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艳山姜总黄酮改善无水乙醇致胃黏膜细胞损伤的作用及机制
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| 篇名： | 艳山姜总黄酮改善无水乙醇致胃黏膜细胞损伤的作用及机制 |
| TITLE： | Improvement effect and mechanism of total flavonoids from Alpinia zerumbet on gastric mucosa injury induced by absolute ethanol |
| 摘要： | 目的 探讨艳山姜总黄酮通过微RNA-146a-5p（miR-146a-5p）对无水乙醇诱导的胃黏膜细胞损伤的改善作用及潜在机制。方法以人胃黏膜细胞GES-1为对象，以无水乙醇诱导建立急性胃溃疡细胞模型，在考察不同质量浓度艳山姜总黄酮对细胞活力影响及筛选作用浓度的基础上，检测细胞中miR-146a-5p的相对表达量，细胞中肿瘤坏死因子受体关联因子6（TRAF6）、核因子κBp65（NF-κBp65）、肿瘤坏死因子α（TNF-α）蛋白的表达水平，以及细胞上清液中白细胞介素1β（IL-1β）、IL-6、前列腺素E2（PEG2）水平；验证miR-146a-5p与TRAF6的靶向关系，并观察过表达miR-146a-5p、敲减TRAF6对模型细胞上清液中IL-1β、IL-6、PEG2水平的影响，以及敲减miR-146a-5p对艳山姜总黄酮抗胃溃疡作用的影响。结果与空白组比较，模型组细胞中miR-146a-5p的相对表达量和上清液中PGE2水平均显著降低（P＜0.01），细胞中TRAF6、NF-κBp65、TNF-α蛋白的表达水平和上清液中IL-1β、IL-6水平均显著升高（P＜0.01）。与模型组比较，模型+艳山姜总黄酮（60mg/L）组细胞中miR-146a-5p的相对表达量和上清液中PGE2水平均显著升高（P＜0.01），细胞中TRAF6、NF-κBp65、TNF-α蛋白的表达水平和上清液中IL-1β、IL-6水平均显著降低（P＜0.05或P＜0.01）。miR-146a-5p与TRAF6存在靶向关系且呈负相关；过表达miR-146a-5p或敲减TRAF6后，细胞上清液中IL-1β、IL-6水平均显著降低，PGE2水平显著升高（P＜0.05）。敲减miR-146a-5p后，艳山姜总黄酮作用下的细胞上清液中IL-1β、IL-6水平和细胞中TRAF6蛋白的表达水平均显著升高，PGE2水平显著降低（P＜0.05）。结论艳山姜总黄酮对无水乙醇致胃黏膜细胞损伤有一定的改善作用，其机制可能与上调miR-146a-5p的表达、抑制TRAF6的表达，进而抑制相关炎症因子的分泌有关。 |
| ABSTRACT： | OBJECTIVE To explore the improvement effect and potential mechanism of total flavonoids from Alpinia zerumbet on gastric mucosa injury induced by absolute ethanol through microRNA-146a-5p （miR-146a-5p）. METHODS Using human gastric mucosa GES-1 cells as objects， the acute gastric ulcer model was established by absolute ethanol； based on the investigation of the effects of different concentrations of total flavonoids from A. zerumbet on cell activity and the selection of action concentration， the relative expression level of miR-146a-5p in GES-1 cells was detected， the protein expressions of tumor necrosis factor （TNF） receptor-associated factor 6（TRAF6）， nuclear factor-κB p65 （NF-κB p65） and TNF-α were detected， and the levels of interleukin- 1β （IL-1β）， IL-6 and prostaglandin E2 （PGE2） in cell supernatant were determined. The targeting relationship between miR-146a- 5p and TRAF6 was verified； the effects of overexpressed miR-146a-5p and TRAF6 knockdown on the levels of IL-1β， IL-6 and PEG2 in supernatant of model cells as well as the effects of miR-146a-5p knockdown on anti-gastric ulcer effect of total flavonoids from A. zerumbet were observed. RESULTS Compared with the blank group， the relative expression of miR-146a-5p in cells and the level of PGE2 in cell supernatant were decreased significantly in the model group （P＜0.01）， while the protein expressions of TRAF6， NF-κB p65 and TNF-α in cells and the levels of IL-1β and IL-6 in cell supernatant were increased significantly （P＜ 0.01）. Compared with the model group， the relative expression of miR-146a-5p in cells and the level of PGE2 in cell supernatant were increased significantly in model+A. zerumbet total flavonoids （60 mg/L） group （P＜0.01）， while the protein expressions of TRAF6， NF-κB p65 and TNF-α in cells and 82260767） the levels of IL-1β and IL-6 in cell supernatant were decreased significantly （P＜0.05 or P＜0.01）. There was a targeted relationship and a negative correlation between miR-146a-5p E-mail：3113836821@qq.com and TRAF6. After overexpression of miR-146a-5p or TRAF6 knockdown， the levels of IL-1β and IL-6 were decreased significantly in cell supernatant， while the level of PGE2 was increased significantly （P＜0.05）. After miR-146a-5p knockdown， the levels of IL-1β and IL-6 in cell supernatant and the protein expression of TRAF6 in cells administered with total flavonoids of A. zerumbet were increased significantly， while the level of PGE2 was decreased significantly （P＜0.05）. CONCLUSIONS Total flavonoids of A. zerumbet can improve the gastric mucosa injury induced by absolute ethanol. The mechanism may be related to up-regulating the expression of miR-146a-5p， inhibiting the expression of TRAF6， and further inhibiting the secretion of related inflammatory factors. |
| 期刊： | 2023年第34卷第22期 |
| 作者： | 魏晴；薛娟；梁珊珊 |
| AUTHORS： | WEI Qing，XUE Juan，LIANG Shanshan |
| 关键字： | 艳山姜总黄酮；胃黏膜细胞；微RNA-146a-5p；肿瘤坏死因子受体关联因子6；抗胃溃疡作用 |
| KEYWORDS： | total flavonoids of Alpinia zerumbet； gastric mucosa cell； microRNA-146a-5p； TNF receptor-associated factor 6； |
| 阅读数： | 71 次 |
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