Therapeutic role of Wuda granule in gastrointestinal motility disorder through promoting gastrointestinal motility and decreasing inflammatory level

Abstract

Introduction: Previous studies indicated that Wuda Granule (WDG) has been applied in the treatment of gastrointestinal motility disorder (GMD), but the effect and underlying mechanisms is yet to be elucidated. This study aimed to explore the mechanism and pharmacological effect of WDG for GMD via network analysis, verification of animal experiments and clinical experiments. Methods: The chemical components of WDG were identified from the Traditional Chinese Medicine Systems Pharmacology Database (TCMSP, http://lsp.nwu.edu.cn/index.php), and the Encyclopedia of Traditional Chinese Medicine (ETCM, http://www.tcmip.cn/ETCM/index.php/Home/Index/) according to oral bioavailability (OB) ≥ 20% and drug-likeness (DL) ≥ 0.10. The targets of WDG compounds were retrieved from the Swiss Target Prediction database (http://www.swisstargetprediction.ch/) and targets related to GMD were retrieved from GeneCards database (https://www.genecards.org/). Network analysis were performed to screen the key active compounds of WDG and its hub targets. Then the pharmacological effect of WDG were verified via vivo experiments in rats and clinical experiments. Results: The results showed that 117 effective active compounds of WDG were screened and 494 targets of WDG compounds targeting GMD were selected. These targets were involved in the biological process of inflammatory regulation and the regulation of gastrointestinal motility. The mechanism was mainly involved in the regulation of PI3K-Akt signaling pathway and Rap1 signaling pathway. In addition, molecular docking analysis suggested that eight key active compounds of WDG may be mainly responsible for the effect of WDG on GMD by targeting HARS, AKT, and PIK3CA, respectively. Animal experiments and clinical trials both suggested that WDG could exert therapeutical effect on GMD via inhibiting inflammation and promoting gastrointestinal motility, it could also improve digestive function of patients with laparoscopic colorectal cancer after surgery. Conclusion: This study was the first to demonstrate that WDG improved GMD mainly via inhibiting inflammatory level and promoting gastrointestinal motility, providing new insights for the understanding of WDG for GMD, inspiration for future research and reference for clinical strategy in terms of the treatment of GMD.

Keywords: Chinese herb medicine; Wuda granule; gastrointestinal motility disorders; molecule docking; network analysis.

FIGURE 1

WDG may exert therapeutical effect on GMD via multi-compounds and multi-targets. ((A) Herbs-Compounds-Targets network of WDG; (B) Screened active compounds of WDG; (C) Intersection targets between proteins of GMD and targets of WDG compounds; (D) PPI networks of intersection targets; Green: targets related to gastrointestinal motility; Red: targets related to regulation of gastrointestinal smooth muscle contraction; Purple: targets related to regulation of inflammation; (E) 10 hub targets with node degrees 5.64 fold greater than the average node degree; (F) Network of targets related to regulation of inflammation; (G) Network of targets related to regulation of gastrointestinal smooth muscle contraction; (H) Network of targets related to regulation of gastrointestinal motility).

FIGURE 2

WDG may exert therapeutic effect on GMD via the regulation of inflammation and gastrointestinal motility in the GO enrichment of Biological processes (BP), Cellular component (CC), Molecular function (MF). ((A) Regulation of inflammation of biological processes (BP); (B) Regulation of inflammation of biological processes (BP); (C) Results of cellular component (CC); (D) Results of molecular function (MF)).

FIGURE 3

WDG may exert potential therapeutical effect on GMD via regulating multi-pathways. ((A) Results of KEGG pathway enrichment; (B) gene-concept network analysis on KEGG enrichment; and (C) “GMD-pathway” network of pathway crosstalk between PI3K-Akt signaling pathway and Rap1 signaling pathway).

FIGURE 4

WDG decreased inflammation expression and improved gastrointestinal motility in Rats. ((A) The expression level of IL-1β; (B) The expression level of IL-10; (C) The expression level of CRP; (D) The expression level of IL-6; (E) The expression level of TNF $\alpha$ ; (F) The expression level of MDA; (G): The expression level of SOD; (H) The expression level of AGEs; (I) The expression level of VIP; (J) The expression level of Ghrelin; ***: p < 0.001, **: p < 0.01, *: p < 0.05).

FIGURE 5

WDG improved the contraction amplitude, tension and frequency of gastric antrum, duodenum and jejunum in POI rats. ((A) The contraction amplitude of gastric antrum; (B) The contraction tension of gastric antrum; (C) The contraction frequency of gastric antrum; (D) The contraction amplitude of duodenum; (E) The contraction tension of duodenum; (F) The contraction frequency of duodenum; (G) The contraction amplitude of jejunum; (H) The contraction tension of jejunum; (I) The contraction frequency of jejunum; ***: Compared with 1 day after surgery; ###: Compared with before surgery; $∆∆∆$ : Comparison between WDG group and placebo group).

FIGURE 6

WDG improved gastrointestinal motility in patients. ((A) The numbers of MMC in the two groups; (B) The time of MMC phase I; (C) The time of MMC phase II; (D) The contraction amplitude of gastric antrum in MMC phase II (mmHg); (E) The contraction amplitude of duodenum in MMC phase II (mmHg); (F) The contraction amplitude of jejunum in MMC phase II (mmHg); (G) The contractive motility index of gastric antrum in MMC phase II (mmHg/120min); (H) The contractive motility index of duodenum in MMC phase II (mmHg/120min); (I) The contractive motility index of jejunum in MMC phase II (mmHg/120min); ***: Compared with 1 day after surgery; ###: Compared with before surgery; $∆∆∆$ : Comparison between WDG group and placebo group).

FIGURE 7

WDG improved gastrointestinal function and reduced inflammation in patients. ((A) The time of intestinal exhaust (min) in the two groups; (B) The time of intestinal defecation time (min) in the two groups; (C) The duration of postoperative liquid diet (h) after surgery in the two groups; (D) The duration of postoperative semiliquid diet (h) after surgery in the two groups; (E) The expression level of Ghrelin (pg/mL); (F) The expression level of Motilin (pg/mL); (G) The expression level of IL4 (pg/mL); (H) The expression of IL-6 (pg/mL); (I) The expression level of TNF-α (pg/mL); ***: Compared with 1 day after surgery; ###: Compared with before surgery; $∆∆∆$ : Comparison between WDG group and placebo group).

FIGURE 8

The active compounds of WDG had preferable docking ability with Hub-Target of GMD. (A) 3D docking structure of Lauric acid docked with HRAS; (B) 2D docking structure of Lauric acid docked with HRAS; (C) 3D docking structure of 4-Tetradecenoic acid docked with HRAS; (D) 2D docking structure of 4-Tetradecenoic acid docked with HRAS; (E) 3D docking structure of 5Z-tetradecenoic acid docked with HRAS; (F) 2D docking structure of 5Z-tetradecenoic acid docked with HRAS; (G) 3D docking structure of 11-Dodecenoic acid docked with HRAS; (H) 2D docking structure of 11-Dodecenoic acid docked with HRAS; (I) 3D docking structure of beta-sitosterol docked with PIK3CA; (J) 2D docking structure of beta-sitosterol docked with PIK3CA; (K) 3D docking structure of Linoleic acid docked with HRAS; (L) 2D docking structure of Linoleic acid docked with HRAS; (M) Type of interaction between identified compounds and their docking targets; (N) 3D docking structure of Myristelaidic acid docked with HRAS; (O) 2D docking structure of Myristelaidic acid docked with HRAS; (P) 3D docking structure of Stearic acid docked with AKT; (Q) 2D docking structure of Stearic acid docked with AKT; (R) Heat map of docking score of key WDG components docked with Hub-Target of GMD).