Gut Microbiota, a Potential Mediated Target for Reducing Geniposide Hepatotoxicity by Interacting with Isoflavones

Wen Yang; Wen Zhang; Xinhui Huang; Shuwen Geng; Yujia Zhai; Yuetong Jiang; Tian Tian; Yuye Gao; Jing He; Taohong Huang; Yunxia Li; Wenjing Zhang; Jun Wen; Jian-lin Wu; Guangji Wang; Tingting Zhou

doi:10.1016/j.eng.2024.10.023

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Engineering ›› 2025, Vol. 47 ›› Issue (4) : 222-235. DOI: 10.1016/j.eng.2024.10.023

Research

Article

Gut Microbiota, a Potential Mediated Target for Reducing Geniposide Hepatotoxicity by Interacting with Isoflavones

Wen Yang^a^,^b^,^# ,
Wen Zhang^a^,^b^,^# ,
Xinhui Huang^a^,^b^,^# ,
Shuwen Geng^a^,^b ,
Yujia Zhai^c ,
Yuetong Jiang^a^,^b ,
Tian Tian^a^,^b ,
Yuye Gao^a^,^b ,
Jing He^a^,^b ,
Taohong Huang^d ,
Yunxia Li^e ,
Wenjing Zhang^f ,
Jun Wen^a^,^b ,
Jian-lin Wu^g ,
Guangji Wang^h^,^* ,
Tingting Zhou^a^,^b^,^*

Author information +

History +

Highlights

• Clarifying the interplay of gut microbiota, enzyme and geniposide metabolism.

• Unearthing the role of Lactobacillus spp. in alleviating drug-induced liver injury.

• Clinical study on the regulatory role of isoflavone-rich diets on Lactobacillus.

Abstract

Geniposide, the principal active iridoid glucoside ingredient in Fructus gardeniae used in numerous traditional Chinese clinical prescriptions, has been shown to cause herbal hepatotoxicity because of its glycone metabolite genipin. This study explored the role of gut microbiota in alleviating geniposide hepatotoxicity with isoflavones in soy products. Metabolic profiling using ultra high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-Q/TOF-MS) revealed two metabolic pathways and six main forms of geniposides in vivo. Enzyme inhibitor experiments have shown that isoflavones alter geniposide metabolism by mediating specific enzymes, including β-glucosidase (β-GC) and sulfotransferase (SULT), in an established pseudo-sterile rat model. Isoflavones pretreatment by gavage for three weeks optimized the structure of the gut microbiota was linked to the regulation of key metabolic enzymes. Furthermore, experiments involving fecal microbiota transplantation (FMT) established the direct contribution of the gut microbiota to the regulation of enzyme activities and geniposide metabolism. This study demonstrated that isoflavones in soy products regulated the metabolic enzymes of geniposode dependent on gut microbiota, especially Lactobacillus spp., which was further verified in our clinical trials analyzed using 16S ribosomal RNA (rRNA) and metagenomic sequencing, thus regulating geniposide metabolism. Furthermore, as dominant beneficial bacterium, Lactobacillus spp. were discovered to be promising microbial targets for the better management of geniposide hepatotoxicity. These findings provide valuable insights for the prevention and intervention of drug-induced liver injury.

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Keywords

Geniposide / Isoflavones rich diet / Gut microbiota / Fecal microbiota transplantation / Intestinal metabolic enzymes / Drug-induced liver injury

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Wen Yang, Wen Zhang, Xinhui Huang, Shuwen Geng, Yujia Zhai, Yuetong Jiang, Tian Tian, Yuye Gao, Jing He, Taohong Huang, Yunxia Li, Wenjing Zhang, Jun Wen, Jian-lin Wu, Guangji Wang, Tingting Zhou. Gut Microbiota, a Potential Mediated Target for Reducing Geniposide Hepatotoxicity by Interacting with Isoflavones. Engineering, 2025, 47(4): 222‒235 https://doi.org/10.1016/j.eng.2024.10.023

References

Publishing order | Descend order by publishing year | Descend order by cited within

[1]	Guo H, Liu X, Jiang Y, He J, Ge W, Hao H, et al.Characterization and quantification of the Chinese medical formula Zhi-Zi-Chi decoction, a systematic strategy for the attenuation and synergy of compatibility mechanism.J Pharm Biomed Anal 2023; 223:115130.

[2]	Wang L, Hu Z, Yang W, Loo SKF, Ip SP, Xian YF, et al.Anti-atopic dermatitis effect of a modified Huang-Lian-Jie-Du decoction and its active fraction on 2,4-dinitrobenzene and MC903-induced mouse models.Phytomedicine 2022; 104:154346.

[3]	Sung YY, Kim HK.Crocin ameliorates atopic dermatitis symptoms by down regulation of Th2 response via blocking of NF-κB/STAT6 signaling pathways in mice.Nutrients 2018; 10(11):1625.

[4]	Ran D, Hong W, Yan W, Mengdie W.Properties and molecular mechanisms underlying geniposide-mediated therapeutic effects in chronic inflammatory diseases.J Ethnopharmacol 2021; 273:113958.

[5]	Zhou L, Bao L, Wang Y, Chen M, Zhang Y, Geng Z, et al.An integrated analysis reveals geniposide extracted from Gardenia jasminoides J.Ellis regulates calcium signaling pathway essential for influenza a virus replication.Front Pharmacol 2021; 12:755796.

[6]	Moreira J, Machado M, Dias-Teixeira M, Ferraz R, Delerue-Matos C, Grosso C.The neuroprotective effect of traditional Chinese medicinal plants—a critical review.Acta Pharm Sin B 2023; 13(8):3208-3237.

[7]	Ge N, Yan G, Sun H, Yang L, Kong L, Sun Y, et al.Version updates of strategies for drug discovery based on effective constituents of traditional Chinese medicine.Acupunct Herb Med 2023; 3(3):158-179.

[8]	Shan M, Yu S, Yan H, Guo S, Xiao W, Wang Z, et al.A review on the phytochemistry, pharmacology, pharmacokinetics and toxicology of geniposide, a natural product.Molecules 2017; 22(10):1689.

[9]	De G, Chen A, Zhao Q, Xie R, Wang C, Li M, et al.A multi-herb-combined remedy to overcome hyper-inflammatory response by reprogramming transcription factor profile and shaping monocyte subsets.Pharmacol Res 2021; 169:105617.

[10]	Su L, Mao J, Hao M, Lu T, Mao C, Ji D, et al.Integrated plasma and bile metabolomics based on an UHPLC-Q/TOF-MS and network pharmacology approach to explore the potential mechanism of schisandra chinensis-protection from acute alcoholic liver injury.Front Pharmacol 2020; 10:1543.

[11]	Li Y, Pan H, Li X, Jiang N, Huang L, Lu Y, et al.Role of intestinal microbiota-mediated genipin dialdehyde intermediate formation in geniposide-induced hepatotoxicity in rats.Toxicol Appl Pharmacol 2019; 377:114624.

[12]	Luo Y, Zhang X, Zhang W, Yang Q, You W, Wen J, et al.Compatibility with Semen Sojae Praeparatum attenuates hepatotoxicity of Gardeniae Fructus by regulating the microbiota, promoting butyrate production and activating antioxidant response.Phytomedicine 2021; 90:153656.

[13]	Chang R, Liu J, Luo Y, Huang T, Li Q, Wen J, et al.Isoflavones’ effects on pharmacokinetic profiles of main iridoids from Gardeniae Fructus in rats.J Pharm Anal 2020; 10(6):571-580.

[14]	Liu J, Chang R, Zhang X, Wang Z, Wen J, Zhou T.Non-isoflavones diet incurred metabolic modifications induced by constipation in rats via targeting gut microbiota.Front Microbiol 2018; 9:3002.

[15]	Ghimire S, Cady NM, Lehman P, Peterson SR, Shahi SK, Rashid F, et al.Dietary isoflavones alter gut microbiota and lipopolysaccharide biosynthesis to reduce inflammation.Gut Microbes 2022; 14(1):2127446.

[16]	Gong X, Li X, Bo A, Shi R, Li Q, Lei L, et al.The interactions between gut microbiota and bioactive ingredients of traditional Chinese medicines: a review.Pharmacol Res 2020; 157:104824.

[17]	Lu YM, Xie JJ, Peng CG, Wang BH, Wang KC, Li LJ.Enhancing clinical efficacy through the gut microbiota: a new field of traditional Chinese medicine.Engineering 2019; 5(1):40-49.

[18]	Meng N, Lyu Y, Zhang X, Chai X, Li K, Wang Y.The exciting and magical journey of components from compound formulae to where they fight.Acupunct Herb Med 2022; 2(4):240-252.

[19]	Luo Y, Gao F, Chang R, Zhang X, Zhong J, Wen J, et al.Metabolomics based comprehensive investigation of Gardeniae Fructus induced hepatotoxicity.Food Chem Toxicol 2021; 153:112250.

[20]	Wu H, Li X, Yan X, An L, Luo K, Shao M, et al.An untargeted metabolomics-driven approach based on LC-TOF/MS and LC-MS/MS for the screening of xenobiotics and metabolites of Zhi-Zi-Da-Huang decoction in rat plasma.J Pharm Biomed Anal 2015; 115:315-322.

[21]	Jiang P, Ma Y, Gao Y, Li Z, Lian S, Xu Z, et al.Comprehensive evaluation of the metabolism of genipin-1-β-D-gentiobioside in vitro and in vivo by using HPLC-Q-TOF.J Agric Food Chem 2016; 64(27):5490-5498.

[22]	Wang G, Bao B, Han Z, Han Q, Yang X.Metabolic profile of Fructus Gardeniae in human plasma and urine using ultra high-performance liquid chromatography coupled with high-resolution LTQ-orbitrap mass spectrometry.Xenobiotica 2016; 46(10):901-912.

[23]	Yu J, Guo X, Zhang Q, Peng Y, Zheng J.Metabolite profile analysis and pharmacokinetic study of emodin, baicalin and geniposide in rats.Xenobiotica 2018; 48(9):927-937.

[24]

Zhang

, Pi

, Zheng

, Liu

, Song

.Comprehensive investigation of in-vivo ingredients and action mechanism of iridoid extract from Gardeniae Fructus by liquid chromatography combined with mass spectrometry, microdialysis sampling and network pharmacology.J Chromatogr B Analyt Technol Biomed Life Sci 2018; 1076:70-76.

[25]	Zhou J, Zhang Y, Li N, Zhao D, Lu Y, Wang L, et al.A systematic metabolic pathway identification of common gardenia fruit (Gardeniae Fructus) in mouse bile, plasma, urine and feces by HPLC-Q-TOF-MS/MS.J Chromatogr B Analyt Technol Biomed Life Sci 2020; 1145:122100.

[26]

Han

, Yang

, Xu

, Ding

, Annie

Bligh, Zhang

, et al.Identification of metabolites of geniposide in rat urine using ultra-performance liquid chromatography combined with electrospray ionization quadrupole time-of-flight tandem mass spectrometry.Rapid Commun Mass Spectrom 2011; 25(21):3339-3350.

[27]	Gao Y, Fu Y, Li N, Jiang Y, Liu X, Gao C, et al.Carboxyl-containing components delineation via feature-based molecular networking: a key to processing conditions of fermented soybean.Food Chem 2023; 423:136321.

[28]	Wu W, Lv L, Shi D, Ye J, Fang D, Guo F, et al.Protective effect of Akkermansia muciniphila against immune-mediated liver injury in a mouse model.Front Microbiol 2017; 8:1804.

[29]	Liu L, Wu Q, Chen Y, Gu G, Gao R, Peng B, et al.Updated pharmacological effects, molecular mechanisms, and therapeutic potential of natural product Geniposide.Molecules 2022; 27(10):3319.

[30]	Wang Y, Feng F.Evaluation of the hepatotoxicity of the Zhi-Zi-Hou-Po decoction by combining UPLC-Q-Exactive-MS-based metabolomics and HPLC-MS/MS-based geniposide tissue distribution.Molecules 2019; 24(3):511.

[31]	Zhu H, Bi K, Han F, Guan J, Zhang X, Mao X, et al.Identification of the absorbed components and metabolites of Zhi-Zi-Da-Huang decoction in rat plasma by ultra-high performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry.J Pharm Biomed Anal 2015; 111:277-287.

[32]	Akao T, Kobashi K, Aburada M.Enzymic studies on the animal and intestinal bacterial metabolism of geniposide.Biol Pharm Bull 1994; 17(12):1573-1576.

[33]	Zhang L, Li F, Qin WJ, Fu C, Zhang XL.Changes in intestinal microbiota affect metabolism of ginsenoside Re.Biomed Chromatogr 2021; 35(8):e5189.

[34]	Huang Z, Xu Y, Wang Q, Gao X.Metabolism and mutual biotransformations of anthraquinones and anthrones in rhubarb by human intestinal flora using UPLC-Q-TOF/MS.J Chromatogr B Analyt Technol Biomed Life Sci 2019; 1104:59-66.

[35]	Hou YC, Tsai SY, Lai PY, Chen YS, Chao PDL.Metabolism and pharmacokinetics of genipin and geniposide in rats.Food Chem Toxicol 2008; 46(8):2764-2769.

[36]	Pala L, Sirec T, Spitz U.Modified enzyme substrates for the detection of bacteria: a review.Molecules 2020; 25(16):3690.

[37]	Yeo SK, Liong MT.Growth, bioconversion of isoflavones and probiotic properties of parent and subsequent passages of Lactobacillus upon ultraviolet radiation.Int J Food Sci Nutr 2012; 63(7):821-831.

[38]	Sestelo A, Poza M, Villa TG.β-glucosidase activity in a Lactobacillus plantarum wine strain.World J Microb Biot 2004; 20(6):633-637.

[39]	Yang L, Bajinka O, Jarju PO, Tan Y, Taal AM, Ozdemir G.The varying effects of antibiotics on gut microbiota.AMB Express 2021; 11(1):116.

[40]	Zhang M, Wang Y, Wu Y, Li F, Han M, Dai Y, et al.In vitro transformation of protopanaxadiol saponins in human intestinal flora and its effect on intestinal flora.Evid Based Complement Alternat Med 2021; 2021:1735803.

[41]	Bai X, Fu R, Liu Y, Deng J, Fei Q, Duan Z, et al.Ginsenoside Rk3 modulates gut microbiota and regulates immune response of group 3 innate lymphoid cells to against colorectal tumorigenesis.J Pharm Anal 2023; 14(2):259-275.

[42]	Yang Y, Wang Y, Zhao L, Wang F, Li M, Wang Q, et al.Chinese herbal medicines for treating ulcerative colitis via regulating gut microbiota–intestinal immunity axis.Chin Herb Med 2023; 15(2):181-200.

[43]	Zhang K, Chen L, Yang J, Liu J, Li J, Liu Y, et al.Gut microbiota-derived short-chain fatty acids ameliorate methamphetamine-induced depression- and anxiety-like behaviors in a Sigmar-1 receptor-dependent manner.Acta Pharm Sin B 2023; 13(12):4801-4822.

[44]	Beam A, Clinger E, Hao L.Effect of diet and dietary components on the composition of the gut microbiota.Nutrients 2021; 13(8):2795.

[45]	Gentile CL, Weir TL.The gut microbiota at the intersection of diet and human health.Science 2018; 362(6416):776-780.

[46]	Weersma RK, Zhernakova A, Fu J.Interaction between drugs and the gut microbiome.Gut 2020; 69(8):1510-1519.

[47]	Yu H, Xu H, Yang X, Zhang Z, Hu J, Lu J, et al.Gut microbiota-based pharmacokinetic-pharmacodynamic study and molecular mechanism of specnuezhenide in the treatment of colorectal cancer targeting carboxylesterase.J Pharm Anal 2023; 13(9):1024-1040.

[48]	Cao H, Zhu Y, Hu G, Zhang Q, Zheng L.Gut microbiome and metabolites, the future direction of diagnosis and treatment of atherosclerosis?.Pharmacol Res 2023; 187:106586.

[49]	Wang Y, Shou JW, Li XY, Zhao ZX, Fu J, He CY, et al.Berberine-induced bioactive metabolites of the gut microbiota improve energy metabolism.Metabolism 2017; 70:72-84.

[50]	Stasi C, Sadalla S, Milani S.The relationship between the serotonin metabolism, gut–microbiota and the gut–brain axis.Curr Drug Metab 2019; 20(8):646-655.

[51]	Quaranta G, Sanguinetti M, Masucci L.Fecal microbiota transplantation: a potential tool for treatment of human female reproductive tract diseases.Front Immunol 2019; 10:2653.

[52]	Zhao Z, Ning J, Bao X, Shang M, Ma J, Li G, et al.Fecal microbiota transplantation protects rotenone-induced Parkinson’s disease mice via suppressing inflammation mediated by the lipopolysaccharide-TLR4 signaling pathway through the microbiota–gut–brain axis.Microbiome 2021; 9(1):226.

[53]	Baruch EN, Youngster I, Ben-Betzalel G, Ortenberg R, Lahat A, Katz L, et al.Fecal microbiota transplant promotes response in immunotherapy-refractory melanoma patients.Science 2021; 371(6529):602-609.

[54]	Munro IC, Harwood M, Hlywka JJ, Stephen AM, Doull J, Flamm WG, et al.Soy isoflavones: a safety review.Nutr Rev 2003; 61(1):1-33.

[55]	Wang C, Chen J, Tian W, Han Y, Xu X, Ren T, et al.Natto: a medicinal and edible food with health function.Chin Herb Med 2023; 15(3):349-359.

[56]	Bhagwat S, Haytowitz DB, Holden JM.USDA database for the isoflavone content of selected foods [Release 2.0]. Beltsville: US Department of Agriculture; 2008.