Modified Chaihu Shugan Powder alleviates cholesterol gallstones in mice with liver depression syndrome by regulating gut microbiota and bile acid metabolism

doi:10.12122/j.issn.1673-4254.2026.03.15

Abstract

Abstract:

Objective To investigate the therapeutic effect of modified Chaihu Shugan San (CHSGS) in mice with cholesterol gallstones (CS) and liver depression syndrome and its potential mechanism. Methods Sixty male C57BL/6 mice were randomly divided into blank group (n=9) and model group (n=51) with CS induced by high-lithogenic diet feeding. After verification of successful modeling in 3 mice, the remaining 48 mouse models were divided into CS group, CS-liver depression (CS-LD) group, CHSGS group, and ursodeoxycholic acid (UDCA) group, and the latter 3 groups were subjected to CS-LD modeling by chronic unpredictable mild stress (CUMS) and solitary raising and treated with saline, CHSGS or UDCA gavage for 3 weeks. The mice were assessed for general state, body weight, depressive behavior, ileal pathologies, biliary total cholesterol (TC) and total bile acid (TBA) levels, and ileal mRNA expressions of TGR5, GLP-1, and GLP-2 using q-PCR. 16S rDNA sequencing and metabolomics studies were used to analyze fecal microbiota and bile acids of the mice. Results Compared to the blank control and CS mice, the CS-LD mouse models showed decreased body weight, depressive behaviors, enlarged gallbladder with crystals, intestinal villi damage, increased biliary TC, decreased TBA, and increased ileal GLP-1/2 and TGR5 mRNA expressions, which were obviously improved by CHSGS treatment. Microbiota analysis revealed decreased alpha diversity in CS and CS-LD groups, with altered abundances of Verrucomicrobiota, Akkermansia, Bacteroidota, and Muribaculaceae. Bile acid profiling revealed significant upregulation of TDCA, DCA, and GDCA in CS group and increased CA and 3-ketodeoxycholic acid levels in CS-LD group. Correlation analysis suggested positive correlations between pro-lithogenic bile acids and the harmful gut bacteria. Conclusion Modified CHSGS alleviates liver depression and cholesterol gallstones in mice likely by ameliorating gut microbiota dysbiosis and bile acid metabolism disorders.

Key words: cholesterol gallstone, liver depression syndrome, Chaihu Shugan Powder, intestinal flora

Renjie LIANG, Heying WANG, Zhaoyan LI, Huiping CHEN, Yichun WANG, Li MIN. Modified Chaihu Shugan Powder alleviates cholesterol gallstones in mice with liver depression syndrome by regulating gut microbiota and bile acid metabolism[J]. Journal of Southern Medical University, 2026, 46(3): 615-628.

Figures/Tables 23

References 37

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Primer		Sequence (5'-3')	Length (bp)
GAPDH	F	5'-TGTGTCCGTCGTGGATCTGA-3'	151
GAPDH	R	5'-TTGCTGTTGAAGTCGCAGGAG-3'	151
GLP1/2	F	5'-CCAGAAGAAGTCGCCATTGCC-3'	118
GLP1/2	R	5'-TCAGCCAGTTGATGAAGTCCCT-3'	118
TGR5	F	5'-TTATGGCCTCCTGTTGCCTG-3'	135
TGR5	R	5'-GCCAGGGTTGAGGGTACATC-3'	135

Primer		Sequence (5'-3')	Length (bp)
GAPDH	F	5'-TGTGTCCGTCGTGGATCTGA-3'	151
GAPDH	R	5'-TTGCTGTTGAAGTCGCAGGAG-3'	151
GLP1/2	F	5'-CCAGAAGAAGTCGCCATTGCC-3'	118
GLP1/2	R	5'-TCAGCCAGTTGATGAAGTCCCT-3'	118
TGR5	F	5'-TTATGGCCTCCTGTTGCCTG-3'	135
TGR5	R	5'-GCCAGGGTTGAGGGTACATC-3'	135

Group	Initial body weight (g)	Final body weight (g)
Control	27.40±0.81	28.20±0.54
CS	30.30±1.24^∆∆	31.70±1.11^∆∆
CSD	30.30±1.04^∆∆	26.78±0.90**
ZY	29.82±0.27^∆	29.02±0.80^{# ▲}
XY	29.80±1.93^∆	26.98±1.57

Group	Initial body weight (g)	Final body weight (g)
Control	27.40±0.81	28.20±0.54
CS	30.30±1.24^∆∆	31.70±1.11^∆∆
CSD	30.30±1.04^∆∆	26.78±0.90**
ZY	29.82±0.27^∆	29.02±0.80^{# ▲}
XY	29.80±1.93^∆	26.98±1.57

Group	Time	Total distance (mm)	Center time (s)
Control	Pre-modeling	16036.73±1577.49	43.17±9.59
Control	Post-modeling	15749.01±2721.57	44.93±6.28
CS	Pre-modeling	15449.44±2032.69	37.62±5.28
CS	Post-modeling	15281.21±2350.29	31.71±6.41^∆∆
CSD	Pre-modeling	14860.68±2342.23	42.02±10.07
CSD	Post-modeling	8713.92±850.85**	18.57±5.04*
ZY	Pre-modeling	15541.03±1598.71	29.08±5.29
ZY	Post-modeling	13396.29±482.19^{# ▲}	29.44±3.16^#
XY	Pre-modeling	15811.60±1340.74	31.37±10.77
XY	Post-modeling	7785.30±2518.09	22.64±5.01