独活和独活香豆素对帕金森病模型大鼠脑黑质CHOP和Caspase12表达的影响

Effect of Radix Angelicae Pubescentis and Coumarins on Expressions of CHOP and Caspase12 in Substantia Nigra of Rats with Parkinson’s Disease

目的:通过观察独活及独活香豆素对乳胞素(Lactacystin)诱导的帕金森病(PD)模型大鼠脑黑质CHOP和Caspase12表达水平的影响,探讨其与内质网应激的关系。方法:利用乳胞素(Lactacystin)脑黑质致密部注射建立PD大鼠模型,于第7、14、21、28天进行阿扑吗啡(APO)检测,根据大鼠行为学异常程度,应用分层随机法将模型复制的大鼠随机分为模型对照组、吐温80组、独活香豆素组、独活颗粒剂组、独活复方颗粒剂组、塞来昔布组、美多芭组七组。另设假手术组和正常对照组,假手术组脑黑质致密部注射等量生理盐水。正常对照组、假手术组、模型对照组用生理盐水灌胃,其他组大鼠同时间予以等体积相应药物灌胃,共10周。给药后第14、28、42、56、70天分别进行APO检测,观察记录其旋转行为异常、旋转方向、旋转圈数。在进行最后一次APO检测后的次日,过量麻醉处死大鼠,断头,取左侧脑黑质,采用RT-PCR、Western-blot法,对大鼠脑黑质CHOP、Caspase12 mRNA及蛋白表达水平进行检测。结果:应用Lactacystin诱导的PD模型大鼠脑黑质CHOP和Caspase12的表达明显上调;予以药物干预后,独活香豆素、独活颗粒剂、独活复方颗粒剂、美多芭、塞来昔布组大鼠脑黑质CHOP和Caspase12的表达均有下降,以独活香豆素和或独活颗粒剂组最为显著。结论:独活和独活香豆素可能是通过抑制内质网应激对PD起到保护作用。

Objective: To address the relationship of endoplasmic reticulum stress and the protective effects of Radix Angelicae Pubescentis and its active constituents coumarins on Parkinson’s disease(PD), we observed the expression levels of CHOP and Caspase12 in the substantia nigra of rats with PD induced by lactacystin. Method: PD model was established in rats by injecting lactacystin into the dense part of substantia nigra and the level of apomorphine(APO) was measured on the 7th, 14th, 21th and 28th day after lactacystin injection. By means of stratified random method, rats with PD were divided into 7 groups according the abnormal degrees of their behavior: model control group, Tween-80 group, Angelica pubescens coumarin group, Angelica pubescens granule group, Compound granule of Angelica pubescens group, Celecoxib group and Medopa group. Besides, two more groups were set up: the sham group that the rats were injected the same amount of normal saline into the substantia nigra and the normal control group. Normal control group, sham group and model control group were given intragastric administration of normal saline. The other groups were given equal volume of drugs for 10 weeks at the same time. APO level was measured on the 14th, 28th, 42th, 56th and 70th day after drug administration. The abnormal rotation behaviors were recorded, such as rotation direction and rotation number. Rats were sacrificed on the next day after the last measurement of APO and the left substantia nigras were collected and the expressions of CHOP and Caspase12 were determined by RT-PCR and Western-blot. Results: The expressions of CHOP and Caspase12 were upregulated in the substantia nigra of PD rats induced by lactacystin, however, their expressions were all decreased after the intervention of Angelica pubescens coumarin, Angelica pubescens granule, Compound granule of Angelica pubescens, Celecoxib and Medopa. Among them, Angelica pubescens coumarin and Angelica pubescens granule are the most significant. Conclusion: Radix Angelicae Pubescentis and coumarins from Radix Angelicae may play a protective role in PD by inhibiting endoplasmic recticulun stress.