背景:高良金,从高良姜(姜科)中提取的生物活性化合物,一种具有重要民族药理学重要性的植物,几千年来一直被用作香料,调味品,和各种条件下的药剂,包括胃肠道疾病.尽管有证据表明其改善胃溃疡的潜力,其抗溃疡特性的分子机制尚不完全清楚。
目的:本研究旨在研究高良姜素对乙醇诱导的小鼠急性胃黏膜损伤(AGMI)的影响,并阐明其分子机制。
方法:将60只BALB/c小鼠随机分为两个主要组:正常对照组(n=10)和乙醇诱导组(n=50)。用40%乙醇和无水乙醇联合建立小鼠AGMI模型后,乙醇诱导组进一步细分为五个亚组(n=10):奥美拉唑对照组(20mg/kg),未经处理的乙醇组,和三个治疗组接受高剂量(50mg/kg)或低剂量(25mg/kg)高良姜素或卡沙平(CPZ,2mg/kg)。通过粘膜损伤指数评价高良姜素的保护作用。苏木精和伊红染色,和炎症标志物的定量(IL-1β,IL-6、IL-8和TNF-α)。使用特定测定试剂盒测量氧化应激水平和基质金属蛋白酶活性。进行分子对接以评估高良姜素与瞬时受体电位香草酸1(TRPV1)途径内关键蛋白的结合亲和力。实时荧光定量PCR(qPCR)检测TRPV1、钙调蛋白(CaM)、P物质(SP),和胃组织中的CGRP。TRPV1、神经生长因子(NGF)的蛋白表达水平,原肌球蛋白受体激酶A(TRKA),转化生长因子β(TGF-β),环氧合酶-2(COX-2),通过蛋白质印迹分析评估核因子κB(NF-κB)。在细胞实验中,在7%乙醇诱导后,用各种浓度的高良姜素处理人胃上皮细胞(GES-1)的培养。细胞增殖,凋亡,使用Hoechst33258染色和transwell迁移测定评估和迁移。用免疫荧光法检测TRPV1蛋白表达,和Bcl-2,BCL2相关X(BAX)的表达水平,和Caspase-3通过qPCR定量。此外,特异性探针试剂盒用于测量细胞内钙离子(Ca2+)和线粒体膜电位。
结果:结果表明,高良姜素通过降低溃疡指数和炎症水平,显著改善了粘膜病理学,同时提高超氧化物歧化酶(SOD)活性和降低丙二醛(MDA)浓度。高良姜素也降低基质金属蛋白酶-2(MMP-2),金属蛋白酶-9(MMP-9)水平,促进粘膜修复。在细胞层面,高良姜素降低细胞内钙离子浓度,减轻线粒体膜电位的下降,增强粘膜细胞的恢复,增加迁移和扩散,减少细胞凋亡。分子上,高良姜素表现出与TRPV1,NGF的有利结合,TRKA,TGF-β,COX-2和NF-κB,并逆转了这些蛋白质的高表达。此外,高良姜素下调TRPV1、CaM、SP,CGRP,巴克斯,和Caspase-3在胃组织/细胞,同时上调Bcl-2mRNA表达。
结论:Galangin通过抑制TRPV1通路的过度激活来缓解AGMI,从而阻断异常的信号转导。这项研究表明,高良姜素对乙醇诱导的AGMI具有治疗潜力,可能是治疗酒精诱导的胃粘膜损伤的可行替代方法。
BACKGROUND: Galangin, a bioactive compound extracted from Alpinia officinarum Hance (Zingiberaceae), a plant with significant ethnopharmacological importance, has been used for thousands of years as a spice, condiment, and medicinal agent for various conditions, including gastrointestinal disorders. Although there is evidence suggesting its potential to improve gastric ulcers, the molecular mechanisms underlying its anti-ulcer properties are not fully understood.
OBJECTIVE: of the Study: This study aimed to investigate the effects of galangin on ethanol-induced acute gastric mucosal injury (AGMI) in mice and elucidate its molecular mechanisms.
METHODS: Sixty BALB/c mice were randomly assigned into two main groups: a normal control group (n = 10) and an ethanol-induced group (n = 50). After establishing the AGMI model in mice using a combination of 40% ethanol and anhydrous ethanol, the ethanol-induced group was further subdivided into five subgroups (n = 10): an omeprazole control group (20 mg/kg), an untreated ethanol group, and three treatment groups receiving high-dose (50 mg/kg) or low-dose (25 mg/kg) galangin or capsazepine (CPZ, 2 mg/kg). The protective effects of galangin were evaluated through mucosal injury indices, hematoxylin and eosin staining, and quantification of inflammatory markers (IL-1β, IL-6, IL-8, and TNF-α). Oxidative stress levels and matrix metalloproteinase activity were measured using specific assay kits. Molecular docking was conducted to assess the binding affinity of galangin to key proteins within the transient receptor potential vanilloid 1 (TRPV1) pathway. Real-time fluorescence quantitative PCR (qPCR) was used to determine mRNA expression levels of TRPV1, calmodulin (CaM), substance P (SP), and CGRP in gastric tissues. Protein expression levels of TRPV1, nerve growth factor (NGF), tropomyosin receptor kinase A (TRKA), transforming growth factor beta (TGF-β), cyclooxygenase-2 (COX-2), and nuclear factor kappa B (NF-κB) were assessed through Western blot analysis. In cellular experiments, Culture of Human Gastric Epithelial Cells (GES-1) were treated with various concentrations of galangin after 7% ethanol induction. Cell proliferation, apoptosis, and migration were evaluated using Hoechst 33258 staining and transwell migration assays. TRPV1 protein expression was detected using immunofluorescence, and the expression levels of Bcl-2, BCL2-Associated X (BAX), and Caspase-3 were quantified by qPCR. Additionally, specific probe kits were used to measure intracellular calcium ions (Ca2+) and mitochondrial membrane potential.
RESULTS: The findings indicate that galangin significantly improved mucosal pathology by reducing ulcer indices and inflammatory levels, while enhancing superoxide dismutase (SOD) activity and decreasing malondialdehyde (MDA) concentration. Galangin also reduced matrix metalloproteinase-2 (MMP-2), m metalloproteinase-9 (MMP-9) levels, promoting mucosal repair. At the cellular level, galangin decreased intracellular calcium ion concentration and mitigated the decline in mitochondrial membrane potential, enhance the restoration of mucosal cells, increased migration and proliferation, and reduced apoptosis. Molecularly, galangin demonstrated favorable binding to TRPV1, NGF, TRKA, TGF-β, COX-2, and NF-κB, and reversed the elevated expression of these proteins. Additionally, galangin downregulated the mRNA expression of TRPV1, CaM, SP, CGRP, BAX, and Caspase-3 in gastric tissues/cells, while upregulating Bcl-2 mRNA expression.
CONCLUSIONS: Galangin mitigates AGMI by inhibiting the overactivation of the TRPV1 pathway, thereby blocking aberrant signal transduction. This study suggests that galangin has therapeutic potential against ethanol-induced AGMI and may be a viable alternative for the treatment of alcohol-induced gastric mucosal injuries.