Pharmacoproteomics reveals the mechanism of Chinese dragon's blood in regulating the RSK/TSC2/mTOR/ribosome pathway in alleviation of DSS-induced acute ulcerative colitis
Pharmacoproteomics reveals the mechanism of Chinese dragon's blood in regulating the RSK/TSC2/mTOR/ribosome pathway in alleviation of DSS-induced acute ulcerative colitis
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DOI：
10.1016/j.jep.2020.113221
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发表时间：
2020-12-05
影响因子：
5.4
通讯作者：
Yu, Kai
中科院分区：
文献类型：
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作者：
Lin, Yilong;Xiong, Wanna;Yu, Kai
民族药理学相关性：中国龙血（CDB），一种从龙血树（Lour.）中提取的药材。陈世诚，历史上已应用于治疗各种疾病，包括溃疡性结肠炎（UC）。不幸的是，潜在的分子机制仍然不清楚。材料与方法：研究CDB对急性UC小鼠模型的影响及结肠组织蛋白质组学变化。以2.5% （wt/vol）葡聚糖硫酸钠（DSS）灌胃Balb/c小鼠，连续8 d建立急性UC模型。UC小鼠灌胃CDB，腹腔注射雷帕霉素（RAPA， mTORC1的特异性抑制剂）后，记录疾病活动指数（DAI）和组织病理学评分。采用等压标签相对和绝对定量（iTRAQ）为基础的LC-MS/MS蛋白质组学技术鉴定结肠组织中的差异表达蛋白（DEPs）。生物信息学分析揭示了DEPs的分子功能和途径。最后采用Western blot和免疫组化方法验证蛋白表达。结果：CDB治疗可显著改善急性UC患者的症状和肠道损伤，而RAPA治疗可导致严重的症状和肠道损伤。对照检查（CK）组和CDB组共有489例dep发生逆转。大多数DEPs富含核糖体结构成分和核糖体途径。CDB处理显著上调mTOR、p-mTOR和p70S6K蛋白的表达，下调Akt、p-Akt和p4EBP1蛋白的表达。然而，与CDB不同，RAPA治疗并没有使mTOR、Akt及其磷酸化形式的水平恢复到接近正常的水平。结论：综上所述，mTOR/核糖体通路功能障碍导致核糖体合成受到抑制在小鼠急性UC的发生发展中发挥了重要作用，CDB而非RAPA通过增强mTOR/核糖体通路的核糖体合成，进而促进蛋白质合成，是治疗急性UC的替代药物。
Ethnopharmacological relevance: Chinese dragon's blood (CDB), a crude drug extracted from Dracaena cochinchinensis (Lour.) S.C. Chen, has been historically applied for the treatment of various diseases, including ulcerative colitis (UC). Unfortunately, the underlying molecular mechanism remains unclear.Materials and methods: In this paper, the effects of CDB treatment on a mouse model of acute UC and proteomic variation in colonic tissue were investigated. The acute UC model in Balb/c mice was induced by administration of 2.5% (wt/vol) dextran sulfate sodium (DSS) in drinking water for 8 days. After the mice with UC were intragastrically administered CDB and intraperitoneally injected with rapamycin (RAPA, a specific inhibitor of mTORC1), the disease activity index (DAI) and histopathological score were recorded. An isobaric tags for relative and absolute quantification (iTRAQ) based LC-MS/MS proteomic technique was adopted to identify the differentially expressed proteins (DEPs) in colonic tissue. Bioinformatics analysis was used to discover the molecular functions and pathways of the DEPs. Finally, Western blot analysis and immunohistochemistry were used to verify the protein expression.Results: The results showed that CDB treatment significantly ameliorated the symptoms and intestinal damage in acute UC, while RAPA treatment led to severe symptoms and intestinal damage. A total of 489 DEPs were reversed in the control check (CK) group and the CDB group. Most DEPs were enriched in the structural constituents of ribosomes and the ribosome pathway. CDB treatment significantly upregulated the expression of the mTOR, p-mTOR and p70S6K proteins and downregulated the expression of the Akt, p-Akt, and p4EBP1 proteins. However, RAPA treatment, unlike CDB, did not return the levels of mTOR, Akt, and their phosphorylated forms to nearly normal.Conclusions: In conclusion, the dysfunction of the mTOR/ribosome pathway resulting in the inhibition of ribosome synthesis played an important role in the development of acute UC in mice, and CDB, but not RAPA, was an alternative drug for the treatment of acute UC by enhancing ribosome synthesis via the mTOR/ribosome pathway and further promoting protein synthesis.