目的：通过对吗啡戒断、耐受小鼠进行电针和穴位埋线治疗,观察小鼠海马和脊髓中N-甲基-D-天冬氨酸(N-methyl-D-aspartate, NMDA)受体和胆囊收缩素(cholecystokinin, CCK)的表达水平,探讨电针和埋线对吗啡戒断后戒断、耐受小鼠调整作用的差异。方法将56只雄性C57BL/6J小鼠随机分为戒断空白组、戒断模型组、戒断埋线组、戒断电针组和耐受空白组、耐受模型组、耐受埋线组、耐受电针组,每组7只。吗啡戒断模型根据“7 d递增成瘾法”皮下注射盐酸吗啡注射液,戒断空白组在相同时间点注射等量生理盐水,戒断电针组小鼠每次注射盐酸吗啡注射液15 min 后,使用韩式穴位神经刺激仪(HANS-200)电针刺激双侧肾俞穴；戒断埋线组于注射盐酸吗啡注射液15 min 后,将0.5 cm铬肠线埋入双侧肾俞穴内。第7天早上10点,腹腔注射盐酸纳洛酮注射液催瘾(4 mg/kg),并观察小鼠戒断反应。根据阿片依赖戒断症状柳田知司测评量表评分,并以酶联免疫吸附剂测定法(Elisa)检测海马和脊髓NMDA受体和CCK含量。按10 mg/kg皮下注射吗啡建立吗啡耐受模型。耐受空白组在相同的时间注射10 mL/kg的生理盐水,耐受埋线组于建立模型第1天在肾俞穴埋线治疗,耐受电针组在建立模型第1天起电针肾俞穴。经7 d治疗后,以Elisa方法检测海马和脊髓的 NMDA受体和CCK含量。结果戒断模型组海马NR2B和CCK表达与戒断空白组比较,差异均具有统计学意义(P＜0.05)。戒断电针组海马NR2B表达与戒断模型组比较,差异有统计学意义(P＜0.05)。戒断埋线组和戒断电针组海马 CCK 表达与戒断模型组比较,差异均具有统计学意义(P＜0.05)。戒断模型组小鼠脊髓NR2A、NR2B、CCK表达与戒断空白组比较,差异均具有统计学意义(P＜0.05)。戒断电针组脊髓NR2A、NR2B表达与戒断模型组比较,差异均具有统计学意义(P＜0.05)。耐受模型组海马NR1、NR2B、CCK表达与耐受空白组比较,差异均具有统计学意义(P＜0.05)。耐受埋线组海马CCK表达与耐受模型组比较,差异具有统计学意义(P＜0.05)。耐受电针组海马NR1表达与耐受模型组和耐受埋线组比较,差异均具有统计学意义(P＜0.05)。耐受埋线组和耐受电针组脊髓CCK表达与耐受模型组比较,差异均具有统计学意义(P＜0.05)。结论肾俞穴穴位埋线及电针治疗对减轻吗啡耐受及戒断均有治疗效果,且电针治疗优于穴位埋线。%Objective To observe expression levels of N-methyl-D-aspartate (NMDA) receptor and cholecystokinin (CCK) in the hippocampus and spinal cord in morphine withdrawal or tolerance mice treated by electroacupuncture or catgut embedding and explore the difference between the regulating effects of electroacupuncture and catgut embedding on morphine withdrawal and tolerance.Methods Fifty-six male C57BL/6J mice were randomly allocated to withdrawal control, withdrawal model, withdrawal catgut embedding and withdrawal electroacupuncture groups, and tolerance control, tolerance model, tolerance catgut embedding and tolerance electroacupuncture groups, 7 mice in each group. A model of morphine withdrawal was made by subcutaneous injection of morphine hydrochloride using 7-day increasing addiction method. The withdrawal control group was injected with an equal volume of normal saline at the same time points. In the withdrawal electroacupuncture group, electroacupuncture at bilateral points Shenshu was performed using a Han’s acupoint nerve stimulation device (HANS-200) at 15 min after an injection of morphine hydrochloride. In the withdrawal catgut embedding group, 0.5 cm chromic catgut was embedded in bilateral points Shenshu at 15 min after an injection of morphine hydrochloride. Addiction was promoted by intraperitoneal injection of naloxone 4 mg/kg at 10 o’clock on the seventh day’s morning and Withdrawal reactions were observed in the mice. The score was recorded using the Ryuta Tomoji opioid withdrawal symptoms evaluation scale. NMDA receptor and CCK contents in the hippocampus and spinal cord were measured by enzyme-linked immunosorbent assay (ELISA). A model of morphine tolerance was made by subcutaneous injection of morphine 10 mg/kg. The tolerance control group was injected with tolerance normal saline 10 ml/kg at the same time. In the tolerance catgut embedding group, catgut was embedded in point Shenshu at the first day after model making. In the tolerance electroacupuncture group, point Shenshu was given electroacupuncture at the first day after model making. After seven days of treatment, NMDA receptor and CCK contents in the hippocampus and spinal cord were measured by ELISA.Results There were statistically significant differences in hippocampal NR2B and CCK expressions between the withdrawal model and withdrawal control groups (P<0.05). There was a statistically significant difference in hippocampal NR2B expression between the withdrawal electroacupuncture and withdrawal model groups (P<0.05). There was a statistically significant difference in hippocampal CCK expression between the withdrawal catgut embedding or withdrawal electroacupuncture group and the withdrawal model group (P<0.05). There were statistically significant differences in spinal cord NR2A, NR2B and CCK expressions between the withdrawal model and withdrawal control groups (P<0.05). There were statistically significant differences in spinal cord NR2A and NR2B expressions between the withdrawal electroacupuncture and withdrawal model groups (P<0.05). There were statistically significant differences in hippocampal NR2A, NR2B and CCK expressions between the tolerance model and tolerance control groups (P<0.05). There was a statistically significant difference in hippocampal CCK expression between the tolerance catgut embedding and tolerance model groups (P<0.05). There was a statistically significant difference in hippocampal NR1 expression between the tolerance electroacupuncture group and the tolerance model or tolerance catgut embedding group (P<0.05). There was a statistically significant difference in spinal cord CCK expression between the tolerance catgut embedding or withdrawal electroacupuncture group and the tolerance model group (P<0.05).Conclusions Both catgut embedding and electroacupuncture at point Shenshu have a reducing effect on morphine tolerance and withdrawal. The therapeutic effect of electroacupuncture is better than that of catgut embedding.
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