Weichang’ an pill (胃肠安丸) alleviates functional dyspepsia through modulating brain-gut peptides and gut microbiota

doi:10.19852/j.cnki.jtcm.2024.06.006

Abstract

Abstract:

OBJECTIVE: To evaluate the effect of Weichang’an pill (胃肠安丸, WCAP) on functional dyspepsia (FD) and explore its regulation of brain-gut peptides (BGPs) and gut microbiota balance as a potential treatment mechanism.

METHODS: The "0 ℃ saline gavage + irregular feeding and tail clamp" method was used to establish the FD rat model, excluding the normal group. The successfully established FD rat models were randomly divided into the model group and the WCAP1 (WC1), WCAP2 (WC2), WCAP3 (WC3), WCAP4 (WC4), WCAP5 (WC5), and Domperidone (Dom) groups (n = 10 per group). The unhandled rats were designated as the control group. The gastrointestinal motility of the rats was evaluated using the charcoal propulsion test. Histopathology was assessed by hematoxylin and eosin (HE) staining. The enzyme-linked immunosorbnent assay method was used to detect the levels of motilin (MTL), gastrin (GAS), vasoactive intestinal peptide (VIP), and somatostatin (SS) in the serum from each group. In addition, the gut microbiota composition of fecal samples was analyzed using 16S rRNA sequencing.

RESULTS: Rat models were successfully established according to data from rat state, gastrointestinal motility assessments, and HE staining. WCAP improved FD symptoms by accelerating the gastric emptying and small intestinal transit of FD rats. Mechanistically, WCAP increased the levels of GAS and MTL and reduced the levels of VIP and SS. Moreover, WCAP treatment restored the total relative abundance of Firmicutes and Bacteroidetes, increased the species richness of the gut flora, and modulated the changes in the composition and function of the gut microbiota.

CONCLUSION: WCAP can effectively promote the recovery of gastrointestinal motility disorders in FD rats. The mechanism may be related to regulating the secretion of BGPs and the composition of the gut microbiota.

Key words: gastrointestinal microbiome, functional dyspepsia, brain-gut peptides, Weichang’an pill

LIAO Mengting, LI Tao, CHU Fuhao, CHEN Yan, LOU Ni, ZHUANG Yuan, BO Rongqiang, DING Xia. Weichang’ an pill (胃肠安丸) alleviates functional dyspepsia through modulating brain-gut peptides and gut microbiota[J]. Journal of Traditional Chinese Medicine, 2024, 44(6): 1177-1186.

Figures/Tables 6

Figure 1 WCAP improves gastrointestinal motility in FD rats A: representative hematoxylin-eosin staining of stomach samples from each group, Magnification × 200, Scar bar = 100 μm. A1: Control group; A2: Model group; A3: WC1 group; A4: WC2 group; A5: WC3 group; A6: WC4 group; A7: WC5 group; A8: Dom group. B: representative appearance of small intestine in each group. B1: Control group; B2: Model group; B3: WC1 group; B4: WC2 group; B5: WC3 group; B6: WC4 group; B7: WC5 group; B8: Dom group. Control group and Model group were given normal saline for seven consecutive days; WC1 group: FD rat treated with 21.60 mg·kg-1·d-1 WCAP for seven days; WC2 group: FD rat treated with 43.20 mg·kg-1·d-1 WCAP for seven days; WC3 group: FD rat treated with 64.80 mg·kg-1·d-1 WCAP for seven days; WC4 group: FD rat treated with 86.40 mg·kg-1·d-1 WCAP for seven days; WC5 group: FD rat treated with 108.00 mg·kg-1·d-1 WCAP for seven days; Dom group: FD rat treated with 2.70 mg·kg-1·d-1 domperidone for seven days. FD: Functional dyspepsia; WCAP: Weichang’an pill; WC1: WCAP1; WC2: WCAP2; WC3: WCAP3; WC4: WCAP4; WC5: WCAP5.

Table 1 Gastric residual rate and Intestinal propulsion rate of rats in each group (%,$\bar{x}±s$)

Group	n	Gastric residual rates	Intestinal propulsion rate
Control	10	54±6	81±5
Model	10	79±6^a	67±5^a
WC1	10	68±12^b	74±5^b
WC2	10	67±4^b	74±5^b
WC3	10	63±14^c	80±5^d
WC4	10	67±12^b	76±8^c
WC5	10	64±13^c	76±9^c
Dom	10	63±11^c	77±11^c

Table 2 Secretion of BGPs in each group (pg/mL, $\bar{x}±s$)

Group	n	MTL	GAS	SS	VIP
Control	10	225±19	160±19	54±5	124±16
Model	10	138±34^a	99±29^a	84±10^a	231±19^a
WC1	10	191±39^b	115±27	80±6	201±24^c
WC2	10	193±31^b	117±32	76±8^d	187±24^b
WC3	10	204±31^b	127±21^c	69±8^b	199±19^b
WC4	10	193±27^b	123±18^d	65±9^b	186±18^b
WC5	10	194±23^b	140±20^b	62±7^b	162±15^b
Dom	10	208±34^b	142±16^b	63±8^b	178±18^b

Figure 2 Effect of WCAP on the diversity of gut microbiota in rats with FD A: venn diagram; B: species accumulation curves; C: Chao diversity index; D: simpson diversity index; E: Shannon diversity index; F: observed-species diversity index. Control: FD rat with normal saline; Model: FD rat with normal saline; WCAP: FD rat treated with 64.80 mg·kg-1·d-1 WCAP for seven days. FD: functional dyspepsia; WCAP: Weichang’an pill; OTU: operational taxonomic units. Dates were analyzed by one-way analysis of variance (n = 10). aP < 0.01, cP < 0.001 versus the control group; bP < 0.01, dP < 0.001, versus the model group.

Figure 3 Effect of WCAP on the structure of the gut microbiota in rats with FD A: phylum level species barplot; B: ratio of Firmicutes/Bacteroidetes; C: class level species barplot; D: order level species barplot; E: genus level species barplot; F: partial least squares discriminant analysis diagram of three groups. Control: FD rat with normal saline; Model: FD rat with normal saline; WCAP: FD rat treated with 64.80 mg·kg-1·d-1 WCAP for seven days. FD: functional dyspepsia; WCAP: Weichang’an pill; PLS-DA: partial least squares discriminant analysis; F/B: Firmicutes/Bacteroidetes.

Figure 4 LEfSe analysis and spearman correlation analysis between gut microbiota and BGPs A: bar graph of LEfSe analysis among the control group, the model group and the WCAP group. B: cladogram graph of LEfSe analysis among the control group, the model group and the WCAP group. C: Spearman correlation analysis was performed between the relative abundance of differential bacteria and the levels of MTL, GAS, SS, VIP. Control: FD rat with normal saline; Model: FD rat with normal saline; WCAP: FD rat treated with 64.80 mg·kg-1·d-1 WCAP for seven days. FD: functional dyspepsia; WCAP: Weichang’an pill; MTL: motilin; GAS: gastrin; SS: somatostatin; VIP: vasoactive intestinal peptide; LEfSe: linear discriminant analysis effect size.

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