药学学报, 2020, 55(7): 1614-1621
引用本文:
马欢欢, 朱晓悫, 郭文燕, 郝杰, 靳保花, 张炜. 布比卡因抑制谷氨酸诱导大鼠海马星形胶质细胞内钙升高的机制[J]. 药学学报, 2020, 55(7): 1614-1621.
MA Huan-huan, ZHU Xiao-que, GUO Wen-yan, HAO Jie, JIN Bao-hua, ZHANG Wei. Mechanisms of bupivacaine inhibition of glutamate-induced intracellular calcium elevation in primary cultured rat hippocampal astrocytes[J]. Acta Pharmaceutica Sinica, 2020, 55(7): 1614-1621.

布比卡因抑制谷氨酸诱导大鼠海马星形胶质细胞内钙升高的机制
马欢欢, 朱晓悫, 郭文燕, 郝杰, 靳保花, 张炜
河北医科大学中西医结合研究所, 河北 石家庄 050017
摘要:
本文以原代培养的大鼠海马星形胶质细胞为研究对象,探讨布比卡因(bupivacaine)抑制谷氨酸诱导的海马星形胶质细胞内钙升高的机制。运用免疫荧光技术确定代谢型谷氨酸受体(mGluR5受体)在神经元和星形胶质细胞上的表达情况;利用钙离子成像技术,观察布比卡因和mGluR5受体拮抗剂2-甲基-6-(苯乙炔)吡啶盐酸盐[2-methyl-6-(2-phenylethynyl)-pyridine,MPEP]对谷氨酸诱导的大鼠海马神经元和星形胶质细胞胞内游离钙(intracellular free Ca2+concentration,[Ca2+]i)变化的影响;观察布比卡因对mGluRs(mGluR1/5)受体激动剂(RS)-3,5-二羟基苯基甘氨酸[(RS)-3,5-dihydroxyphenylglycine,DHPG]及mGluR5受体特异性激动剂2-氯-5羟苯基甘氨酸钠盐[(RS)-2-chloro-5-hydroxyphenylglycine sodium salt,CHPG]诱导的大鼠海马神经元和星形胶质细胞[Ca2+]i变化的影响。动物实验已获得河北医科大学实验动物伦理委员会批准。结果显示,神经元和星形胶质细胞上mGluR5受体表达丰富;300 μmol·L-1布比卡因可抑制1 mmol·L-1谷氨酸诱导的星形胶质细胞[Ca2+]i的升高(P<0.01);10 μmol·L-1 MPEP完全抑制了1 mmol·L-1谷氨酸诱导的星形胶质细胞[Ca2+]i的升高(P<0.01),而对神经元[Ca2+]i升高反应的抑制率仅为10%~20%;300 μmol·L-1布比卡因可完全抑制50 μmol·L-1 DHPG介导的星形胶质细胞[Ca2+]i升高的反应(P<0.01);布比卡因浓度依赖性地抑制CHPG诱导的星形胶质细胞[Ca2+]i的升高,IC50约为100 μmol·L-1。以上研究结果表明,布比卡因通过作用于星形胶质细胞上的mGluR5受体抑制谷氨酸介导的海马星形胶质细胞[Ca2+]i升高的反应。
关键词:    布比卡因      mGluR5受体      谷氨酸      海马星形胶质细胞     
Mechanisms of bupivacaine inhibition of glutamate-induced intracellular calcium elevation in primary cultured rat hippocampal astrocytes
MA Huan-huan, ZHU Xiao-que, GUO Wen-yan, HAO Jie, JIN Bao-hua, ZHANG Wei
Institute of Chinese Integrative Medicine, Hebei Medical University, Shijiazhuang 050017, China
Abstract:
This research investigated the mechanism by which bupivacaine inhibits glutamate-induced intracellular free Ca2+ increases in primary cultured hippocampal astrocytes. Immunofluorescence was used to demonstrate the expression of metabotropic glutamate receptor (mGluR5 receptor) on neurons and astrocytes. Calcium imaging was used to measure the alteration of intracellular free Ca2+ ([Ca2+]i) in primary cultured rat hippocampal neurons and astrocytes. The animal experiments were approved by the Animal Experiments Ethical Committee of Hebei Medical University. The results showed that mGluR5 receptor was abundantly expressed in the primary cultured rat neurons and astrocytes. Bupivacaine (300 μmol·L-1) significantly inhibited 1 mmol·L-1 glutamate-induced[Ca2+]i increase in astrocytes (P<0.01). 2-Methyl-6-(2-phenylethynyl)-pyridine (MPEP) (10 μmol·L-1) completely abolished the increase of[Ca2+]i induced by 1 mmol·L-1 glutamate in the astrocytes (P<0.01), while the inhibitory effect on neurons was only 10%-20%. Bupivacaine (300 μmol·L-1) completely inhibited the[Ca2+]i increase induced by mGluR5 receptor agonists (RS)-3,5-dihydroxyphenylglycine (DHPG) (50 μmol·L-1) and (RS)-2-chloro-5-hydroxyphenylglycine sodium salt (CHPG) (1 mmol·L-1) in astrocytes (P<0.01). In addition, bupivacaine inhibited the CHPG-induced[Ca2+]i increase in a dose-dependent manner in astrocytes with an IC50 of 100 μmol·L-1. The results from this study indicate that bupivacaine inhibits glutamate-induced[Ca2+]i elevation by acting on the mGluR5 receptor in primary cultured hippocampal astrocytes.
Key words:    bupivacaine    mGluR5 receptor    glutamate    astrocyte   
收稿日期: 2020-05-26
DOI: 10.16438/j.0513-4870.2020-0842
基金项目: 国家自然科学基金资助项目(81872848);科技部重大新药创制项目(2018ZX09711001-004-003);京津冀基础研究项目(H2018206641);河北省三三三人才项目(A201701010068).
通讯作者: 张炜,Tel:13933877472,E-mail:weizhang@hebmu.edu.cn
Email: weizhang@hebmu.edu.cn
相关功能
PDF(582KB) Free
打印本文
0
作者相关文章
马欢欢  在本刊中的所有文章
朱晓悫  在本刊中的所有文章
郭文燕  在本刊中的所有文章
郝杰  在本刊中的所有文章
靳保花  在本刊中的所有文章
张炜  在本刊中的所有文章

参考文献:
[1] Ruetsch YA, Boni T, Borgeat A. From cocaine to ropivacaine:the history of local anesthetic drugs[J]. Curr Top Med Chem, 2001, 1:175-182.
[2] Kato J, Konishi J, Yoshida H, et al. Cauda equina syndrome following combined spinal and epidural anesthesia:a case report[J]. Can J Anaesth, 2011, 58:638-641.
[3] Killeen T, Kamat A, Walsh D, et al. Severe adhesive arachnoiditis resulting in progressive paraplegia following obstetric spinal anaesthesia:a case report and review[J]. Anaesthesia, 2012, 67:1386-1394.
[4] Karaca O, Pinar HU, Dogan R. Total spinal block, bupivacaine toxicity or else under epidural anaesthesia?[J]. Agri, 2017, 29:149-150.
[5] Moore PA, Hersh EV. Local anesthetics:pharmacology and toxicity[J]. Dent Clin North Am, 2010, 54:587-599.
[6] Malet A, Faure MO, Deletage N, et al. The comparative cytotoxic effects of different local anesthetics on a human neuroblastoma cell line[J]. Anesth Analg, 2015, 120:589-596.
[7] Crosby KM, Murphy-Royal C, Wilson SA, et al. Cholecystokinin switches the plasticity of GABA synapses in the dorsomedial hypothalamus via astrocytic ATP release[J]. J Neurosci, 2018, 38:8515-8525.
[8] Umpierre AD, West PJ, White JA, et al. Conditional knock-out of mGluR5 from astrocytes during epilepsy development impairs high-frequency glutamate uptake[J]. J Neurosci, 2019, 39:727-742.
[9] Xing P, Ma K, Li L, et al. The protection effect and mechanism of hyperbaric oxygen therapy in rat brain with traumatic injury[J]. Acta Cir Bras, 2018, 33:341-353.
[10] Kono M, Kakegawa W, Yoshida K, et al. Interneuronal NMDA receptors regulate long-term depression and motor learning in the cerebellum[J]. J Physiol, 2019, 597:903-920.
[11] Gasparini CF, Sutherland HG, Haupt LM, et al. Genetic analysis of GRIA2 and GRIA4 genes in migraine[J]. Headache, 2014, 54:303-312.
[12] Vanzulli I, Butt AM. mGluR5 protect astrocytes from ischemic damage in postnatal CNS white matter[J]. Cell Calcium, 2015, 58:423-430.
[13] Johnson ME, Saenz JA, DaSilva AD, et al. Effect of local anesthetic on neuronal cytoplasmic calcium and plasma membrane lysis (necrosis) in a cell culture model[J]. Anesthesiology, 2002, 97:1466-1476.
[14] Arai Y, Kondo T, Tanabe K, et al. Enhancement of hyperthermia-induced apoptosis by local anesthetics on human histiocytic lymphoma U937 cells[J]. J Biol Chem, 2002, 277:18986-18993.
[15] Xing Y, Zhang N, Zhang W, et al. Bupivacaine indirectly potentiates glutamate-induced intracellular calcium signaling in rat hippocampal neurons by impairing mitochondrial function in cocultured astrocytes[J]. Anesthesiology, 2018, 128:539-554.
[16] Zhang N, Xiong WW, Xing Y, et al. Culture method of rat fetal hippocampal neurons and astrocytes[J]. Acta Neuropharmacologica (神经药理学报), 2017, 7:24-28.
[17] Albright GA. Cardiac arrest following regional anesthesia with etidocaine or bupivacaine[J]. Anesthesiology, 1979, 51:285-287.
[18] Liu P, Feldman HS, Covino BM, et al. Acute cardiovascular toxicity of intravenous amide local anesthetics in anesthetized ventilated dogs[J]. Anesth Analg, 1982, 61:317-322.
[19] Abe T, Sugihara H, Nawa H, et al. Molecular characterization of a novel metabotropic glutamate receptor mGluR5 coupled to inositol phosphate/Ca2+ signal transduction[J]. J Biol Chem, 1992, 267:13361-13368.
[20] Wang JW, Wang HD. Research progress of metabolic glutamate receptor 5 in glial cells[J]. Chin J Neurosurg Dis Res (中华神经外科疾病研究杂志), 2013, 12:470-471.