摘要
目的基于网络药理学分析和实验探讨龙脷叶水提物治疗便秘的疗效及机制。方法采用在线平台进行网络药理学分析,探讨龙脷叶治疗便秘的分子机制。构建洛哌丁胺诱导的小鼠便秘模型和肠上皮细胞(IEC)损伤模型。采用首次排黑便时间、粪便含水量和胃肠道传输率等指标评估疗效。采用Annexin V-FITC染色评估细胞凋亡情况,JC-1染色检测线粒体膜电位,化学发光法测定三磷酸腺苷(ATP)水平,蛋白质印迹检测相关蛋白表达。结果网络药理学分析显示,龙脷叶中的29种活性成分作用于19个与便秘相关的基因,其中AKT1为关键基因之一。实验结果表明,龙脷叶水提物能有效缓解洛哌丁胺诱导的小鼠便秘症状,包括体质量减轻、肠蠕动减缓、排便时间延长和粪便含水量降低;此外,龙脷叶水提物能抑制IEC-6细胞凋亡,恢复线粒体膜电位,并维持细胞内ATP水平。龙脷叶水提物的治疗机制涉及下调Bcl-2相关X蛋白(Bax)、细胞色素C、裂解的半胱天冬酶-3(Cleaved-Caspase3)和水通道蛋白3(AQP3)的表达,以及增强蛋白激酶B(Akt)磷酸化。结论龙脷叶水提物可有效改善模型小鼠的便秘症状,其机制可能与改善肠道细胞能量代谢、抑制IEC凋亡以及降低AQP3表达有关,提示龙脷叶可作为一种临床治疗便秘的潜在药物。
Objective To explore the efficacy and mechanism of the aqueous extract of Sauropus spatulifolius in treating constipation based on network pharmacology analysis and experiments.Methods Network pharmacology analysis was conducted using an online platform to investigate the molecular mechanism of Sauropus spatulifolius in treating constipation.Loperamide-induced mouse constipation models and intestinal epithelial cell(IEC)injury models were constructed.Therapeutic effects were evaluated using indicators such as the time to the first black stool,fecal water content,and gastrointestinal transit rate.Annexin V-FITC staining was used to assess apoptosis,JC-1 staining was used to detect mitochondrial membrane potential,chemiluminescence was used to measure adenosine triphosphate(ATP)levels,and western blotting was used to detect the expression of relevant proteins.Results Network pharmacology analysis revealed that 29 active components in Sauropus spatulifolius targeted 19 genes associated with constipation,with AKT1 identified as one of the key genes.Experimental results demonstrated that the aqueous extract of Sauropus spatulifolius effectively alleviated loperamide-induced constipation symptoms in mice,including weight loss,reduced intestinal motility,prolonged defecation time,and decreased fecal water content.Additionally,the aqueous extract of Sauropus spatulifolius inhibited IEC-6 cell apoptosis,restored mitochondrial membrane potential,and maintained intracellular ATP levels.The therapeutic mechanism involved downregulating the expression of Bcl-2-associated X protein(Bax),cytochrome C,Cleaved-Caspase3,and aquaporin 3(AQP3),as well as enhancing protein kinase B(Akt)phosphorylation.Conclusion The aqueous extract of Sauropus spatulifolius effectively ameliorates constipation symptoms in mouse models,and its mechanism may be related to improving intestinal cell energy metabolism,inhibiting IEC apoptosis,and reducing AQP3 expression,suggesting that Sauropus spatulifolius could serve as a potential drug for the clinical treatment of constipation.
作者
梁结斐
邱新华
卢俊伟
司文霞
吴伟斌
刘孟华
LIANG Jiefei;QIU Xinhua;LU Junwei;SI Wenxia;WU Weibin;LIU Menghua(School of Pharmacy,Southern Medical University,Guangzhou,Guangdong,510515;School of Pharmacy,Zhaoqing Medical College,Zhaoqing,Guangdong,526020;Department of Neurosurgery,Zhongnan Hospital of Wuhan University,Wuhan,Hubei,430071;Zhaoqing Key Laboratory of Programmed Cell Death and Cancer Research,Zhaoqing Medical College,Zhaoqing,Guangdong,526020)
出处
《实用临床医药杂志》
2025年第17期71-78,共8页
Journal of Clinical Medicine in Practice
基金
广东省中医药局项目(20242130)
广东省普通高校创新团队项目(自然科学)(2024KCXTD058)
广东省普通高校青年创新人才类项目(2023KQNCX268)。
关键词
龙脷叶水提物
便秘
水通道蛋白
蛋白激酶B
肠上皮细胞
线粒体膜电位
网络药理学
洛哌丁胺
aqueous extract of Sauropus spatulifolius
constipation
aquaporin
protein kinase B
intestinal epithelial cell
mitochondrial membrane potential
network pharmacology
loperamide
