电针对骨癌痛-吗啡耐受大鼠蓝斑核μ阿片受体表达的干预

发布时间：2018-05-04 05:07

本文选题：骨癌痛 + 吗啡耐受　；参考：《中国针灸》2017年05期

【摘要】：目的:观察电针对骨癌痛-吗啡耐受(cancer pain and morphine tolerance)大鼠痛行为的影响及部分作用机制。方法:将42只健康雌性SD大鼠完全随机分为5组:假手术组(7只)、骨癌痛组(8只)、吗啡耐受组(9只)、电针组(9只)和假电针组(9只)。假手术组于左胫骨髓腔内注射无菌磷酸盐缓冲液,其余4组大鼠均于左胫骨髓腔内注射MRMT-1乳腺癌细胞以制备骨癌痛模型。假手术组及骨癌痛组术后均不予干预。吗啡耐受组、电针组及假电针组于术后第11d对骨癌痛制备成功大鼠行盐酸吗啡注射液腹腔注射,每12h注射1次,连续注射11d诱导骨癌痛-吗啡耐受模型。此模型建立后第1d起,吗啡耐受组每12h(早上9:00,晚上9:00)行吗啡注射,连续7d;电针组、假电针组均于早上9:00行吗啡注射,30min后分别给予电针(2Hz/100Hz)和假电针(仅刺入皮下,破皮即可)治疗,穴取双侧"足三里"和"昆仑",每次30min,每日1次,连续治疗7d。分别于癌细胞接种前1d,术后第6d、8d、10d,吗啡注射第1d、5d、9d、11d的30min后及电针治疗第1d、3d、5d、7d的30min后检测各组大鼠患侧机械缩足阈(paw withdrawal threshold,PWT)的变化。于吗啡注射第11d,采用HE染色检测假手术组、骨癌痛组、吗啡耐受组大鼠(每组随机选2只)胫骨组织形态学变化;电针治疗第7d采用荧光免疫组织化学法观察各组大鼠(每组随机选4只)蓝斑核μ阿片受体(μ-opioid receptor,MOR)阳性细胞表达情况。结果:癌细胞接种后第10d,28只骨癌痛造模成功大鼠(骨癌痛组8只、吗啡耐受组8只、电针组6只、假电针组6只)PWT较7只假手术组大鼠明显下降(P0.01)。吗啡注射第1d,吗啡耐受组、电针组及假电针组大鼠PWT均明显高于骨癌痛组(均P0.01);吗啡连续注射第11d,吗啡耐受组、电针组、假电针组大鼠PWT与骨癌痛组比较差异均无统计学意义(均P0.05)。吗啡注射第11d,骨癌痛组、吗啡耐受组大鼠胫骨上1/3处均可见癌细胞致肿块,且胫骨髓腔内布满MRMT-1癌细胞;假手术组大鼠胫骨未见异常变化。电针治疗第1d、3d、5d、7d,骨癌痛组、吗啡耐受组和假电针组大鼠患侧PWT均明显低于电针组(均P0.01)。电针治疗第7d,骨癌痛组、吗啡耐受组、电针组、假电针组蓝斑核MOR阳性表达均低于假手术组(P0.01,P0.05),且骨癌痛组、吗啡耐受组和假电针组均低于电针组(均P0.01)。结论:电针可提高骨癌痛-吗啡耐受大鼠的机械痛阈,改善模型大鼠痛觉异常;该效应可能与电针提高大鼠蓝斑核MOR阳性细胞表达有关。
[Abstract]:Objective: to observe the effect and mechanism of electroacupuncture on pain behavior of pain and morphine tolerance rats. Methods: 42 healthy female SD rats were randomly divided into 5 groups: sham operation group (n = 7), bone cancer pain group (n = 8), morphine tolerance group (n = 9), electroacupuncture group (n = 9) and pseudoelectroacupuncture group (n = 9). In sham-operated group, aseptic phosphate buffer solution was injected into the left tibial medullary cavity, and MRMT-1 breast cancer cells were injected into the left tibial medullary cavity in the other four groups to establish the model of bone cancer pain. There was no intervention after operation in the sham operation group and the bone cancer pain group. Rats in morphine tolerance group, electroacupuncture group and pseudoelectroacupuncture group were injected morphine hydrochloride intraperitoneally every 12 hours on the 11th day after operation. The model of pain and morphine tolerance of bone cancer was induced by continuous injection for 11 days. One day after the establishment of the model, morphine was injected into the morphine tolerance group every 12 hours (9: 00 in the morning, 9: 00 in the evening) for 7 consecutive days, while in the electric acupuncture group, the sham electroacupuncture group was given electroacupuncture 2Hz / 100Hz2 and pseudoelectroacupuncture (only subcutaneously) at 9:00 after morphine injection for 30 minutes. For 7 days, both sides of Zusanli and Kunlun were treated once a day for 30 mins. The changes of paw withdrawal thresholdPWTs were detected at 1 day before inoculation of cancer cells, 8 days after operation, 11 days after morphine injection for 5 days and 9 days after treatment with electroacupuncture for 3 days and 5 days and 7 days after 30min. At the 11th day of morphine injection, the histomorphology of tibia was detected by HE staining in sham-operation group, bone cancer pain group and morphine tolerance group (2 rats in each group). On the 7th day of electroacupuncture treatment, the expression of 渭 -opioid receptor (渭 -opioid receptor) positive cells in the locus coeruleus of rats (4 rats in each group) was observed by fluorescence immunohistochemical method. Results: on the 10th day after inoculation of cancer cells, 28 successful models of bone cancer pain were established in 28 rats (8 in bone cancer pain group, 8 in morphine tolerance group, 6 in electroacupuncture group, 6 in sham electroacupuncture group and 6 in sham operation group). The PWT in sham operation group was significantly lower than that in sham operation group (P 0.01). On the 1st day of morphine injection, the PWT of rats in morphine tolerance group, electroacupuncture group and pseudoacupuncture group were significantly higher than those in bone cancer pain group (all P 0.01), morphine tolerance group, electroacupuncture group, morphine tolerance group and electroacupuncture group on the 11th day of continuous morphine injection. There was no significant difference in PWT between sham electroacupuncture group and bone cancer pain group (P 0.05). On the 11th day of morphine injection, tumor cells were found in the bone cancer pain group and morphine tolerance group, and MRMT-1 cancer cells were found in the medullary cavity of the tibia, but there was no abnormal change in the tibia in the sham operation group. The PWT of the affected side of the rats in the pain group, morphine tolerance group and pseudo-electroacupuncture group were significantly lower than that in the electroacupuncture group (all P 0.01). On the 7th day after electroacupuncture treatment, the positive expression of MOR in bone-cancer pain group, morphine tolerance group, electroacupuncture group and pseudo-electroacupuncture group were all lower than that in sham operation group (P 0.01), and the expression of MOR in bone cancer pain group, morphine tolerance group and pseudoelectroacupuncture group were lower than that in electroacupuncture group (all P 0.01). Conclusion: electroacupuncture can increase the mechanical pain threshold of rats with pain and morphine tolerance of bone cancer and ameliorate the abnormal pain perception in the model rats. This effect may be related to the increase of MOR positive cells expression in the nucleus coeruleus of rats.
【作者单位】：浙江中医药大学第三临床医学院针灸神经生物学实验室;嘉兴学院附属第一医院针灸科;
【基金】：国家自然科学基金资助项目:81102643 浙江省自然科学基金资助项目:LY 14H270016,LQ 15H270003,LY 16H270017 中国博士后基金面上项目:2014 M550334 浙江省医药卫生科研项目:2014KYA 162
【分类号】：R245

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