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. 2021 Dec;59(1):1452-1463.
doi: 10.1080/13880209.2021.1991383.

Wei Chang An pill regulates gastrointestinal motility in a bidirectional manner

Sitong Jia  1   2   3 , Lijuan Chai  1   2   4 , Jing Zhang  1   2   3 , Min Zhang  1   2   3 , Lin Li  1   2   3 , Yaxin Qi  1   2   3 , Yafen Pang  1   2   3 , Xi Chen  1   2   3 , Nana Fan  1   2   3 , Lin Wang  5 , Yujing Wang  5 , Jixiang Song  5 , Yingjie Sun  5 , Yi Wang  1   2   3 , Lin Miao  1   2   3 , Han Zhang  1   2   4
Affiliations

Wei Chang An pill regulates gastrointestinal motility in a bidirectional manner

Sitong Jia et al. Pharm Biol. 2021 Dec.

Abstract

Context: Wei Chang An (WCA) is a commercial prescription developed for the coordination of gastrointestinal movement.

Objective: To investigate the role of WCA in the regulation of diarrhoea and constipation in rats.

Material and methods: The diarrhoea and constipation models were prepared by gavage of Folium senna and diphenoxylate hydrochloride. Rats were randomized equally (n = 6) into the normal group given saline daily, the positive group given Pinaverium Bromide (13.5 mg/kg) or Sennoside A (0.1 mg/kg) and three WCA-treated groups (22, 44, and 88 mg/kg) by gavage daily for 7 consecutive days. The effects of WCA were assessed by a series of faecal symptoms and histopathology. Gastrointestinal parameters were determined by ELISA. The effect of WCA on gastrointestinal tissues was evaluated by strip assay. Expression of ROCK-1 and MLCK was measured by RT-PCR and Western blotting.

Results: Data from Bristol stool form scale, diarrhoea index, visceral sensitivity, defaecation time, and intestinal propulsive rate showed that WCA protected rats against diarrhoea and constipation (p < 0.01). The up-regulation of Substance P and 5-hydroxytryptamine in diarrhoea rats and down-regulation of Substance P and vasoactive intestinal polypeptide in constipation rats were inhibited by WCA (p < 0.05). WCA stimulated the gastrointestinal strip contractions but inhibited ACh-induced contractions (p < 0.01). The decreased ROCK-1 and MLCK expression in diarrhoea rats and increased in constipation rats were suppressed by WCA (p < 0.01).

Conclusions: WCA has both antidiarrhea and anti-constipation effects, suggesting its bidirectional role in gastrointestinal modulation, and providing evidence of WCA for irritable bowel syndrome treatment.

Keywords: Diarrhoea; Rho-associated coiled-coil forming protein kinase-1 (ROCK-1); constipation; irritable bowel syndrome (IBS); myosin light chain kinase (MLCK).

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Conflict of interest statement

No potential conflict of interest was reported by the author(s).

Figures

Figure 1.
Figure 1.
Effects of WCA on faecal property and cytokine expressions in Folium senna-induced diarrhoea rats. (A) Bristol stool form scale, diarrhoea index (DI) and Visceral sensitivity scores were calculated. (B) The serum levels of NO, SP, MTL and 5-HT in rats from different experimental groups were analyzed by ELISA. Data are represented as mean ± S.E.M. n = 6; #p < 0.05 vs. NC; ##p < 0.01 vs. NC; *p < 0.05 vs. Diarrhoea group; **p < 0.01 vs. diarrhoea group.
Figure 2.
Figure 2.
Effect of WCA on pathological changes in Folium senna-induced diarrhoea rats.HE staining was performed in colonic tissue from rats. The morphology of colonic structure was captured and labelled for intestinal velvet (blue arrows), Goblet cells (red arrows) and crypt (black arrows). Representative image of diarrhoea group, WCA-L, WCA-M, WCA-H and PBT group were shown. (A–F, n = 6). (H) Villus height (µm); (I) Crypt depth (µm); (G) Goblet cells/100 Absorb calls (n = 10). Scale bars 100 µm.
Figure 3.
Figure 3.
Effects of WCA on faecal properties and cytokine expression in CDT-induced constipation rats. (A) Time of first black defaecation, faecal quantity and weight and intestine propulsion were much recorded and calculated. (B) The serum levels of NO, SP, MTL and VIP in rats were analyzed by ELISA. Data are represented as mean ± S.E.M. n = 6; #p < 0.05 vs. NC; ##p < 0.01 vs. NC; *p < 0.05 vs. Constipation group; **p < 0.01 vs. constipation group.
Figure 4.
Figure 4.
Effect of WCA on pathological changes in CDT-induced constipation rats. HE staining was performed in colonic tissue from rats. The morphology of colonic structure was captured and labelled for intestinal velvet (blue arrows), Goblet cells (red arrows) and crypt (black arrows). Representative image of constipation group, WCA-L, WCA-M, WCA-H and sennoside A group were shown. (A–F, n = 6). (H) villus height (µm); (i) crypt depth (µm); (g) goblet cells/100 absorb calls (n = 10). Scale bars 100 µm.
Figure 5.
Figure 5.
The inhibitory effect of WCA on ACh-induced contractions of rat antrum, jejunum and ileum. (A n = 1) Rat jejunum strips were isolated and stimulated with different concentrations of ACh. (B, C, D) Strips from rat antrum (B), jejunum (C) and ileum (D) were isolated and stimulated with ACh (10 µM). After the stable induction of contraction, WCA was added and the contractile force and frequency of spontaneous contractions were recorded. Data are represented as mean ± S.E.M. n = 6.
Figure 6.
Figure 6.
The effects of WCA on contractions of rat antrum, jejunum and ileum. Strips from rat antrum (A), jejunum (B) and ileum (C) were isolated and stimulated with WCA. The contractile force and frequency of spontaneous contractions were recorded. Data are represented as mean ± S.E.M. n = 6.
Figure 7.
Figure 7.
WCA inhibited up-regulation of ROCK-1 and MLCK expressions in diarrhoea rat. (A) Total RNA was extracted and RT-PCR was performed to detect the relative mRNA expression of ROCK-1. (B) Protein was extracted and Western blotting was performed to detect the protein expressions of ROCK-1 and MLCK in antrum and jejunum. The images shown are representative of three independent experiments. Data are represented as mean ± S.E.M. ##p < 0.01 vs. NC; *p < 0.05 vs. diarrhoea group.
Figure 8.
Figure 8.
WCA enhanced down-regulation of ROCK-1 and MLCK expression in constipation rats. (A) Total RNA was extracted and RT-PCR was performed to detect the relative mRNA expression of ROCK-1. (B) Protein was extracted and Western blotting was performed to detect the protein expression of ROCK-1 and MLCK in antrum and jejunum. The images shown are representative of three independent experiments. Data are represented as mean ± S.E.M. #p < 0.05 vs. NC; *p < 0.05 vs. constipation group; **p < 0.01 vs. constipation group.
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Grants and funding

This research was funded by Tianjin Municipal Education Commission Scientific Research Project [Natural Science, No. 2018ZD01], Science and Technology Key Program of Tianjin [No. 20ZYJDJC00070] and Scientific research project in key fields of traditional Chinese medicine of Tianjin Health Commission [No. 202-003].