Abstract
Indole is produced from dietary tryptophan by tryptophanase in intestinal bacteria, such as Escherichia coli. In the liver, indole is converted into indoxyl sulfate, a uremic toxin and risk factor for chronic kidney disease (CKD). Probiotics and prebiotics are currently used for suppressing CKD, but there are no drugs that directly suppress indole production. In this study, we developed an optimized HPLC method for analyzing indole production and evaluated the effect of diets and rhubarb on indole production via the changes of gut microbiota. In high-carbohydrate and high-fat diet-fed mice, the indole production was significantly higher than in high-fiber diet-fed mice. We further used the high-carbohydrate diet-fed mice as a model for examining the effect of rhubarb on indole production. The 20% methanol-eluted fraction of aqueous rhubarb extract significantly suppressed indole production, and the eluate constituent rhein 8-O-β-d-glucopyranoside (RG) contributed to this effect in a concentration-dependent manner. The effect of RG depended on the anthraquinone core substructure, i.e., the aglycone moiety (rhein) of RG, which appeared to inhibit the tryptophanase function in gut microbiota. Thus, in addition to earlier reports that rhubarb is an effective CKD treatment, our study demonstrated that the anthraquinone moiety in rhubarb prevents uremic toxin production via functional changes in gut microbiota, which suppresses CKD progression.
Graphic abstract
Similar content being viewed by others
References
Japanese Society of Nephrology (2018a) Evidence-based clinical practice guideline for CKD 2018. Japan, Tokyo Igakusha, p 1
Kawakami T, Inagi R, Wada T, Tanaka T, Fujita T, Nangaku M (2010) Indoxyl sulfate inhibits proliferation of human proximal tubular cells via endoplasmic reticulum stress. Am J Physiol Renal Physiol 299:F568–F576. https://doi.org/10.1152/ajprenal.00659.2009
Chiang CK, Tanaka T, Inagi R, Fujita T, Nangaku M (2011) Indoxyl sulfate, a representative uremic toxin, suppresses erythropoietin production in a HIF-dependent manner. Lab Invest 91:1564–1571. https://doi.org/10.1038/labinvest.2011.114
Wood WA, Gunsalus IC, Umbreit WW (1947) Function of pyridoxal phosphate: resolution and purification of the tryptophanase enzyme of Escherichia coli. J Biol Chem 170:313–321
Burns RO, Demoss RD (1962) Properties of tryptophanase from Escherichia coli. Biochim Biophys Acta 65:233–244. https://doi.org/10.1016/0006-3002(62)91042-9
Botsford JL, Demoss RD (1972) Escherichia coli tryptophanase in the enteric environment. J Bacteriol 109:74–80
Banoglu E, Jha GG, King RS (2001) Hepatic microsomal metabolism of indole to indoxyl, a precursor of indoxyl sulfate. Eur J Drug Metab Pharmacokinet 26:235–240. https://doi.org/10.1007/BF03226377
Mishima E, Fukuda S, Mukawa C, Yuri A, Kanemitsu Y, Matsumoto Y, Akiyama Y, Fukuda NN, Tsukamoto H, Asaji K, Shima H, Kikuchi K, Suzuki C, Suzuki T, Tomioka Y, Soga T, Ito S, Abe T (2017) Evaluation of the impact of gut microbiota on uremic solute accumulation by a CE-TOFMS-based metabolomics approach. Kidney Int 92:634–645. https://doi.org/10.1016/j.kint.2017.02.011
Ritz E (2011) Intestinal-renal syndrome: mirage or reality? Blood Purif 31:70–76
Nicholson JK, Holmes E, Kinross J, Burcelin R, Gibson G, Jia W, Pettersson S (2012) Host-gut microbiota metabolic interactions. Science 336:1262–1267. https://doi.org/10.1126/science.1223813
Evenepoel P, Poesen R, Meijers B (2017) The gut-kidney axis. Pediatr Nephrol 32:2005–2014. https://doi.org/10.1007/s00467-016-3527-x
Yoshifuji A, Wakino S, Irie J, Tajima T, Hasegawa K, Kanda T, Tokuyama H, Hayashi K, Itoh H (2016) Gut Lactobacillus protects against the progression of renal damage by modulating the gut environment in rats. Nephrol Dial Transplant 31:401–412. https://doi.org/10.1093/ndt/gfv353
Furuse SU, Ohse T, Jo-Watanabe A, Shigehisa A, Kawakami K, Matsuki T, Chonan O, Nangaku M (2014) Galacto-oligosaccharides attenuate renal injury with microbiota modification. Physiol Rep 2:e12029. https://doi.org/10.14814/phy2.12029
Vaziri ND, Liu SM, Lau WL, Khazaeli M, Nazertehrani S, Farzaneh SH, Kieffer DA, Adams SH, Martin RJ (2014) High amylose resistant starch diet ameliorates oxidative stress, inflammation, and progression of chronic kidney disease. PLoS ONE 9:e114881. https://doi.org/10.1371/journal.pone.0114881
Mitsuma T, Yokozawa T, Oura H, Terasawa K (1987) Rhubarb therapy in patients with chronic renal failure (Part 2). Jpn J Nephrol 29:195–207
Oura H, Zheng PD, Yokozawa T (1984) Effect of onpi-to in rats with chronic renal failure. J Trad Med 1:209–217
Wu XQ, Fujioka K, Yokozawa T, Oura H (1990) Studies on the reciprocal action of component crude drug extract of Ompi-to in rats with renal failure. J Trad Med 7:1–5
Yokozawa T, Suzuki N, Oura H, Nonaka G, Nishioka I (1986) Effect of extracts obtained from rhubarb in rats with chronic renal failure. Chem Pharm Bull 34:4718–4723. https://doi.org/10.1248/cpb.34.4718
Yokozawa T, Chen CP, Tanaka T, Kouno I (1998) Isolation from Wen-Pi-Tang of the active principles possessing antioxidation and radical-scavenging activities. Phytomedicine 5:367–373. https://doi.org/10.1016/S0944-7113(98)80019-6
Zhang Z, Wei F, Vaziri ND, Cheng X, Bai X, Lin R, Zhao Y (2015) Metabolomics insights into chronic kidney disease and modulatory effect of rhubarb against tubulointerstitial fibrosis. Sci Rep 5:14472. https://doi.org/10.1038/srep14472
Takayama K, Matsui E, Kobayashi T, Inoue H, Tsuruta Y, Okamura N (2011) High-performance liquid chromatographic determination and metabolic study of sennoside A in daiokanzoto by mouse intestinal bacteria. Chem Pharm Bull 59:1106–1109. https://doi.org/10.1248/cpb.59.1106
Matsui E, Takayama K, Sato E, Okamura N (2011) The influence of glycyrrhiza and antibiotics on the purgative action of sennoside A from daiokanzoto in mice. Biol Pharm Bull 34:1438–1442. https://doi.org/10.1248/bpb.34.1438
Takayama K, Tsutsumi H, Ishizu T, Okamura N (2012) The influence of rhein 8-O-β-d-glucopyranoside on the purgative action of sennoside A from rhubarb in mice. Biol Pharm Bull 35:2204–2208. https://doi.org/10.1248/bpb.b12-00632
Takayama K, Morita T, Tabuchi N, Fukunaga M, Okamura N (2013) The effect of anthraquinones in daiokanzoto on increasing the synthesis of sennoside A-metabolic enzyme derived from bifidobacteria. J Trad Med 30:215–220. https://doi.org/10.11339/jtm.30.215
Takayama K, Tabuchi N, Fukunaga M, Okamura N (2016) Rhein 8-O-β-d-glucopyranoside elicited the purgative action of daiokanzoto (da-huang-gan-cao-tang), despite dysbiosis by ampicillin. Biol Pharm Bull 39:378–383. https://doi.org/10.1248/bpb.b15-00815
Takayama K, Takahara C, Tabuchi N, Okamura N (2019) Daiokanzoto (Da-Huang-Gan-Cao-Tang) is an effective laxative in gut microbiota associated with constipation. Sci Rep 9:3833. https://doi.org/10.1038/s41598-019-40278-2
Scherzer R, Gdalevsky GY, Goldgur Y, Cohen-Luria R, Bittner S, Parola AH (2009) New tryptophanase inhibitors: towards prevention of bacterial biofilm formation. J Enzyme Inhib Med Chem 24:350–355. https://doi.org/10.1080/14756360802187612
Bolyen E, Rideout JR, Dillon MR, Bokulich NA, Abnet CC, Al-Ghalith GA, Alexander H, Alm EJ, Arumugam M, Asnicar F, Bai Y, Bisanz JE, Bittinger K, Brejnrod A, Brislawn CJ, Brown CT, Callahan BJ, Caraballo-Rodríguez AM, Chase J, Cope EK, Da Silva R, Diener C, Dorrestein PC, Douglas GM, Durall DM, Duvallet C, Edwardson CF, Ernst M, Estaki M, Fouquier J, Gauglitz JM, Gibbons SM, Gibson DL, Gonzalez A, Gorlick K, Guo J, Hillmann B, Holmes S, Holste H, Huttenhower C, Huttley GA, Janssen S, Jarmusch AK, Jiang L, Kaehler BD, Kang KB, Keefe CR, Keim P, Kelley ST, Knights D, Koester I, Kosciolek T, Kreps J, Langille MGI, Lee J, Ley R, Liu YX, Loftfield E, Lozupone C, Maher M, Marotz C, Martin BD, McDonald D, McIver LJ, Melnik AV, Metcalf JL, Morgan SC, Morton JT, Naimey AT, Navas-Molina JA, Nothias LF, Orchanian SB, Pearson T, Peoples SL, Petras D, Preuss ML, Pruesse E, Rasmussen LB, Rivers A, Robeson MS, Rosenthal P, Segata N, Shaffer M, Shiffer A, Sinha R, Song SJ, Spear JR, Swafford AD, Thompson LR, Torres PJ, Trinh P, Tripathi A, Turnbaugh PJ, Ul-Hasan S, van der Hooft JJJ, Vargas F, Vázquez-Baeza Y, Vogtmann E, von Hippel M, Walters W, Wan Y, Wang M, Warren J, Weber KC, Williamson CHD, Willis AD, Xu ZZ, Zaneveld JR, Zhang Y, Zhu Q, Knight R, Caporaso JG (2019) Reproducible, interactive, scalable and extensible microbiome data science using QIIME 2. Nat Biotechnol 37:852–857. https://doi.org/10.1038/s41587-019-0209-9
Martin M (2011) Cutadapt removes adapter sequences from high-throughput sequencing reads. EMBnet J 17:10–12. https://doi.org/10.14806/ej.17.1.200
Amir A, McDonald D, Navas-Molina JA, Kopylova E, Morton JT, Zech XuZ, Kightley EP, Thompson LR, Hyde ER, Gonzalez A, Knight R (2017) Deblur rapidly resolves single-nucleotide community sequence patterns. mSystems 2:e00191-e216. https://doi.org/10.1128/mSystems.00191-16
Quast C, Pruesse E, Yilmaz P, Gerken J, Schweer T, Yarza P, Peplies J, Glöckner FO (2013) The SILVA ribosomal RNA gene database project: improved data processing and web-based tools. Nucl Acids Res 41:D590–D596. https://doi.org/10.1093/nar/gks1219
Carlier JP, Kouas G, Bonne I, Lozniewski A, Mory F (2004) Oribacterium sinus gen nov., sp. nov, within the family ‘Lachnospiraceae’ (phylum Firmicutes). Int J Syst Evol Microbiol 54:1611–1615. https://doi.org/10.1099/ijs.0.63060
Chen YY, Chen DQ, Chen L, Lin JR, Vaziri ND, Guo Y, Zhao YY (2019) Microbiome–metabolome reveals the contribution of gut–kidney axis on kidney disease. J Transl Med 17:5. https://doi.org/10.1186/s12967-018-1756-4
Schulman G, Berl T, Beck GJ, Remuzzi G, Ritz E, Shimizu M, Shobu Y, Kikuchi M (2016) The effects of AST-120 on chronic kidney disease progression in the United States of America: a post hoc subgroup analysis of randomized controlled trials. BMC Nephrol 17:141. https://doi.org/10.1186/s12882-016-0357-9
Cha RH, Kang SW, Park CW, Cha DR, Na KY, Kim SG, Yoon SA, Han SY, Chang JH, Park SK, Lim CS, Kim YS (2016) A randomized, controlled trial of oral intestinal sorbent AST-120 on renal function deterioration in patients with advanced renal dysfunction. Clin J Am Soc Nephrol 11:559–567. https://doi.org/10.2215/CJN.12011214
Sato E, Tanaka A, Oyama J, Yamasaki A, Shimomura M, Hiwatashi A, Ueda Y, Amaha M, Nomura M, Matsumura D, Nakamura T, Node K (2016) Long-term effects of AST-120 on the progression and prognosis of pre-dialysis chronic kidney disease: a 5-year retrospective study. Heart Vessels 31:1625–1632. https://doi.org/10.1007/s00380-015-0785-7
Hatakeyama S, Yamamoto H, Okamoto A, Imanishi K, Tokui N, Okamoto T, Suzuki Y, Sugiyama N, Imai A, Kudo S, Yoneyama T, Hashimoto Y, Koie T, Kaminura N, Saitoh H, Funyu T, Ohyama C (2012) Effect of an oral adsorbent, AST-120, on dialysis initiation and survival in patients with chronic kidney disease. Int J Nephrol 2:376128. https://doi.org/10.1155/2012/376128
Akizawa T, Asano Y, Morita S, Wakita T, Onishi Y, Fukuhara S, Gejyo F, Matsuo S, Yorioka N, Kurokawa K (2009) Effect of a carbonaceous oral adsorbent on the progression of CKD: a multicenter, randomized, controlled trial. Am J Kidney Dis 54:459–467. https://doi.org/10.1053/j.ajkd.2009.05.011
Ueda H, Shibahara N, Takagi S, Inoue T, Katsuoka Y (2007) AST-120, an oral adsorbent, delays the initiation of dialysis in patients with chronic kidney diseases. Ther Apher Dial 11:189–195. https://doi.org/10.1111/j.1744-9987.2007.00430.x
Schulman G, Berl T, Beck GJ, Remuzzi G, Ritz E, Arita K, Kato A, Shimizu M (2015) Randomized placebo-controlled EPPIC Trials of AST-120 in CKD. J Am Soc Nephrol 26:1732–1746. https://doi.org/10.1681/ASN.2014010042
Japanese Society of Nephrology (2018b) Evidence-based clinical practice guideline for CKD 2018. Tokyo Igakusha, Japan, pp 95–96
Wong J, Piceno YM, DeSantis TZ, Pahl M, Andersen GL, Vaziri ND (2014) Expansion of urease- and uricase-containing, indole- and p-cresol-forming and contraction of short-chain fatty acid-producing intestinal microbiota in ESRD. Am J Nephrol 39:230–237. https://doi.org/10.1159/000360010
Hu J, Zhong X, Yan J, Zhou D, Qin D, Xiao X, Zheng Y, Liu Y (2020) High-throughput sequencing analysis of intestinal flora changes in ESRD and CKD patients. BMC Nephrol 21:12. https://doi.org/10.1186/s12882-019-1668-4
Vaziri ND, Wong J, Pahl M, Piceno YM, Yuan J, DeSantis TZ, Ni Z, Nguyen TH, Andersen GL (2013) Chronic kidney disease alters intestinal microbial flora. Kidney Int 83:308–315. https://doi.org/10.1038/ki.2012.345
Roager HM, Licht TR (2018) Microbial tryptophan catabolites in health and disease. Nat Commun 9:3294. https://doi.org/10.1038/s41467-018-05470-4
Turnbaugh PJ, Ley RE, Mahowald MA, Magrini V, Mardis ER, Gordon JI (2006) An obesity-associated gut microbiome with increased capacity for energy harvest. Nature 444:1027–1031. https://doi.org/10.1038/nature05414
Larsen N, Vogensen FK, van den Berg FW, Nielsen DS, Andreasen AS, Pedersen BK, Al-Soud WA, Sørensen SJ, Hansen LH, Jakobsen M (2010) Gut microbiota in human adults with type 2 diabetes differs from non-diabetic adults. PLoS ONE 5:e9085. https://doi.org/10.1371/journal.pone.0009085
Yachida S, Mizutani S, Shiroma H, Shiba S, Nakajima T, Sakamoto T, Watanabe H, Masuda K, Nishimoto Y, Kubo M, Hosoda F, Rokutan H, Matsumoto M, Takamaru H, Yamada M, Matsuda T, Iwasaki M, Yamaji T, Yachida T, Soga T, Kurokawa K, Toyoda A, Ogura Y, Hayashi T, Hatakeyama M, Nakagama H, Saito Y, Fukuda S, Shibata T, Yamada T (2019) Metagenomic and metabolomic analyses reveal distinct stage-specific phenotypes of the gut microbiota in colorectal cancer. Nat Med 25:968–976. https://doi.org/10.1038/s41591-019-0458-7
Poesen R, Windey K, Neven E, Kuypers D, De Preter V, Augustijns P, D’Haese P, Evenepoel P, Verbeke K, Meijers B (2016) The influence of CKD on colonic microbial metabolism. J Am Soc Nephrol 27:1389–1399. https://doi.org/10.1681/ASN.2015030279
Sumida K, Molnar MZ, Potukuchi PK, Thomas F, Lu JL, Matsushita K, Yamagata K, Kalantar-Zadeh K, Kovesdy CP (2017) Constipation and incident CKD. J Am Soc Nephrol 28:1248–1258. https://doi.org/10.1681/ASN.2016060656
Mishima E, Fukuda S, Shima H, Hirayama A, Akiyama Y, Takeuchi Y, Fukuda NN, Suzuki T, Suzuki C, Yuri A, Kikuchi K, Tomioka Y, Ito S, Soga T, Abe T (2015) Alteration of the intestinal environment by lubiprostone is associated with amelioration of adenine-induced CKD. J Am Soc Nephrol 26:1787–1794. https://doi.org/10.1681/ASN.2014060530
Schmidt IM, Hübner S, Nadal J, Titze S, Schmid M, Bärthlein B, Schlieper G, Dienemann T, Schultheiss UT, Meiselbach H, Köttgen A, Flöge J, Busch M, Kreutz R, Kielstein JT, Eckardt KU (2019) Patterns of medication use and the burden of polypharmacy in patients with chronic kidney disease the German Chronic Kidney Disease study. Clin Kidney J 12:663–672. https://doi.org/10.1093/ckj/sfz046
Acknowledgments
The authors are grateful to Mr. Mitsutoshi Mizutani, Ms. Yoko Kada, Mr. Ryoichi Sugihara, Ms. Sayaka Kawamura, and Mr. Yusuke Fukushima for their technical assistance. We would like to thank Editage (www.editage.jp) for English editing.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Takayama, K., Maehara, S., Tabuchi, N. et al. Anthraquinone-containing compound in rhubarb prevents indole production via functional changes in gut microbiota. J Nat Med 75, 116–128 (2021). https://doi.org/10.1007/s11418-020-01459-w
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11418-020-01459-w