TCM  >> Vol. 6 No. 2 (May 2017)

    滋肾平颤颗粒对异动症大鼠行为学及纹状体多巴胺D1受体活性和基因表达的影响
    Effect of Zishenpingchan Granule on Neurobehavioral Manifestations and the Activity and Gene Expression of Striatal Dopamine D1 Receptors of Rats with Levodopa-Induced Dyskinesias

  • 全文下载: PDF(1757KB) HTML   XML   PP.118-126   DOI: 10.12677/TCM.2017.62020  
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作者:  

叶青,周洁,袁晓蕾,袁灿兴:上海中医药大学附属龙华医院脑病科;
高东雯:上海中医药大学附属龙华医院超声科

关键词:
异动症大鼠滋肾平颤颗粒行为学多巴胺D1受体直接通路LID Rats Zishenpingchan Granule Dopamine D1 Receptors Direct-Pathway

摘要:

目的:观察中药对异动症(levodopa-induced dyskinesias, LID)大鼠行为学和纹状体多巴胺D1受体活性及mRNA表达的影响。方法:6-OHDA构建(Parkinson’s disease, PD)大鼠模型,左旋多巴治疗4周制作LID大鼠模型,将成功建立的LID大鼠,随机分为模型组、中药干预组,另取相同数量的正常大鼠作为正常对照组,治疗四周后观察中药对LID大鼠异常不自主运动(abnormal involuntary move-ment, AIM)评分、剂峰旋转次数的影响;测定大鼠纹状体多巴胺D1受体的最大结合容量(maximum binding capacity, Bmax)和平衡解离常数(equilibrium dissociation constant, KD);运用实时定量PCR检测多巴胺D1受体mRNA表达。结果:模型组大鼠随着左旋多巴治疗时间的延长,AIM评分及剂峰旋转次数进行性升高;与模型组比较,中药干预后可明显减少异动症大鼠的AIM积分及旋转次数(P < 0.05)。纹状体多巴胺D1受体活性结果显示:两组大鼠与正常对照组比较Bmax升高(P < 0.01),KD值降低(P < 0.05),受体活性明显增高,中药治疗可使Bmax降低,KD值升高(P < 0.01),明显降低D1受体活性;荧光实时定量PCR结果显示模型组大鼠多巴胺D1受体mRNA表达明显上调(P < 0.01);中药可以显著下调多巴胺D1受体mRNA的表达(P < 0.05)。结论:多巴胺D1受体介导的直接通路过度激活可能是异动症产生的主要机制。中药主要是通过下调多巴胺D1受体活性及基因表达从而有效缓解异动症症状。

Objective: To observe the effects of Traditional Chinese Medicine on neurobehavioral manifestations and the activity and mRNA expression of striatal dopamine D1 receptors of rats with levodo-pa-induced dyskinesias (LID). Methods: The rat model of Parkinson’s disease (PD) was established by 6-OHDA; then, the model of LID in rat was produced by injecting levodopa (LD) and benserazide for 4 weeks. The rats were divided into normal control group, LID model group and TCM interven-tion group. After 4 weeks of treatment, the effect of TCM on abnormal involuntary movement (AIM) score of rats with LID, peak dose rotation and efficacy time of levodopa (LD) were observed. The maximum binding capacity (Bmax) and equilibrium dissociation constant (KD) of striatal dopamine D1 receptors of rats were determined. The mRNA expression of dopamine D1 receptors was assayed by using real-time fluorescent quantitative PCR (FQ-PCR). Result: With the extension of LD treat-ment time, the AIM score and peak dose rotation of rats in the LID model group increased progres-sively. Comparing to the LD model group, TCM intervention could obviously reduce the AIM score and peak dose rotation of rats with LID (P < 0.05). The research of D1 receptor activity showed that level of Bmax increased (P < 0.01) and level of KD reduced (P < 0.01) in the LID model group and the TCM intervention group in comparison to those in the normal control group. The activity of D1 receptor increased obviously. TCM treatment could decrease the Bmax level, increase the KD level (P < 0.01) and lower the activity of D1 receptor. The results of FQ-PCR analysis revealed that the gene expression of D1 receptor was up-regulated in the LID model group (P < 0.01). TCM treatment can decrease over-expressed D1 receptor expression (P < 0.05). Conclusion: Over-activation in di-rect-pathway mediated by dopamine D1 receptor may be the main mechanism of LID. TCM effec-tively relieved LID symptoms through regulating the gene expression and activity of dopamine D1 receptors.

文章引用:
叶青, 高东雯, 周洁, 袁晓蕾, 袁灿兴. 滋肾平颤颗粒对异动症大鼠行为学及纹状体多巴胺D1受体活性和基因表达的影响[J]. 中医学, 2017, 6(2): 118-126. https://doi.org/10.12677/TCM.2017.62020

参考文献

[1] Zesiewicz, T.A., Sullivan, K.L. and Hauser, R.A. (2007) Levodopa-Induced Dyskinesia in Parkinson’s Disease: Epide-miology, Etiology, and Treatment. Current Neurology and Neuroscience Reports, 7, 302-310.
https://doi.org/10.1007/s11910-007-0046-y
[2] Encarnacion, E.V. and Hauser, R.A. (2008) Levodopa-Induced Dyskinesias in Parkinson’s Disease: Etiology, Impact on Quality of Life and Treatments. European Neurology, 60, 57-66.
https://doi.org/10.1159/000131893
[3] Cao, X.B., Guan, Q., Xu, Y., et al. (2006) Mechanism of Over-Activation in Direct Pathway Mediated by Dopamine D1 Receptor in Rats with Levodopa-Induced Dyskinesia. Neuroscience Bulletin, 22, 159-164.
[4] Fabbrini, G., Brotchie, J.M., Grandas, F., et al. (2007) Levodopa-Induced Dyskinesias. Movement Disorders, 22, 1379- 1389.
https://doi.org/10.1002/mds.21475
[5] 巴茂文, 刘振国, 孔敏, 等. 左旋多巴诱发帕金森病大鼠异动症模型的建立和评价[J]. 上海交通大学学报(医学版), 2006, 26(7): 810-812.
[6] 包新民, 舒思云. 大鼠脑立体定位图谱[M]. 北京: 人民卫生出版社, 1991: 53.
[7] Carman, L.S., Gage, G.H. and Shults, C.W. (1991) Partial Lesion of the Substantia Nigra: Relation between Extent of 1esion and Rota-tional Behavior. Brain Research, 553, 275-283.
[8] 曹学兵, 孙圣刚, 徐岩, 等. 左旋多巴治疗帕金森病诱发异动症的实验研究[J]. 中华医学杂志, 2004, 84(6): 505-507.
[9] 李丹, 黄议腺, 王晓军, 等. 左旋多巴诱发异动症6-羟基多巴帕金森病大鼠模型的建立及行为学评价[J]. 苏州大学学报: 医学版, 2010, 30(2): 236-239.
[10] 叶青, 周洁, 袁灿兴. 滋肾平颤颗粒对异动症大鼠行为学及黑质纹状体DAT和VMAT2基因表达的影响[J]. 江苏中医药, 2014, 46(4): 74-76.
[11] 张旺明, 蔡颖谦, 徐如祥, 等. 帕金森病大鼠模型尾壳核多巴胺D2受体活性变化研究[J]. 中华神经外科疾病研究杂志, 2002, 1(3): 266-268.
[12] Obeso, J.A., Rodriguez-Oroz, M.C., Rodriguez, M., et al. (2000) Pathophysiology of Levodopa-Induced Dyskinesias in Parkinson’s Disease: Problems with the Current Model. Discussion. Annals of Neurology, 47, S22-S34.
[13] Utsumi, H., Okuma, Y., Kano, O., et al. (2013) Evaluation of the Efficacy of Pramipexole for Treating Levodopa-In- duced Dyskinesia in Patients with Parkinson’s Disease. Internal Medicine, 52, 325-332.
[14] Brotchie, J.M., Lee, J. and Venderova, K. (2005) Levodopa-Induced Dyskinesia in Parkin-son, s Disease. Journal of Neural Transmission, 112, 359-391.
https://doi.org/10.1007/s00702-004-0251-7
[15] Mela, F., Marti, M., Bido, S., et al. (2012) In Vivo Evidence for a Dif-ferential Contribution of Striatal and Nigral D1 and D2 Receptors to L-DOPA Induced Dyskinesia and the Accompany-ing Surge of Nigral Amino Acid Levels. Neurobiology of Disease, 45, 573-582.
[16] Santini, E., Valjent, E. and Fisone, G. (2008) Parkinson’s Disease: Levodopa-Induced Dyskinesia and Signal Transduction. The FEBS Journal, 275, 1392-1399.
https://doi.org/10.1111/j.1742-4658.2008.06296.x
[17] 周洁, 叶青, 袁灿兴. 熟地平颤汤治疗帕金森氏病的随机对照研究[J]. 中华中医药学刊, 2014, 32(6): 1395-1397.
[18] Ye, Q., Yuan, X.-L., Zhou, J., Yuan, C. and Yang, X.-M. (2014) Effect of Zishenpingchan Granule on Neurobehavioral Manifestations and the Activity and Gene Expression of Striatal Dopamine D1 and D2 Receptors of Rats with Levodopa-Induced Dyskinesias. Evi-dence-Based Complementary and Alternative Medicine, 2014, Article ID: 342506.
[19] Ye, Q., Yuan, X.-L., He, J., et al. (2016) Anti-Apoptotic Effect of Shudipingchan Granule in the Substantia Nigra of Rat Models of Parkinson’s Dis-ease. Neural Regeneration Research, 11, 1625-1632.
https://doi.org/10.4103/1673-5374.193242