Experimental study on the inhibition of cholesterol gallstone formation by HDCA, the active component of Pulvis Fellis Suis, through regulation of intestinal bile acid metabolism

doi:10.12092/j.issn.1009-2501.2026.08.002

Abstract

Abstract:

AIM: To explore how hyodeoxycholic acid (HDCA) regulates intestinal bile acid metabolism and thereby modulates the activity of farnesoid X receptor (FXR) to inhibit the formation of cholesterol gallstones. METHODS: The C57BL/6J mice were randomly divided into a control group, a cholesterol gallstone model group, and a Pulvis fellis suis (PFS) (7%, 10%) administration intervention group (6 mice in each group). The C57BL/6J mice were also randomly divided into a control group, a cholesterol gallstone model group, and HDCA (0.1%, 0.5%, 1%) administration intervention groups (6 mice in each group). The mice in each group were weighed, photographed, and tissue samples were collected for gallstone grade assessment. The total cholesterol (T-CHO) content in the samples was detected using a total cholesterol detection kit and a microplate reader. In the in vitro cell experiments, the direct regulatory effect of HDCA on FXR in intestinal cells was analyzed by quantitative polymerase chain reaction (qPCR). The differences in fecal microbiota among the groups were detected using 16S ribosomal RNA (16S rRNA) sequencing technology. The changes in the bile acid pool in the ileum tissue were determined by liquid chromatography-mass spectrometry (LC-MS/MS). RESULTS: Compared with the model group, both the 10% PFS and 1% HDCA intervention groups significantly reduced the grade of cholesterol gallstones in the gallbladder and the serum cholesterol content of mice (P<0.01), and the 1% HDCA group significantly decreased the ratio of liver weight to body weight and the cholesterol content in the liver of mice (P<0.01); in the in vitro experiments, HDCA significantly reduced the relative expression level of FXR in intestinal cells Caco-2 (P<0.05, P<0.01); HDCA significantly reduced the content of the Verrucomicrobiota (which was significantly positively correlated with activated FXR-type bile acids) (P<0.05) and significantly increased the content of the Bacteroidota (which was significantly negatively correlated with activated FXR-type bile acids) (P<0.01). Moreover, HDCA significantly changed the composition of intestinal bile acids, reducing the proportion of activated FXR-type bile acids and increasing the proportion of inhibitory FXR-type bile acids (P<0.05). CONCLUSION: HDCA may increase the inhibition of FXR-type bile acids by regulating the intestinal flora, thereby indirectly exerting the effect of inhibiting intestinal FXR, promoting bile acid synthesis and reducing cholesterol saturation, and achieving the goal of inhibiting the formation of cholesterol gallstones.

Key words: hyodeoxycholic acid, Pulvis Fellis Suis, farnesoid X receptor, bile acids, cholesterol gallstones

CLC Number:

R965.2

Yingyang SU, Yan DENG, Xingyu LIU, Yuzhen DU, Min NING, Shan WU, Kaiyue WU, Shuang SHEN. Experimental study on the inhibition of cholesterol gallstone formation by HDCA, the active component of Pulvis Fellis Suis, through regulation of intestinal bile acid metabolism[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2026, 31(8): 1016-1026.

Figures/Tables 8

Fig.1 PFS has a significant preventive effect on cholesterol gallstones in mice ($ \overline{\text{x}} $±s, n=6) A: gallstone grades of the gallbladder in mice; B: incidence of gallstones in mice; C: body weight of mice before model induction; D: body weight of mice at model completion; E: appearance of the gallbladder in mice; F: ratio of liver weight to body weight in mice; G: total cholesterol content in mouse serum. *P<0.05; **P<0.01.

Fig.2 HDCA has a significant preventive effect on cholesterol gallstones in mice ($ \overline{\text{x}} $±s, n=6) A: gallstone grades of the gallbladder in mice; B: incidence of gallstones in mice; C: body weight of mice before model induction; D: body weight of mice at model completion; E: appearance of the gallbladder in mice; F: ratio of liver weight to body weight in mice; G: total cholesterol content in mouse serum; H: total cholesterol content in the liver. *P<0.05; **P<0.01.

Fig.3 HDCA inhibits the expression of FXR in intestinal Caco-2 cells ($ \overline{\text{x}} $±s, n=3). *P<0.05; **P<0.01.

Fig.4 The situation of the intestinal microbiota in mice after HDCA intervention ($ \overline{\text{x}} $±s, n=4) A: heat map showing the relationship between the intestinal microbiota and bile acids; B: composition of the microbial community in the intestinal microbiota; C-K: content of different phyla of microbiota in the intestine. *P<0.05; **P<0.01.

Table 1 The top 10 bile acids in content after HDCA intervention in mice

Bile acid	Content (nmol/g)	Effect on FXR
CA	5931.64	agonists
TCA	2601.21	agonists
HDCA	2598.39	antagonists
DCA	2087.83	agonists
LCA	1473.53	agonists
TDCA	1079.37	agonists
GCA	552.98	agonists
6-ketoLCA	224.95	agonists
beta-MCA	178.26	antagonists
alpha-MCA	120.80	antagonists

Fig.5 The content of different bile acids in mice after HDCA intervention ($ \overline{\text{x}} $±s, n=4) A-G: contents of FXR-activating bile acids; H-J: contents of FXR-inhibiting bile acids. *P<0.05; **P<0.01.

Fig.6 The bile acid conditions of activating or inhibiting FXR after HDCA intervention in mice ($ \overline{\text{x}} $±s, n=4) A: total FXR-activating bile acid content; B: total FXR-inhibiting bile acid content; C: proportion of FXR-activating/inhibiting bile acids. *P<0.05.

Fig.7 The enterohepatic circulation of bile acids

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