<sup>1</sup>H‑NMR‑based metabolic profiling of rat urine to assess the toxicity‑attenuating effect of the sweat‑soaking method on Radix <em>Wikstroemia indica</em>

Zhou,Zhi-Rong; Feng,Guo; Li,Lai-Lai; Li,Wei; Wu,Zhen-Guang; Zheng,Chuan-Qi; Xu,Qin; Ren,Chen-Chen; Peng,Li-Zhen

doi:10.3892/etm.2022.11392

¹H‑NMR‑based metabolic profiling of rat urine to assess the toxicity‑attenuating effect of the sweat‑soaking method on Radix Wikstroemia indica

Authors:
- Zhi-Rong Zhou
- Guo Feng
- Lai-Lai Li
- Wei Li
- Zhen-Guang Wu
- Chuan-Qi Zheng
- Qin Xu
- Chen-Chen Ren
- Li-Zhen Peng
View Affiliations

Affiliations: Department of Chinese Materia Medica, Guizhou University of Traditional Chinese Medicine, Guiyang, Guizhou 550025, P.R. China
Published online on: May 25, 2022 https://doi.org/10.3892/etm.2022.11392
Article Number: 465
Copyright: © Zhou et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Radix Wikstroemia indica (L.) C.A. Mey. (RWI) is a toxic medicinal species primarily present in the Miao area of China. The toxicity of RWI is effectively reduced whilst maintaining the therapeutic effect when processed using the ‘sweat‑soaking method’, which is a common method of Traditional Chinese Medicine preparation. However, there is a lack of scientific and medical evidence to explain the potential mechanisms by which the toxicity of RWI is reduced after preparation using this method, and the endogenous systemic metabolic effect of RWI remains uncertain. The aim of the present study was to explore the endogetnous metabolic alterations caused by RWI and to examine the possibility of reducing the toxicity of RWI using the sweat‑soaking method using proton nuclear magnetic resonance (NMR) metabolomic analysis in rats. Principal Component Analysis, Partial Least Squares‑Discriminant Analysis (PLS‑DA) and Orthogonal PLS‑DA were used to assess individual proton NMR spectra. A total of 34 metabolic products were altered after delivering raw RWI, and 32 endogenous metabolites were induced by processed RWI. The metabolic pathways that lead to a significant impact on energy and carbohydrate, amino acid, organic acids and lipid metabolism following raw and processed RWI use were identified. The mitochondria of hepatic and renal tubules of rats were injured in the raw RWI group, whereas the processed product reduced or interfered with energy substrate, carbohydrate and amino acid metabolism, whilst reducing the levels of metabolic markers of hepatotoxicity and nephrotoxicity, without causing damage to the mitochondria. Our previous study showed that the median lethal dose (LD₅₀) value of raw RWI was 4.05 g/kg in rats after oral administration; however, the LD₅₀ value of the processed RWI could not be measured. The maximum tolerated dose and minimum lethal dose were 20 and 30 g/kg for the processed RWI, respectively, corresponding to 109 and 164 times the clinical daily dose (0.029 g/kg). Thus, the sweat‑soaking method reduced the toxicity of RWI. Moreover, after processing, the toxic component YH‑10 was converted into a YH‑10 + OH compound, reducing the content of the toxic YH‑10 by 48%, whilst also reducing the contents of the toxic components YH‑12 and YH‑15 by 44 and 65%, respectively. In conclusion, the present study showed that the sweat‑soaking method reduced the toxicity of RWI, as evidenced by the reduction of the levels of metabolic markers and the activity of metabolic pathways, thus providing a basis for processing of RWI for clinical use.

View Figures

View References

1	Ho WS, Xue JY, Sun SM, Ooi VE and Li YL: Antiviral activity of daphnoretin isolated from Wikstroemia indica. Phytother Res. 24:657–661. 2010.PubMed/NCBI View Article : Google Scholar
2	Zhang QR and Xia GC: Colored illustrated book of poisonous chinese herbal medicine. Tianjin Science and Technology Translation Publishing Company. 131:2006.
3	Kato M, He YM, Dibwe DF, Li F, Awale S, Kadota S and Tezuka Y: New guaian-type sesquiterpene from Wikstroemia indica. Nat Prod Commun. 9:1–2. 2014.PubMed/NCBI
4	Tong LJ, Sun LX, Sun LX, LU XF and XU CL: Isolation and structural identification of the chemical constituents from the rhizomes of Wikstroemia indica. Chin J Med Chemistry. 25:50–53. 2015.
5	Chang H, Wang YW, Gao X, Song Z, Awale S, Han N, Liu Z and Yin J: Lignans from the root of Wikstroemia indica and their cytotoxic activity against PANC-1 human pancreatic cancer cells. Fitoterapia. 121:31–37. 2017.PubMed/NCBI View Article : Google Scholar
6	Wang GC, Zhang XL, Wang YF, Li GQ, Ye WC and Li YL: Four new dilignans from the roots of Wikstroemia indica. Chem Pharm Bull (Tokyo). 60:920–923. 2012.PubMed/NCBI View Article : Google Scholar
7	Shao M, Huang XJ, Liu JS, Han WL, Cai HB, Tang QF and Fan Q: A new cytotoxic biflavonoid from the rhizome of Wikstroemia indica. Nat Prod Res. 30:1417–1422. 2016.PubMed/NCBI View Article : Google Scholar
8	Huang WH, Zhou GX, Wang GC, Chung HY, Ye WC and Li YL: A new biflavonoid with antiviral activity from the roots of Wikstroemia indica. J Asian Nat Prod Res. 14:401–406. 2012.PubMed/NCBI View Article : Google Scholar
9	Li J, Lu LY, Zeng LH, Zhang C, Hu JL and Li XR: A new C-3/C-3'-biflavanone from the roots of Wikstroemia indica. Molecules. 17:7792–7797. 2012.PubMed/NCBI View Article : Google Scholar
10	Shao M, Huang XJ, Sun XG, Wang Y, Yang Y, Wang QR, Fan Q and Ye WC: Phenolic constituents from rhizome of Wikstroemia indica and their anti-tumor activity. Nat Prod Res Dev. 26:851–855. 2014.
11	Guo GM, Li W and Wang Y: Sterol compounds from Wikstroemia indica (L.) C.A.May. J Mt Agric Biol. 31:77–79. 2012.
12	Guo GM, Wang Y, Li W and Hao XJ: Study on the chemical constituents from the light petroleum extracts of Wikstroemia indica. Sci Technol Eng. 14:188–190. 2014.
13	Chen C, Qu F, Wang J, Xia X, Wang J, Chen Z, Ma X, Wei S, Zhang Y, Li JY, et al: Antibacterial effects of different extracts from Wikstroemia indica on Escherichiacoli based on microcalorimetry coupled with agar dilution method. J Therm Anal Calorim. 123:1583–1590. 2016.
14	Lu CL, Zhu L, Piao JH and Jiang JG: Chemical compositions extracted from Wikstroemia indica and their multiple activities. Pharm Biol. 50:225–231. 2012.PubMed/NCBI View Article : Google Scholar
15	Ho WS, Li YL, Sun SS, Ooi VE and Xue JY: Antiviral activity of daphnoretin isolated from Wikstroemia indica. Phytother Res. 24:657–661. 2010.PubMed/NCBI View Article : Google Scholar
16	Yang ZY, Kan JT, Cheng ZY, Wang XL, Zhu YZ and Guo W: Daphnoretin-induced apoptosis in HeLa cells: A possible mitochondria-dependent pathway. Cytotechnology. 66:51–61. 2014.PubMed/NCBI View Article : Google Scholar
17	Jiang HF, Wu Z, Bai X, Zhang Y and He P: Effect of daphnoretin on the proliferation and apoptosis of A549 lung cancer cells in vitro. Oncol Lett. 8:1139–1142. 2014.PubMed/NCBI View Article : Google Scholar
18	Liu MX, Chen YG, Luo J, Wang JL, Sun H and Chen J: One case of severe erythema multiform drug eruption accompanied by drug-induced liver injury induced by wikstroemiae indicae radix tablets. Chin J Pharmacovigilance. 15:314–315, 317. 2018.
19	National Pharmacopoeia Committee: Pharmacopoeia of People's Republic of China. Part 1. People's Health Publishing House, Beijing, p16, 1978.
20	Feng G, Li W, He X, Li ZQ, Wang JK, Zheng CQ, Tian XF and Leng AB: Effects of different extracts of Miao medicine Wikstroemia indica root on hepatotoxicity in normal rats. Chinese J Exp Traditional Med Formulae. 23:96–102. 2017.
21	National Pharmacopoeia Commission: Pharmacopoeia of the People's Republic of China. Part 1. China Medical Science and Technology Press, Beijing, p248, 2015.
22	Wang JK, Li W, Guo JM, Gao YM, Zhang LY and Yang R: Influence of different processing methods on the total flavonoids content of Wikstroemia indica. J Guiyang Coll Traditional Chin Med. 33:17–19. 2011.
23	Shen MF, Li W, Wang JK, Lin C, Guo JM, Yang CF and Yang Y: The content of daphnoretin in different medicinal parts of Wikstroemia indica was determined by HPLC. J Med Pharm Chin Minorities. 18:65–67. 2012.
24	Zhang JJ, Xiong Y, Li W, Wang JK, Lin C, Wu N and Yang Q: Comparison of antibacterial and anti-inflammatory effect of unprocessed and processed products of indian stringbush root. Lishizhen Med Materia Med Res. 26:1118–1120. 2015.
25	Feng G, Li W, He X, Zheng C, Leng A and Tian X: Comparison of acute toxicity effects of ethanol extract from different processed products of Miao Medi-cine Wikstroemia indica on mice. China Pharmacy. 28:3536–3540. 2017.
26	Zhang J, Li W, Wang J, Lin C and Liu Y: Comparison of acute toxicity of extract of unprocessed indian atringbush root and its two different processed products. Zhongguo Zhong Yao Za Zhi. 36:1172–1174. 2011.PubMed/NCBI(In Chinese).
27	Feng G, Chen YL, Li W, Li LL, Wu ZG, Wu ZJ, Hai Y, Zhang SC, Zheng CQ, Liu CX and He X: Exploring the Q-marker of ‘sweat soaking method’ processed radix Wikstroemia indica: Based on the ‘effect-toxicity-chemicals’ study. Phytomedicine. 45:49–58. 2018.PubMed/NCBI View Article : Google Scholar
28	Zhang Z, Wang X, Wang J, Jia Z, Liu Y, Xie X, Wang C and Jia W: Metabonomics approach to assessing the metabolism variation and endoexogenous metabolic interaction of ginsenosides in cold stress rats. J Proteome Res. 15:1842–1852. 2016.PubMed/NCBI View Article : Google Scholar
29	Kanehisa M, Araki M, Goto S, Hattori M, Hirakawa M, Itoh M, Katayama T, Kawashima S, Okuda S, Tokimatsu T and Yamanishi Y: KEGG for linking genomes to life and the environment. Nucleic Acids Res. 36:D480–D484. 2008.PubMed/NCBI View Article : Google Scholar
30	West TP: Pyrimidine bases catabolism in Pseudomonas putida biotype B. Antonie Van Leeuwenhoek. 80:163–167. 2001.PubMed/NCBI View Article : Google Scholar
31	Guo PP, Wang J, Dong G, Wei D, Li M, Yang M and Kong L: NMR-based metabolomics approach to study the chronic toxicity of crude ricin from castor bean kernels on rats. Mol Biosyst. 10:2426–2440. 2014.PubMed/NCBI View Article : Google Scholar
32	Araujo TT, Barbosa Silva Pereira HA, Dionizio A, Sanchez CD, de Souza Carvalho T, de Silva Fernandes M and Rabelo Buzalaf MA: Changes in energy metabolism induced by fluoride: Insights from inside the mitochondria. Chemosphere. 236(124357)2019.PubMed/NCBI View Article : Google Scholar
33	Xu F, Liu Y, Zhao H, Yu K, Song M, Zhu Y and Li Y: Aluminum chloride caused liver dysfunction and mitochondrial energy metabolism disorder in rat. J Inorg Biochem. 174:55–62. 2017.PubMed/NCBI View Article : Google Scholar
34	Venediktova NI, Mashchenko OV, Talanov EY, Belosludtseva NV and Mironova GD: Energy metabolism and oxidative status of rat liver mitochondria in conditions of experimentally induced hyperthyroidism. Mitochondrion. 52:190–196. 2020.PubMed/NCBI View Article : Google Scholar
35	Buckalew VM Jr: Nephrolithiasis in renal tubular acidosis. J Urol. 141:731–737. 1989.PubMed/NCBI View Article : Google Scholar
36	Zhang H, Chen Y, Li Y and Wang T: Protective effect of polydatin on jejunal mucosal integrity, redox status, inflammatory response, and mitochondrial function in intrauterine growth-retarded weanling piglets. Oxid Med Cell Longev. 7178123:2020.PubMed/NCBI View Article : Google Scholar
37	Tasci I, Mas N, Mas MR, Tuncer M and Comert B: Ultrastructural changes in hepatocytes after taurine treatment in CCl₄ induced liver injury. World J Gastroenterol. 14:4897–4902. 2008.PubMed/NCBI View Article : Google Scholar
38	Romano TG, Schmidtbauer I, Silva FM, Pompilio CE, D'Albuquerque LA and Macedo E: Role of MELD score and serum creatinine as prognostic tools for the development of acute kidney injury after liver transplantation. PLoS One. 8(e64089)2013.PubMed/NCBI View Article : Google Scholar
39	Gartland KP, Bonner FW, Timbrell JA and Nicholson JK: Biochemical characterisation of para-aminophenol-induced nephrotoxic lesions in the F344 rat. Arch Toxicol. 63:97–106. 1989.PubMed/NCBI View Article : Google Scholar
40	Holmes E, Nicholson JK, Nicholls AW, Lindon JC, Connor SC, Polley S and Connelly J: The identification of novel biomarkers of renal toxicity using automatic data reduction techniques and PCA of proton NMR spectra of urine. Chemometr Intell Lab Syst. 44:245–255. 1998.
41	Buchman AL: The addition of choline to parenteral nutrition. Gastroenterology. 137:119–128. 2009.PubMed/NCBI View Article : Google Scholar
42	Griffin JL, Mann CJ, Scott J, Shoulders CC and Nicholson JK: Choline containing metabolites during cell transfection: An insight into magnetic resonance spectroscopy detectable changes. FEBS Lett. 509:263–266. 2001.PubMed/NCBI View Article : Google Scholar
43	Oliveira DT, Chaves-Filho AB, Yoshinaga MY, Paiva NC, Carneiro CM, Miyamoto S, Festuccia WT and Guerra-Sá R: Liver lipidome signature and metabolic pathways in nonalcoholic fatty liver disease induced by a high-sugar diet. J Nut Biochemistry. 87(108519)2021.PubMed/NCBI View Article : Google Scholar