深部脑刺激内侧前额皮层对大鼠海洛因觅药行为的干预作用
发布时间：2018-09-03 20:57
【摘要】：目的：深部脑刺激（deep brain stimulation，DBS)临床上已用于帕金森病难治性强迫症癫痫和抑郁症等精神疾病的治疗。和其他电刺激技术或脑手术相比，DBS具有可逆性、微创性和安全性高等优点。近几年，有实验研究和临床的初步观察提示DBS具有治疗药物依赖及防复吸潜力。但有关DBS治疗海洛因复吸的基础和临床研究刚起步，治疗作用、最佳刺激参数和合适靶核团的选择以及作用途径尚待进一步系统研究。本课题旨在利用大鼠海洛因自身给药模型，选用内侧前额皮层（medial prefrontal cortex，mPFC）的两个亚区（背和腹内侧前额皮层）进行DBS处理，观察DBS对大鼠海洛因觅药行为消退和恢复的影响，并初步探讨其可能的分子生物学作用机制。 实验一、慢性高频或低频刺激背内侧前额皮层（dorsal medial prefrontalcortex，dmPFC）对大鼠海洛因觅药行为消退和恢复的影响 方法：SD成年雄性大鼠经颈静脉插管和埋置电极手术后恢复7天，所有大鼠均进行每天4h的海洛因固定比率（FR1）自身给药训练，连续训练14h，建立海洛因自身给药模型。27只大鼠随机分为假刺激对照组（Sham组）高频刺激处理组（H-DBS组）和低频刺激处理组（L-DBS组）(n=9)，然后进行环境消退训练，每天2h。每天消退训练前，刺激处理组大鼠在dmPFC脑区给予1h高频（频率130HZ，脉宽100μs，电流0.2mA）或低频（频率10HZ，脉宽100μs，，电流0.2mA）刺激处理，共连续刺激10d，然后在最后一次消退训练结束24h后进行线索诱导的海洛因觅药行为测试。假刺激对照组刺激电流为0mA，其余同刺激处理组。 结果：已建立稳定的海洛因自身给药大鼠，在随后的第1-10天的消退训练中，H-DBS组和L-DBS组有效鼻触数较Sham对照组有效鼻触数均无显著性差异（P0.05）；在线索诱导的海洛因觅药行为恢复测试中，单因素方差分析发现：H-DBS组有效鼻触数显著低于Sham对照组有效鼻触数（F(2,24)=5.304, P 0.05），而L-DBS组有效鼻触数与Sham对照组相比无显著性差异（P0.05）。 实验二慢性高频或低频刺激腹内侧前额皮层(ventral medial prefrontalcortex，vmPFC)对大鼠海洛因觅药行为消退和恢复的影响 方法：海洛因自身给药海洛因自身给药模型建立、消退训练、DBS电极埋置和刺激参数等同实验一。所有海洛因自身给药大鼠随机分为vmPFC脑区高频刺激处理组（H-DBS组）、低频刺激处理组（L-DBS组）和假刺激对照组（Sham组）(n=9)。 结果：与Sham对照组相比，消退训练第1-3天，H-DBS组有效鼻触数有增高趋势，但无显著性差异，随着消退训练天数的继续，消退训练第4-7天，H-DBS组有效鼻触数显著高于Sham对照组有效鼻触数（P0.05），而在消退训练第8-10天中，与Sham对照组相比，H-DBS组有效鼻触数均无显著性差异；而L-DBS组在消退训练第1-10天中，其有效鼻触数较Sham对照组相比均无显著性差异（P0.05）；在线索诱导的海洛因觅药行为恢复测试中，单因素方差分析发现：与Sham对照组相比，H-DBS组有效鼻触数显著增加（F(2,24)=8.489, P 0.05），而L-DBS组有效鼻触数无显著性差异（P0.05） 实验三慢性高频刺激dmPFC对大鼠伏隔核核部（nucleus accumbensshell, NAc shell）和壳部（nucleus accumbens core, NAc core）p-CREBp-ERK和p-Akt表达的影响 方法：海洛因自身给药海洛因自身给药模型建立、消退训练、DBS电极埋置、刺激参数和线索诱导的觅药行为恢复测试等同实验一，所有海洛因自身给药大鼠随机分为dmPFC脑区高频刺激处理组（H-DBS组）和假刺激对照组（Sham组）(n=9)。线索诱导的觅药行为测试结束，立即断头取脑：采用Western blot方法检测NAc shell和NAc core中p-CREBp-ERK和p-AKt的表达水平变化；运用免疫组化方法，观察高频刺激dmPFC对NAc shell和NAccore中p-CREB蛋白阳性表达的影响。 结果：Western blot检测发现：与Sham对照组相比，H-DBS组NAc core中p-CREB表达显著升高（t(6)=9.342，P0.01），p-ERK与p-AKt表达均显著减少（t(6)=13.347，P0.01；t(6)=11.678，P0.01），但H-DBS组NAc shell中p-CREB和p-ERK均无显著性变化（P0.05），而p-AKt表达显著减少（t(6)=3.863，P0.05）；同时，在p-CREB免疫组化实验中发现，与Sham对照组相比， H-DBS组NAc core中p-CREB阳性细胞数显著增加（t(6)=12.107，P0.05），而NAcshell中p-CREB阳性细胞数无显著变化（P0.05）。 结论 高频刺激dmPFC能抑制线索诱导的海洛因觅药行为恢复，其作用可能与NAc中磷酸化CREBERK和AKt蛋白的表达改变有关。高频刺激vmPFC可促进线索诱导的海洛因觅药行为的恢复，并对大鼠觅药行为消退反应有一定的抑制作用；但是，低频刺激dmPFC或vmPFC对大鼠觅药行为消退和线索诱导的觅药行为的恢复均没有明显的干预作用。
[Abstract]:OBJECTIVE: Deep brain stimulation (DBS) has been used clinically for the treatment of Parkinson's disease, refractory obsessive-compulsive disorder, epilepsy and depression. Compared with other electrical stimulation techniques or brain surgery, DBS has the advantages of reversibility, minimally invasiveness and high safety. However, the basic and clinical studies on DBS in the treatment of heroin relapse have just begun, and the therapeutic effects, the optimal stimulation parameters, the selection of appropriate target nuclei and the pathway of action need to be further systematically studied. Two subareas (dorsal and ventromedial prefrontal cortex) of L prefrontal cortex (mPFC) were treated with DBS to observe the effects of DBS on the regression and recovery of heroin seeking behavior in rats, and to explore the possible molecular biological mechanism.
Experiment 1. Effects of chronic high or low frequency stimulation of dorsal medial prefrontal cortex (dmPFC) on the regression and recovery of heroin seeking behavior in rats
METHODS: SD adult male rats recovered 7 days after jugular vein catheterization and implantation of electrodes. All rats were trained for 4 hours a day for self-administration of heroin (FR1). Heroin self-administration model was established after 14 hours of continuous training. 27 rats were randomly divided into sham group, high-frequency stimulation group (H-DBS group) and low-frequency stimulation group (H-DBS group). Frequency stimulation group (L-DBS group) (n=9), and then environmental regression training, 2 hours a day. Before regression training, stimulation group rats were given 1 hour high frequency (frequency 130HZ, pulse width 100 mus, CURRENT 0.2 mA) or low frequency (frequency 10HZ, pulse width 100 mus, CURRENT 0.2 mA) stimulation in the brain region of dmPFC for 10 days, and then the last regression training session. The cue-induced heroin seeking behavior was tested 24 hours later. The stimulation current of the sham stimulation control group was 0 mA, and the rest of the sham stimulation group was the same as the stimulation treatment group.
RESULTS: Stable heroin self-administered rats were established, and there was no significant difference between H-DBS group and L-DBS group (P 0.05) in effective nasal contact number compared with Sham control group in the following 1-10 days of regression training. The number of effective nasal contacts in L-DBS group was significantly lower than that in Sham control group (F(2,24)=5.304,P 0.05), but there was no significant difference between L-DBS group and Sham control group (P 0.05).
Effect of chronic high or low frequency stimulation of ventral medial prefrontal cortex (vmPFC) on the regression and recovery of heroin seeking behavior in rats
METHODS: Heroin self-administration model was established, regression training, DBS electrode implantation and stimulation parameters were the same as experiment 1. All heroin self-administration rats were randomly divided into three groups: high-frequency stimulation group (H-DBS group), low-frequency stimulation group (L-DBS group) and sham control group (n=9).
Results: Compared with Sham control group, the number of effective nasal contacts in H-DBS group increased from 1 to 3 days of regression training, but there was no significant difference. With the continuation of regression training days, the number of effective nasal contacts in H-DBS group was significantly higher than that in Sham control group on 4-7 days of regression training (P 0.05), but in 8-10 days of regression training, the number of effective nasal contacts in H-DBS group was significantly higher than that in Sham control group. There was no significant difference in the number of effective nasal contacts between DBS group and Sham control group, but there was no significant difference in the number of effective nasal contacts between L-DBS group and Sham control group (P 0.05). (F (2,24) =8.489, P 0.05), but there was no significant difference in the number of effective nasal contacts in the L-DBS group (P0.05).
Effect of chronic high frequency stimulation of dmPFC on the expression of p-CREB? P-ERK and p-Akt in nucleus accumbens (NAc shell) and nucleus accumbens core (NAc core) of rats
METHODS: Heroin self-administration model was established, regression training, DBS electrode implantation, stimulus parameters and cue-induced drug-seeking behavior recovery test were the same as experiment 1. All heroin self-administration rats were randomly divided into dmPFC high-frequency stimulation group (H-DBS group) and sham control group (n=9). The expression of p-CREB?P-ERK and p-AKt in NAc shell and NAc core was detected by Western blot, and the effect of high frequency stimulation of dmPFC on the expression of p-CREB protein in NAc shell and NAc core was observed by immunohistochemistry.
Results: Western blot showed that the expression of p-CREB in NAc core of H-DBS group was significantly higher than that of Sham control group (t (6) = 9.342, P 0.01), and the expression of p-ERK and p-AKt were significantly decreased (t (6) = 13.347, P 0.01; t (6) = 11.678, P 0.01), but the expression of p-CREB and p-ERK in NAc shell of H-DBS group was not significantly changed (P 0.05), but p-AKt (6) = 3.863, P 0.863, P 0.01). At the same time, compared with Sham control group, the number of p-CREB positive cells in NAc core of H-DBS group increased significantly (t(6) = 12.107, P 0.05), while the number of p-CREB positive cells in NAc Shell did not change significantly (P 0.05).
conclusion
High frequency stimulation of dmPFC can inhibit the recovery of clue-induced heroin seeking behavior, which may be related to the changes of phosphorylated CREB?ERK and AKt protein expression in NAc. There was no significant effect of mPFC or vmPFC on the regression of drug-seeking behavior and the recovery of clue-induced drug-seeking behavior in rats.
【学位授予单位】：宁波大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R749.64
本文编号：2221144
[Abstract]:OBJECTIVE: Deep brain stimulation (DBS) has been used clinically for the treatment of Parkinson's disease, refractory obsessive-compulsive disorder, epilepsy and depression. Compared with other electrical stimulation techniques or brain surgery, DBS has the advantages of reversibility, minimally invasiveness and high safety. However, the basic and clinical studies on DBS in the treatment of heroin relapse have just begun, and the therapeutic effects, the optimal stimulation parameters, the selection of appropriate target nuclei and the pathway of action need to be further systematically studied. Two subareas (dorsal and ventromedial prefrontal cortex) of L prefrontal cortex (mPFC) were treated with DBS to observe the effects of DBS on the regression and recovery of heroin seeking behavior in rats, and to explore the possible molecular biological mechanism.
Experiment 1. Effects of chronic high or low frequency stimulation of dorsal medial prefrontal cortex (dmPFC) on the regression and recovery of heroin seeking behavior in rats
METHODS: SD adult male rats recovered 7 days after jugular vein catheterization and implantation of electrodes. All rats were trained for 4 hours a day for self-administration of heroin (FR1). Heroin self-administration model was established after 14 hours of continuous training. 27 rats were randomly divided into sham group, high-frequency stimulation group (H-DBS group) and low-frequency stimulation group (H-DBS group). Frequency stimulation group (L-DBS group) (n=9), and then environmental regression training, 2 hours a day. Before regression training, stimulation group rats were given 1 hour high frequency (frequency 130HZ, pulse width 100 mus, CURRENT 0.2 mA) or low frequency (frequency 10HZ, pulse width 100 mus, CURRENT 0.2 mA) stimulation in the brain region of dmPFC for 10 days, and then the last regression training session. The cue-induced heroin seeking behavior was tested 24 hours later. The stimulation current of the sham stimulation control group was 0 mA, and the rest of the sham stimulation group was the same as the stimulation treatment group.
RESULTS: Stable heroin self-administered rats were established, and there was no significant difference between H-DBS group and L-DBS group (P 0.05) in effective nasal contact number compared with Sham control group in the following 1-10 days of regression training. The number of effective nasal contacts in L-DBS group was significantly lower than that in Sham control group (F(2,24)=5.304,P 0.05), but there was no significant difference between L-DBS group and Sham control group (P 0.05).
Effect of chronic high or low frequency stimulation of ventral medial prefrontal cortex (vmPFC) on the regression and recovery of heroin seeking behavior in rats
METHODS: Heroin self-administration model was established, regression training, DBS electrode implantation and stimulation parameters were the same as experiment 1. All heroin self-administration rats were randomly divided into three groups: high-frequency stimulation group (H-DBS group), low-frequency stimulation group (L-DBS group) and sham control group (n=9).
Results: Compared with Sham control group, the number of effective nasal contacts in H-DBS group increased from 1 to 3 days of regression training, but there was no significant difference. With the continuation of regression training days, the number of effective nasal contacts in H-DBS group was significantly higher than that in Sham control group on 4-7 days of regression training (P 0.05), but in 8-10 days of regression training, the number of effective nasal contacts in H-DBS group was significantly higher than that in Sham control group. There was no significant difference in the number of effective nasal contacts between DBS group and Sham control group, but there was no significant difference in the number of effective nasal contacts between L-DBS group and Sham control group (P 0.05). (F (2,24) =8.489, P 0.05), but there was no significant difference in the number of effective nasal contacts in the L-DBS group (P0.05).
Effect of chronic high frequency stimulation of dmPFC on the expression of p-CREB? P-ERK and p-Akt in nucleus accumbens (NAc shell) and nucleus accumbens core (NAc core) of rats
METHODS: Heroin self-administration model was established, regression training, DBS electrode implantation, stimulus parameters and cue-induced drug-seeking behavior recovery test were the same as experiment 1. All heroin self-administration rats were randomly divided into dmPFC high-frequency stimulation group (H-DBS group) and sham control group (n=9). The expression of p-CREB?P-ERK and p-AKt in NAc shell and NAc core was detected by Western blot, and the effect of high frequency stimulation of dmPFC on the expression of p-CREB protein in NAc shell and NAc core was observed by immunohistochemistry.
Results: Western blot showed that the expression of p-CREB in NAc core of H-DBS group was significantly higher than that of Sham control group (t (6) = 9.342, P 0.01), and the expression of p-ERK and p-AKt were significantly decreased (t (6) = 13.347, P 0.01; t (6) = 11.678, P 0.01), but the expression of p-CREB and p-ERK in NAc shell of H-DBS group was not significantly changed (P 0.05), but p-AKt (6) = 3.863, P 0.863, P 0.01). At the same time, compared with Sham control group, the number of p-CREB positive cells in NAc core of H-DBS group increased significantly (t(6) = 12.107, P 0.05), while the number of p-CREB positive cells in NAc Shell did not change significantly (P 0.05).
conclusion
High frequency stimulation of dmPFC can inhibit the recovery of clue-induced heroin seeking behavior, which may be related to the changes of phosphorylated CREB?ERK and AKt protein expression in NAc. There was no significant effect of mPFC or vmPFC on the regression of drug-seeking behavior and the recovery of clue-induced drug-seeking behavior in rats.
【学位授予单位】：宁波大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R749.64
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相关期刊论文 前1条
1 徐纪文,王桂松,周洪语,田鑫,王祥瑞,应隽,朱玫娟,张新凯,虞一萍,江基尧,罗其中;深部脑刺激戒断阿片类药物精神依赖1例临床报道(附3个月随访结果)[J];立体定向和功能性神经外科杂志;2005年03期
本文编号：2221144
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