| 摘要: |
| 目的 探究干姜黄芩黄连人参汤（GD）对溃疡性结肠炎（UC）的影响及其具体分子机制。方法 使用DSS诱导小鼠UC模型，给药组小鼠分别灌胃给予GD（8g/kg或16g/kg）。记录各组小鼠疾病活动评分（DAI）、口服葡萄糖耐量（OTGG）和结肠长度。使用苏木素伊红（HE）染色观察各组小鼠结肠组织形态。使用LPS构建结肠上皮细胞UC模型。细胞计数试剂盒（CCK-8）用于检测各组细胞活性。流式细胞术用于检测各组细胞凋亡水平。蛋白免疫印迹（WB）检测各组细胞中EGFR和糖酵解相关蛋白表达情况。免疫荧光染色检测各组细胞中EGFR核易位情况。结果 与Control组小鼠相比，DSS组小鼠DAI评分显著升高，OTGG显著升高，结肠长度明显缩短，结肠组织出现明显损伤；与DSS组小鼠相比，GD给药组小鼠DAI评分下降，OTGG下降，结肠长度增长，结肠组织损伤情况改善。体外细胞实验中，与PBS组相比，LPS组细胞活性下降，凋亡水平上升，糖酵解相关蛋白表达增加，p-EGFR表达增加，且在细胞核中表达明显增强；与LPS+NS组相比，LPS+DS组细胞活性上升，凋亡水平下降，糖酵解相关蛋白表达降低，p-EGFR表达下降，且在细胞核中表达减弱。同时，糖酵解抑制剂2-DG的使用加强了GD含药血清对结肠上皮细胞的影响；过表达EGFR增加了细胞中p-EGFR的表达量，逆转了GD含药血清对结肠上皮细胞的影响。结论 GD对UC有明显的治疗效果，且GD可能通过调节EGFR核易位影响葡萄糖代谢，最终缓解UC。 |
| 关键词: 干姜黄芩黄连人参汤 溃疡性结肠炎 葡萄糖代谢 EGFR |
| DOI： |
| 分类号: |
| 基金项目:浙江省中医药科技计划项目（2024ZL078） |
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| Study on the mechanism of regulating EGFR nuclear translocation by Ganjiang Huangqin Huanglian ginseng Decoction affecting glucose metabolism and alleviating ulcerative colitis |
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徐步海, Liu Jiali, Jin Guangjun, Xu Lei
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The Second Affiliated Hospital of Zhejiang Chinese Medical University
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| Abstract: |
| Objective To explore the effect of Ganjiang-Huangqin-Huanglian-renshen Decoction (GD) on ulcerative colitis (UC) and its molecular mechanism.
Methods DSS was used to induce the mouse UC model, and mice in the administration group were given GD (8g/kg or 16g/kg) by intragastric administration. DAI score, oral glucose tolerance (OTGG) and colon length were recorded. The colonic morphology was observed by hematoxylin eosin (HE) staining. Colon epithelial cell UC model was constructed using LPS. Cell Counting Kit (CCK-8) was used to measure cell activity in each group. Flow cytometry was used to detect apoptosis in each group. The expression of related proteins in colon tissues or cells of each group was detected by WB.The nuclear translocation of EGFR in different cell groups was detected by immunofluorescence staining. Results Compared with the Control group, DAI score, OTGG, colon length and colon tissue in DSS group were significantly increased. Compared with mice in DSS group, GD group mice showed decreased DAI scores, reduced OTGG, increased colon length, and improved colon tissue damage. In vitro cell experiments, compared with PBS group, the activity of cells in LPS group decreased, the apoptosis level increased, the expression of glycolytic related protein increased, and the expression of p-EGFR was increased, with a notable enhancement observed in the nucleus. Compared with LPS+NS group, the activity of cells in LPS+DS group increased, the apoptosis level decreased, the expression of glycolytic related proteins decreased, and the expression of p-EGFR was decreased, with a significant reduction observed in the nucleus. At the same time, the use of glycolysis inhibitor 2-DG enhanced the effect of GD drug-containing serum on colon epithelial cells. Overexpression of EGFR increased the expression of p-EGFR in cells and reversed the effect of GD drug-containing serum on colon epithelial cells. Conclusion GD has obvious therapeutic effect on UC. Moreover, GD may affect glucose metabolism by regulating EGFR nuclear translocation, and finally alleviate UC. |
| Key words: Ganjiang-huangqin-huanglian-renshen Decoction Ulcerative colitis Glucose metabolism EGFR |