胍丁胺对吗啡耐受小鼠蓝斑核组织中μ-阿片受体和自噬相关蛋白的作用机制  

Effects of agmatine onμ-opioid receptors and autophagy related proteins in the locus coeruleus of morphine-tolerant mice

作　　者：兰岚 陆巍 LAN Lan;LU Wei(Department of Pain Diagnosis and Treatment,School of Anesthesiology,Guizhou Medical University,Guiyang 550004,Guizhou,China)

机构地区：[1]贵州医科大学麻醉学院疼痛诊疗学教研室,贵州贵阳550004 [2]贵州医科大学

出　　处：《贵州医科大学学报》2023年第1期63-69,共7页Journal of Guizhou Medical University

基　　金：贵州省卫生健康委科学技术基金项目(gzwkj2023-395)。

摘　　要：目的探讨胍丁胺(AG)对吗啡(Mor)耐受小鼠蓝斑核(LC)组织中μ-阿片受体(MOR)和自噬相关蛋白的作用机制。方法40只成年健康雄性C57BL/6J小鼠随机分为生理盐水(NS)组、Mor组、AG-Mor组及AG组,皮下注射给药法建立小鼠慢性Mor耐受模型,采用温水甩尾法测定各组小鼠给药前及给药第1~7天的甩尾潜伏期(TFL);造模结束后麻醉处死小鼠、取脑组织,采用免疫荧光法、Western blot及荧光定量聚合酶链式反应(q-PCR)检测各组小鼠LC组织中MOR的表达,采用Western blot法检测各组小鼠LC组织中自噬蛋白微管相关蛋白1轻链3(LC3)、家蚕隔离体蛋白1(P62)的表达。结果与Mor组比较,AG-Mor组小鼠TFL下降趋势较缓慢,差异有统计学意义(P<0.05);与Mor组比较,AG-Mor组小鼠LC组织中MOR蛋白表达水平升高、P62含量明显下降,差异有统计学意义(P<0.05);与NS组比较,Mor组小鼠LC组织中LC3-Ⅱ/LC3-Ⅰ比值与P62含量均出现不同程度的升高,差异有统计学意义(P<0.05)。结论AG抗Mor耐受形成的机制可能与增加Mor耐受小鼠LC组织中MOR蛋白表达量、降低P62含量有关。Objective To investigate the possible effects and mechanism of agmatine(AG)onμ-opioid receptors(MOR)and autophagy related protein in locus coeruleus(LC)tissues of morphine(Mor)tolerant mice.Methods Forty healthy adult male C57BL/6J mice were randomly divided into four groups:normal saline(NS)group,Mor group,AG-Mor group,and AG group(n=10 in each group).Chronic Mor tolerant models were established by subcutaneous injection.The tail flapping latency(TFL)was measured by warm water to observe the analgesic effect.After the modeling,the subject mice were anesthetized and killed,and the brain tissue samples were taken.The expression of MOR in LC was detected by immunofluorescence(IF),Western blot,and q-PCR.Western blot was used to detect the expression of autophagy related proteins microtubuleassociated protein 1 light chain 3(LC3)and sequestosome-1(P62)in LC of mice.Results Compared with Mor group,TFL decreased more slowly in AG-Mor group,and the difference was statistically significant(P<0.05).Compared with Mor group,the expression level of MOR protein in LC region of AG-Mor group increased while the content of P62 decreased significantly,with statistical significance(P<0.05).Compared with NS group,the ratio of LC3-Ⅱ/LC3-Ⅰ,and the content of P62 in LC region of Mor group increased to varying degrees,with statistical significance(P<0.05).Conclusion AG can increase the expression ofμ-opioid receptor protein and decrease the content of P62 in LC of Mortolerant mice.

关 键 词：胍丁胺 自噬 吗啡耐受 Μ阿片受体 蓝斑核 自噬蛋白微管相关蛋白1轻链3 家蚕隔离体蛋白1 

分 类 号：R614[医药卫生—麻醉学] R441.1[医药卫生—外科学]