Wen Yong, Zhan Yu, Li Jun, Xu Ling, Huang Chengzi, Wu Rong, Tang Xuegui
Department of Traditional Chinese Medicine, The Affiliated Hospital of Southwest Medical University Luzhou 646000, Sichuan, China.
Department of Clinical Medicine, Hospital of Chengdu University of Traditional Chinese Medicine Chengdu 610000, Sichuan, China.
Am J Transl Res. 2024 Oct 15;16(10):5829-5845. doi: 10.62347/YLIE1988. eCollection 2024.
This study explores the detailed effects and mechanisms of Zhi Zhu Ma Ren Pill (ZZMRP) on constipation.
Mouse constipation was induced by using loperamide (Lop). The effects and mechanisms of ZZMRP on constipation were addressed by various methods including charcoal meals, hematoxylin and eosin (H&E) staining, immunohistochemistry, enzyme-linked immunosorbent assay (ELISA), terminal deoxynucleotidyl transferase deoxyuridine triphosphate (dUTP) nick end labelling (TUNEL), transmission electron microscopy, and western blot experiments.
Lop-treated mice exhibited delayed transit and reduced ink progradation rates after charcoal meal administration. H&E staining confirmed severe pathologic symptoms in these mice. Additionally, a decline in interstitial Cajal cells (ICCs) was observed in Lop-treated mice, accompanied by reduced concentrations of 5-hydroxytryptamine (5-HT), acetylcholinesterase (AchE), substance P (SP), and vasoactive intestinal peptide (VIP), coupled with an elevated concentration of nitric oxide synthase (NOS). However, ZZMRP treatment markedly ameliorated these changes. In addition, ZZMRP introduction significantly reversed the Lop-induced enhancement in apoptosis rate, endoplasmic reticulum (ER) stress, and levels of proapoptotic proteins and ER stress proteins, and the decrease in the expression of antiapoptotic proteins. These effects were further confirmed with the use of 4-phenylbutyrate (4-PBA), which also reversed the changes in apoptosis rate and protein levels.
ZZMRP alleviates constipation primarily through modulating ER stress-mediated apoptosis in Lop-treated mice, suggesting its use as a therapeutic agent for constipation.
本研究探讨枳术麻仁丸（ZZMRP）对便秘的具体作用及机制。
使用洛哌丁胺（Lop）诱导小鼠便秘。通过多种方法研究ZZMRP对便秘的作用及机制，包括炭末推进实验、苏木精-伊红（H&E）染色、免疫组织化学、酶联免疫吸附测定（ELISA）、末端脱氧核苷酸转移酶介导的缺口末端标记（TUNEL）、透射电子显微镜和蛋白质印迹实验。
给予Lop处理的小鼠在给予炭末后出现转运延迟和墨汁推进率降低。H&E染色证实这些小鼠存在严重的病理症状。此外，在给予Lop处理的小鼠中观察到肠间质 Cajal 细胞（ICC）减少，同时 5-羟色胺（5-HT）、乙酰胆碱酯酶（AchE）、P 物质（SP）和血管活性肠肽（VIP）浓度降低，一氧化氮合酶（NOS）浓度升高。然而，ZZMRP 治疗显著改善了这些变化。此外，引入 ZZMRP 显著逆转了 Lop 诱导的凋亡率升高、内质网（ER）应激以及促凋亡蛋白和 ER 应激蛋白水平的升高，以及抗凋亡蛋白表达的降低。使用 4-苯基丁酸（4-PBA）进一步证实了这些作用，其也逆转了凋亡率和蛋白水平的变化。
ZZMRP 主要通过调节 Lop 处理小鼠中 ER 应激介导的凋亡来缓解便秘，提示其可作为便秘的治疗药物。
