质细胞的影响[J].针刺研究,2009,34(2):101-105.
[26]许能贵,易玮,赖新生,等.电针对局灶性脑缺血大鼠脑细胞内Ca2+含量的影响[J].中国中西医结合杂志,2002,22(4):295-297.
[27]杨帆,徐国龙,杨永清,等.电针对戊四唑点燃型癫痫大鼠脑电及海马神经元内Ca2+含量的影响[J].针刺研究,2009,34(3):163-166.
[28]ZHAO H,TIAN Z Z,CHEN B Y.An important role of corticotropin-releasing hormone in electroacupuncture normalizing the subnormal function of hypothalamus-pituitary-ovary axis in ovariectomized rats[J].Neurosci Lett,2003,349(1):25-28.
[29]ZHU Y Z,ZENG Y J.Electroacupuncture protected pyramidal cells in hippocampal CA 1region of vascular dementia rats by inhibiting the expression of p 53and Noxa[J].CNS Neurosci Ther,2011,17(6):599-604.
[30]CUI J J,HA L J,ZHU X L,et al.Specificity of sensory and motor neurons associated with BL 40and GB 30in the rat:a dual fluorescent labeling study[J].Evid Based Complement Alternat Med,2013,2013:643403.
[31]张坤,徐东升,崔晶晶,等.P物质在大鼠“三阴交”穴区相关感觉神经元和神经纤维中的表达[J].针刺研究,2015,40(6):449-454.
[32]张知云,徐东升,崔晶晶,等.P物质阳性表达神经纤维在大鼠心包经五输穴局部皮肤组织中的分布[J].针刺研究,2016,41(4):298-302.
[33]徐东升,佘琛,王佳,等.大鼠“太冲”穴区皮肤组织中血管和神经纤维的显微结构特征[J].针刺研究,2016,41(6):486-491.
[34]ZHANG W J,XU D S,CUI J J,et al.Anterograde and retrograde tracing with high molecular weight biotinylated dextran amine through thalamocortical and corticothalamic pathways[J].Microsc Res Tech,2017,80(2):260-266.
[2]李瑞午,景向红,蔡虹.激光共焦扫描显微镜在针灸经络研究中的应用(推荐一种现代科学研究手段)[J].针刺研究,1995,20(2):76-78.
[3]陶之理,张祖萍,王良培,等.足三里穴区感觉神经元的节段性分布(HRP法的研究)[J].针刺研究,1983,8(2):136-140.
[4]蒋瑾,白万柱,张璐,等.大鼠“太溪”穴区与神经系统的节段和区域相关性研究——辣根过氧化物酶法[J].针刺研究,2010,35(1):27-31.
[5]朱新龙,白万柱,吴富东,等.大鼠“承山”穴的神经解剖学特征——霍乱毒素亚单位B结合荧光素488法[J].针刺研究,2010,35(6):433-437.
[6]崔晶晶,朱新龙,吉长福,等.大鼠“京骨”和“大钟”原络配穴的神经解剖学基础——霍乱毒素亚单位B结合荧光素488和594双标法[J].针刺研究,2011,36(4):262-267.
[7]CUI J J,HA L J,ZHU X L,et al.Neuroanatomical basis for acupuncture point PC 8in the rat:neural tracing study with cholera toxin subunit B[J].Acupunct Med,2013,31(4):389-394.
[8]WU M L,CUI J J,XU D S,et al.Neuroanatomical characteristics of deep and superficial needling using LI 11 as an example[J].Acupunct Med,2015,33(6):472-477.
[9]CUI J J,ZHU X L,SHI H,et al.The expression of calcitonin gene-related peptide on the neurons associated Zusanli(ST 36)in rats[J].Chin J Integr Med,2015,21(8):630-634.
[10]WU M L,XU D S,BAI W Z,et al.Local cutaneous nerve terminal and mast cell responses to manual acupuncture in acupoint LI 4area of the rats[J].J Chem Neuroanat,2015,68:14-21.
[11]吴美玲.伤害性神经肽与肥大细胞在针刺穴位中的应答[D].北京:中国中医科学院,2016.
[12]何伟,吴美玲,景向红,等.穴位的本态:穴位组织细胞化学的动态变化[J].中国针灸,2015,35(11):1181-1186.
[13]黄光英,郑翠红,张明敏.针刺对大鼠“足三里”穴缝隙连接蛋白Cx 43表达的影响[J].中国针灸,2005,25(8):565-568.
[14]HE W,WANG X Y,SHI H,et al.Cutaneous neurogenic inflammation in the sensitized acupoints induced by gastric mucosal injury in rats[J].BMC Complement Altern Med,2017,17(1):141.
[15]李强.“凤汉学说”沉浮的启示[J].中国针灸,2011,31(3):263-268.
[16]林栋,黄晓真,庄婉玉,等.从“凤汉系统”到“原始管道系统”——对经穴实质研究的思考[J].中国针灸,2017,37(1):95-101.
[17]LEE B C,KIM S,SOH K S.Novel anatomic structures in the brain and spinal cord of rabbit that may belong to the Bonghan system of potential acupuncture meridians[J].J Acupunct Meridian Stud,2008,1(1):29-35.
[18]LEE B C,BAE K H,JHON G J,et al.Bonghan system as mesenchymal stem cell niches and pathways of macrophages in adipose tissues[J].J Acupunct Meridian Stud,2009,2(1):79-82.
[19]LEE B C,KIM H B,SUNG B,et al.Network of endocardial vessels[J].Cardiology,2011,118(1):1-7.
[20]LIM C J,LEE S Y,RYU P D.Identification of primo-vascular system in abdominal subcutaneous tissue layer of rats[J].Evid Based Complement Alternat Med,2015,2015:751937.
[21]WANG X Y,SHI H,CUI J J,et al.Preliminary research of relationship between acute peritonitis and celiac primo vessels[J].Evid Based Complement Alternat Med,2013,2013:569161.
[22]TIAN Y Y,JING X H,GUO S G,et al.Study on the formation of novel threadlike structure through intravenous injection of heparin in rats and refined observation in minipigs[J].Evid Based Complement Alternat Med,2013,2013:731518.
[23]吉长福,王晓宇,石宏,等.原始管道系统(PVs)与针灸经络关系的研究[J].上海针灸杂志,2013,32(10):795-799.
[24]LOAIZA L A,YAMAGUCHI S,ITO M,et al.Electro-acupuncture stimulation to muscle afferents in anesthetized rats modulates the blood flow to the knee joint through autonomic reflexes and nitric oxide[J].Auton Neurosci,2002,97(2):103-109.
[25]罗燕,许能贵,易玮,等.电针对局灶性脑缺血大鼠大脑皮层缺血灶周围区星形胶质细胞的影响[J].针刺研究,2009,34(2):101-105.
[26]许能贵,易玮,赖新生,等.电针对局灶性脑缺血大鼠脑细胞内Ca2+含量的影响[J].中国中西医结合杂志,2002,22(4):295-297.
[27]杨帆,徐国龙,杨永清,等.电针对戊四唑点燃型癫痫大鼠脑电及海马神经元内Ca2+含量的影响[J].针刺研究,2009,34(3):163-166.
[28]ZHAO H,TIAN Z Z,CHEN B Y.An important role of corticotropin-releasing hormone in electroacupuncture normalizing the subnormal function of hypothalamus-pituitary-ovary axis in ovariectomized rats[J].Neurosci Lett,2003,349(1):25-28.
[29]ZHU Y Z,ZENG Y J.Electroacupuncture protected pyramidal cells in hippocampal CA 1region of vascular dementia rats by inhibiting the expression of p 53and Noxa[J].CNS Neurosci Ther,2011,17(6):599-604.
[30]CUI J J,HA L J,ZHU X L,et al.Specificity of sensory and motor neurons associated with BL 40and GB 30in the rat:a dual fluorescent labeling study[J].Evid Based Complement Alternat Med,2013,2013:643403.
[31]张坤,徐东升,崔晶晶,等.P物质在大鼠“三阴交”穴区相关感觉神经元和神经纤维中的表达[J].针刺研究,2015,40(6):449-454.
[32]张知云,徐东升,崔晶晶,等.P物质阳性表达神经纤维在大鼠心包经五输穴局部皮肤组织中的分布[J].针刺研究,2016,41(4):298-302.
[33]徐东升,佘琛,王佳,等.大鼠“太冲”穴区皮肤组织中血管和神经纤维的显微结构特征[J].针刺研究,2016,41(6):486-491.
[34]ZHANG W J,XU D S,CUI J J,et al.Anterograde and retrograde tracing with high molecular weight biotinylated dextran amine through thalamocortical and corticothalamic pathways[J].Microsc Res Tech,2017,80(2):260-266.