熟地黄-山茱萸药干预缺血性卒中模型大鼠脑组织的蛋白组学研究

doi:10.5246/jcps.2023.01.003

中国药学(英文版) ›› 2023, Vol. 32 ›› Issue (1): 32-44.DOI: 10.5246/jcps.2023.01.003

熟地黄-山茱萸药干预缺血性卒中模型大鼠脑组织的蛋白组学研究

李昕¹, 商燕¹, 齐梦欣¹, 胡国恒²^,^*()

1. 湖南中医药大学, 湖南长沙 410208
2. 湖南中医药大学第一附属医院, 湖南长沙 410007

收稿日期:2022-09-20 修回日期:2022-10-15 接受日期:2022-11-26 出版日期:2023-01-31 发布日期:2023-01-31
通讯作者: 胡国恒
作者简介:
+ Tel.: +86-13974888299, E-mail: huguoheng@hnucm.edu.cn
基金资助:
National Natural Science Foundation of China (Grant No. 81774208).

Proteomic study of Rehmannia glutinosa and Cornus officinalis herbal pair on the brain tissue of rats with ischemic stroke

Xin Li¹, Yan Shang¹, Mengxin Qi¹, Guoheng Hu²^,^*()

1 Hunan University of Chinese Medicine, Changsha 410208, Hunan, China
2 The First Affiliated Hospital of Hunan University of Chinese Medicine, Changsha 410007, Hunan, China

Received:2022-09-20 Revised:2022-10-15 Accepted:2022-11-26 Online:2023-01-31 Published:2023-01-31
Contact: Guoheng Hu

摘要/Abstract

摘要：

鉴于中医药"补肾填精"治法对缺血性中风有着良好的治疗效果, 本研究选取熟地黄-山茱萸作为代表性药物组合探讨该治法对缺血性中风的治疗作用。使用SD大鼠建立MCAO模型并分为模型组与治疗组两组, 另设假手术组, 每组10只。治疗组每日以熟地黄-山茱萸药对汤剂灌胃, 其余组以等体积蒸馏水灌胃, 干预7天后取梗死半球脑组织, 进行TTC染色评估脑梗死区域体积, 并应用串联质谱标签技术检测各组大鼠梗死区域脑组织蛋白组学变化。结果显示熟地黄-山茱萸药能有效保护缺血脑组织, 梗死体积较模型组明显降低, 蛋白组学检测质量可靠, 各组之间蛋白组表达谱有显著差异。其中治疗组与模型组之间差异表达的蛋白共993个, 其中上调756个, 下调237个, 这些差异蛋白的功能主要涉及多种酶的活性调节及多种分子结合功能的改变, 相关的信号通路功能主要与酮体的合成和降解、多种神经突触功能、磷脂酰肌醇信号系统、促进细胞新陈代谢等有关。这些分子功能与通路可能是该药对干预缺血性卒中的靶点, 可为下一步研究提供参考。

关键词: 熟地黄, 山茱萸, 缺血性卒中, 蛋白组学, 补肾填精

Abstract:

Since the traditional Chinese medicine (TCM) "tonifying kidney and filling essence" treatment has an excellent therapeutic effect on ischemic stroke, Rehmannia glutinosa and Cornus officinalis were selected as the representative herbal combination to explore the effect of this therapeutic method on ischemic stroke. SD rats were used to establish the MCAO model and divided into two groups, including the model group and treatment group. Another sham operation group was set up, with 10 rats in each group. The treatment group was treated daily with Rehmannia Cornus Decoction by gavage, while the sham group and model group were treated with the same volume of distilled water by gavage. Brain tissue of the infarcted hemisphere was collected after 7 d of continuous intervention, then TTC staining was performed to evaluate the cerebral infarction volume, and the proteomic changes in the infarcted region of rats were detected by tandem mass tag spectrometry technology. The results showed that Rehmannia glutinosa and Cornus officinalis herb pair could effectively protect ischemic brain tissue and showed reliable proteomic assay quality and significant differences in proteomic expression profiles between the groups. A total of 993 proteins were differentially expressed between the treatment and model groups, of which 756 were up-regulated, and 237 were down-regulated. The functions of these differential proteins were mainly involved in regulating the activity of various enzymes and the changes in the binding functions of various molecules. The signaling pathways involved were mainly related to the synthesis and degradation of ketone, various synaptic functions, and phosphatidylinositol signaling system. These molecular functions and pathways might be the targets of this herbal pair for the intervention of ischemic stroke and provide a reference for further research.

Key words: Rehmannia glutinosa, Cornus officinalis, Ischemic stroke, Proteomics, Tonifying kidney and essence

Supporting:

李昕, 商燕, 齐梦欣, 胡国恒. 熟地黄-山茱萸药干预缺血性卒中模型大鼠脑组织的蛋白组学研究[J]. 中国药学(英文版), 2023, 32(1): 32-44.

Xin Li, Yan Shang, Mengxin Qi, Guoheng Hu. Proteomic study of Rehmannia glutinosa and Cornus officinalis herbal pair on the brain tissue of rats with ischemic stroke[J]. Journal of Chinese Pharmaceutical Sciences, 2023, 32(1): 32-44.

图/表 5

Figure 1. Representative of TTC-stained brain slices (A) and infarct volume quantitation (B). Error bars represent as mean ± SD. *P = 0.0068 vs. the model group using the Student’s t-test. ND: not detected.

Figure 2. Coomassie blue staining of each sample and TIC/BPC plot of a random fraction. (A) Each band shows one mixed sample. MG: model group, SG: sham operation group, and TG: treatment group. (B) In the TIC/BPC, the signal intensities in each segment were generally consistent, the chromatographic peaks were sharp, and the curves were smooth, indicating that the chromatography was effective and stable.

Figure 3. Volcano plot of differential proteins among groups. (A) Model group vs. sham group; (B) treatment group vs. model group; (C) treatment group vs. sham group. FC > 1.2 or < 1/1.2.

Figure 4. Go enrichment analysis involving results from the top five differential protein quantities. (A) Model group vs. sham group; (B) treatment group vs. model group.

Figure 5. Bubble plot of KEGG pathway enrichment of differential proteins between groups. (A) Model group vs. sham group; (B) treatment group vs. model group; (C) treatment group vs. sham group.

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熟地黄-山茱萸药干预缺血性卒中模型大鼠脑组织的蛋白组学研究

Proteomic study of Rehmannia glutinosa and Cornus officinalis herbal pair on the brain tissue of rats with ischemic stroke

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