Version 1
: Received: 30 December 2023 / Approved: 30 December 2023 / Online: 3 January 2024 (03:37:54 CET)
How to cite:
Li, J.; Fu, Y.; Wang, Y.; Zheng, Y.; Li, Y. scRNA-Seq and Network Pharmacology Analysis Reveal SCF/C-kit as the Targeting Mechanism of Qi Lang Formula Relieving Constipation. Preprints2024, 2024010014. https://doi.org/10.20944/preprints202401.0014.v1
Li, J.; Fu, Y.; Wang, Y.; Zheng, Y.; Li, Y. scRNA-Seq and Network Pharmacology Analysis Reveal SCF/C-kit as the Targeting Mechanism of Qi Lang Formula Relieving Constipation. Preprints 2024, 2024010014. https://doi.org/10.20944/preprints202401.0014.v1
Li, J.; Fu, Y.; Wang, Y.; Zheng, Y.; Li, Y. scRNA-Seq and Network Pharmacology Analysis Reveal SCF/C-kit as the Targeting Mechanism of Qi Lang Formula Relieving Constipation. Preprints2024, 2024010014. https://doi.org/10.20944/preprints202401.0014.v1
APA Style
Li, J., Fu, Y., Wang, Y., Zheng, Y., & Li, Y. (2024). scRNA-Seq and Network Pharmacology Analysis Reveal SCF/C-kit as the Targeting Mechanism of Qi Lang Formula Relieving Constipation. Preprints. https://doi.org/10.20944/preprints202401.0014.v1
Chicago/Turabian Style
Li, J., Yiyuan Zheng and Yong Li. 2024 "scRNA-Seq and Network Pharmacology Analysis Reveal SCF/C-kit as the Targeting Mechanism of Qi Lang Formula Relieving Constipation" Preprints. https://doi.org/10.20944/preprints202401.0014.v1
Abstract
Background: Constipation is one of the chronic gastrointestinal functional diseases that affects the quality of life. While Qi Lang Formula (QLF) has demonstrated effectiveness in alleviating constipation symptoms, its precise mechanism remains elusive.
Methods: QLF was analyzed using UPLC-MS/MS. Targets for QLF were collected from SwissADME, Herb, ITCM databases, and FC-related targets from scRNA-seq and Genecards databases. Overlapping targets suggested potential QLF therapy targets for FC. Enrichment analysis used the KOBAS database. A "drug-ingredient-target" network was constructed with Cytoscape, and AutoDock verified active component binding. H&E staining assessed colonic mucosa changes, TEM examined ICC structural changes. ELISA measured neurotransmitter levels, and western blot verified QLF's effect on target proteins. ICC proliferation was observed through immunofluorescence.
Results: We identified 89 targets of QLF associated with ICC-related FC, with c-Kit emerging as the pivotal target. Molecular docking studies revealed that Atractylenolide Ⅲ, Apigenin, Formononetin, Isorhamnetin, Naringenin, and Ononin exhibited strong binding affinities for the c-Kit structural domain. QLF significantly enhanced first stool passage time, fecal frequency, fecal moisture content, and intestinal propulsion rate. Further analysis unveiled that QLF not only restored neurotransmitter levels but also mitigated colon muscular fiber ruptures. ICC ultrastructure exhibited partial recovery, while RT-qPCR and western blot confirmed upregulated c-Kit expression and downstream targets. Immunofluorescence results indicated ICC proliferation post QLF treatment in rat colon.
Conclusion: Our findings suggest that QLF may promote ICC proliferation by targeting SCF/c-Kit and its downstream signaling pathway, thereby regulating intestinal motility.
Keywords
Qi Lang Formula; Constipation; Interstitial cells of Cajal; SCF/c-Kit; proliferation
Subject
Medicine and Pharmacology, Gastroenterology and Hepatology
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.