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  

Mechanisms of bupivacaine inhibition of glutamate-induced intracellular calcium elevation in primary cultured rat hippocampal astrocytes

作　　者：马欢欢 朱晓悫 郭文燕 郝杰 靳保花 张炜 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)

机构地区：[1]河北医科大学中西医结合研究所,河北石家庄050017

出　　处：《药学学报》2020年第7期1614-1621,共8页Acta Pharmaceutica Sinica

基　　金：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)。

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;300μmol·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);10μmol·L-1MPEP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%;300μmol·L-1布比卡因可完全抑制50μmol·L-1DHPG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100μmol·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。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(300μmol·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)(10μmol·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(300μmol·L-1)completely inhibited the[Ca2+]i increase induced by mGluR5 receptor agonists(RS)-3,5-dihydroxyphenylglycine(DHPG)(50μmol·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 100μmol·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.

关 键 词：布比卡因 mGluR5受体 谷氨酸 海马星形胶质细胞 

分 类 号：R964[医药卫生—药理学]