School of Pharmacy, Shanghai University of Medicine and Health Sciences, Shanghai 201318, China.
Yueyang Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai, 200437, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2022 Apr 30;1196:123214. doi: 10.1016/j.jchromb.2022.123214. Epub 2022 Mar 7.
Baicalin (BCL) is a natural compound associated with antioxidant, anti-inflammatory and immunomodulatory activities, among many others. To investigate the therapeutic effect of BCL treatment in ethanol-induced chronic gastritis, we investigated proteomic changes in the gastric tissue to elucidate the therapeutic targets of BCL in chronic gastritis using tandem mass tag (TMT)-based quantitative proteomics.
Sprague-Dawley (SD) rats received 8.7 ml/kg body weight of 56% ethanol intragastrically, which induced chronic gastritis model. Then, BCL (50 mg/kg) or omeprazole (20 mg/kg) was orally administered for 7 days to treat the induced chronic gastritis. After sacrifice, the total protein of the gastric antrum was determined. In addition, a tandem tag labelling for relative and absolute quantification (TMT)-based liquid chromatography with tandem mass spectrometry analysis was performed on the sample to identify differentially expressed proteins (DEPs) between therapy and model groups. Furthermore, the potential protein targets, signalling pathways and protein-protein interaction (PPI) network were analysed using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG). Finally, some potential targets were validated using Western blotting.
A total of 4,452 proteins were identified and quantified in the gastric antrum tissue of SD rats using TMT-based quantitative proteomics. Of these, 107 DEPs, including 44 upregulated and 63 downregulated proteins, were discovered in the BCL-treated group compared with the untreated group with ethanol-induced gastritis, of these were 33 callback proteins. Biological information analysis demonstrated that the selected differential proteins were involved in the enriched pathways, including MAPK, PI3K-Akt and NF-κB. Furthermore, the expression of TPM2, GIMAP4, ICAM-1 and Mpc1 was validated using Western blotting, which was consistent with the proteomics results. The study results confirmed the reliability of the proteomic analysis. Additionally, BCL could decrease the production of interleukin (IL)-2, IL-8 and tumour necrosis factor-α while increasing the expression of epidermal growth factor and B-cell lymphoma-2. Notably, PPI network analysis revealed widespread interactions mediated by BCL.
We investigated the effects and potential mechanism of BCL in chronic gastritis. Proteomic technology was used to explore BCL-affected proteins and some signalling pathways. The results may provide important insights into discovering potential target proteins for treating chronic gastritis.
黄芩苷（BCL）是一种天然化合物，具有抗氧化、抗炎和免疫调节等多种活性。为了研究 BCL 治疗乙醇诱导的慢性胃炎的治疗效果，我们通过串联质量标签（TMT）定量蛋白质组学研究胃组织中的蛋白质组变化，以阐明 BCL 在慢性胃炎中的治疗靶点。
Sprague-Dawley（SD）大鼠给予 56%乙醇 8.7ml/kg 胃内灌胃，诱导慢性胃炎模型。然后，BCL（50mg/kg）或奥美拉唑（20mg/kg）灌胃 7 天治疗诱导的慢性胃炎。处死大鼠后，测定胃窦部总蛋白。此外，对样本进行串联标签标记相对和绝对定量（TMT）-液相色谱-串联质谱分析，以鉴定治疗组和模型组之间的差异表达蛋白（DEPs）。此外，使用基因本体论（GO）和京都基因与基因组百科全书（KEGG）分析潜在的蛋白质靶标、信号通路和蛋白质-蛋白质相互作用（PPI）网络。最后，使用 Western blot 验证一些潜在的靶点。
使用 TMT 定量蛋白质组学在 SD 大鼠胃窦组织中鉴定和定量了 4452 种蛋白质。其中，与未治疗的乙醇诱导性胃炎组相比，BCL 治疗组有 107 个 DEPs，包括 44 个上调和 63 个下调蛋白，其中有 33 个回调蛋白。生物信息学分析表明，所选差异蛋白参与了富集途径，包括 MAPK、PI3K-Akt 和 NF-κB。此外，使用 Western blot 验证了 TPM2、GIMAP4、ICAM-1 和 Mpc1 的表达，结果与蛋白质组学结果一致。研究结果证实了蛋白质组学分析的可靠性。此外，BCL 可降低白细胞介素（IL）-2、IL-8 和肿瘤坏死因子-α的产生，同时增加表皮生长因子和 B 细胞淋巴瘤-2 的表达。值得注意的是，PPI 网络分析揭示了 BCL 介导的广泛相互作用。
我们研究了 BCL 在慢性胃炎中的作用及潜在机制。蛋白质组学技术用于探索 BCL 影响的蛋白质和一些信号通路。研究结果可能为发现治疗慢性胃炎的潜在靶标蛋白提供重要线索。
