Search JIM Advanced Search

Journal of Chinese Integrative Medicine ›› 2007, Vol. 5 ›› Issue (3): 307-313.doi: 10.3736/jcim20070315

• Original Experimental Research • Previous Articles     Next Articles

Metabonomic study of syndrome of liver qi stagnation and spleen deficiency in rats

He-gu Luo, Jie Ding, Guang-xin Yue, Jia-xu Chen   

  1. Department of Diagnostics of Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing 100029, China
  • Online:2007-05-31 Published:2007-05-15

Objective: To determine the changes of the plasma metabolic phenotype in rats with chronic restraint stress (rats with syndrome of liver qi stagnation and spleen deficiency), so as to reveal the biological features of the syndrome of liver qi stagnation and spleen deficiency, and to examine potential application of metabonomic analysis in studies of syndromes in traditional Chinese medicine.Methods: Twenty-four male Sprague-Dawley rats were randomly divided into four groups: group A, 7 d normal control group; group B, 21 d normal control group; group C, 7 d stress group; and group D, 21 d stress group, with 6 rats in each group. Chronic restraint was used to induce stress in rats. Blood was collected from the cardio-ventricle under anesthesia on the 8th (groups A and C) or 22nd day (groups B and D) and detected by using the Fourier variable superconducting nuclear magnetic resonance (NMR) spectrometer (Varian UnityInova 600 M). Free induction decay signals were transferred into one-dimensional NMR spectrogram via 32 k Fourier transformation. Segmental integral calculus (0.04 ppm per segment) was performed from 4.5-0.5 ppm (Carr-Purcell-Meiboom-Gill, CPMG) or 6.0-0 ppm (longitudinal eddy-delay, LED) as defaulted 1H spectra values by using the VNMR software. Data were saved as text or excel files after normalization and then used for pattern recognition analyses. All the data were analyzed by principal component analysis (PCA) using the SIMCA-P 10.0 software (Umetrics AB, Umea, Sweden).Result: The PCA analysis of rat plasma 1H NMR spectra revealed different metabolic spectra between stress and control groups, which were consistent with alterations of in vivo metabolisms in rats under stress stimuli. Compared with the normal control group, rats with repeated stress displayed significant changes in spectral peak shapes of acetate, lactate, tyrosine, low-density lipoprotein, and unknown compounds (3.44 ppm). These altered metabolites can be used as biomarkers of syndrome of liver qi stagnation and spleen deficiency for further studies. Conclusion: The 1H NMR spectra of metabolites in the rat blood are differentially changed after chronic stress. Specific, characteristic metabolic products can be identified by analyses of metabonomics, which lead to interpretation of biological feature of Chinese medicine syndromes. Therefore, metabonomic analysis is an approach with good development prospects to studies of TCM syndromes.

Key words: metabonomics, pattern recognition, nuclear magnetic resonance, traditional Chinese medicine, rats

CLC Number: 

  • R25

Figure 1

Serum spectra of 600-M Hz 1H NMR in 4 groupsA: Group A (7 d normal control group); B: Group B (21 d normal control group); C: Group C (7 d stress group); D: Group D (21 d stress group)."

Figure 2

PCA score (t[4] vs t[5]) and loading (p[4] vs p[5]) plots for groups A and C serum spectra (LED)A: Group A (7 d normal control group); C: Group C (7 d stress group)."

Figure 3

PCA score (t[3] vs t[4]) and loading (p[3] vs p[4]) plots for groups B and D serum spectra (LED)B: Group B (21 d normal control group); D: Group D (21 d stress group)."

Figure 4

PCA score (t[1]vs t[2]) and loading (w*c[1] vs w*c[2]) plots for groups A and C serum spectra (CPMG) A: Group A (7 d normal control group); C: Group C (7 d stress group)."

Figure 5

PCA score (t[1] vs t[2]) and loading (p[1] vs p[2]) plots for groups A and B serum spectra (LED)A: Group A (7 d normal control group); B: Group B (21 d normal control group)."

Figure 6

PCA score (t[1] vs t[2]) and loading (p[1] vs p[2]) plots for groups C and D serum spectra (LED)C: Group C (7 d stress group); D: Group D (21 d stress group)."

Figure 7

PCA score (t[2] vs t[3]) and loading (p[2] vs p[3]) plots for groups A and B serum spectra (CPMG)A: Group A (7 d normal control group); B: Group B (21 d normal control group)."

Figure 8

PCA score (t[1] vs t[2]) and loading (p[1] vs p[2]) plots for groups C and D serum spectra (CPMG)C: Group C (7 d stress group); D: Group D (21 d stress group)."

[1] Waters NJ, Waterfield CJ, Farrant RD , et al. Metabonomic deconvolution of embedded toxicity: application to thioacetamide hepato- and nephrotoxicity[J]. Chem Res Toxicol, 2005,18(4):639-654
[2] Lindon JC, Nicholson JK, Holmes E , et al. Metabonomics: metabolic processes studied by NMR spectroscopy of biofluids[J]. Concepts Magn Reson, 2000,12(5):289-320
doi: 10.1002/1099-0534(2000)12:5<289::AID-CMR3>3.0.CO;2-W
[3] 陈家旭, 李伟, 赵歆 , 等. 慢性束缚应激大鼠海马脑啡肽mRNA和前强啡肽mRNA表达及中药复方的影响[J]. 中国应用生理学杂志, 2005,21(2):121-125
Chen JX, Li W, Zhao X , et al. Effects of Chinese herbs on enkaphalin mRNA and prodynorphin mRNA gene expression in rat hippocampus with chronic immobilization stress[J]. Zhongguo Ying Yong Sheng Li Xue Za Zhi, 2005,21(2):121-125
[4] 徐洪雁, 赵歆, 陈家旭 , 等. 慢性束缚应激大鼠海马CA1区L-ENK的变化及逍遥散的调节作用[J]. 世界科学技术——中医药现代化, 2005,7(5):18-22
Xu HY, Zhao X, Chen JX , et al. Changes of L-ENK in area CA1 of rats' hippocampus in condition of chronic immobilization stress and regulating actions of Xiaoyao powders[J]. Shi Jie Ke Xue Ji Shu–Zhong Yi Yao Xian Dai Hua, 2005,7(5):18-22
[5] 陈家旭, 杨建新, 赵歆 , 等. 慢性束缚应激大鼠下丘脑-β内啡肽变化及中药复方对其的影响[J]. 中国医药学报, 2004,19(2):83-85
Chen JX, Yang JX, Zhao X , et al. Changes of β-endorphine in rats' hypothalamus in condition of chronic immobilization stress and influence of Chinese herbal prescriptions[J]. Zhongguo Yi Yao Xue Bao, 2004,19(2):83-85
[6] 陈家旭, 唐已婷 . 逍遥散对慢性束缚应激模型大鼠相关脑区CRF基因表达的影响[J]. 中国应用生理学杂志, 2004,20(1):71-74
Chen JX, Tang YT . Effect of Xiaoyao powder on changes of relative brain zone CRF gene expression in chronic restrained stress rats[J]. Zhongguo Ying Yong Sheng Li Xue Za Zhi, 2004,20(1):71-74
[7] 陈家旭, 李伟, 赵歆 , 等. 三种中药复方对慢性束缚应激大鼠行为及皮层和海马NT3的影响[J]. 北京中医药大学学报, 2004,27(2):19-23
Chen JX, Li W, Zhao X , et al. Effects of three TCM formulas on the behavior and NT_3 in the cortex and hippocampus in the rats irritated by the chronic stressor of restraint[J]. Beijing Zhong Yi Yao Da Xue Xue Bao, 2004,27(2):19-23
[8] Chen JX, Li W, Zhao X , et al. Changes of mRNA expression of enkephalin and prodynorphin in hippocampus of rats with chronic immobilization stress[J]. World J Gastroenterol, 2004,10(17):2 547-2 549
[1] Wan-jun Guo, Yi Wang, Yu Deng, Lin-yan Cheng, Xin Liu, Ruo-fan Xi, Sheng-jie Zhu, Xin-yi Feng, Liang Hua, Kan Ze, Jian-yong Zhu, Dong-jie Guo, Fu-lun Li. Therapeutic effects of the extract of Sancao Formula, a Chinese herbal compound, on imiquimod-induced psoriasis via cysteine-rich protein 61. Journal of Integrative Medicine, 2022, 20(4): 376-384.
[2] Shang-jin Song, Xuan Liu, Qing Ji, Da-zhi Sun, Li-juan Xiu, Jing-yu Xu, Xiao-qiang Yue. Ziyin Huatan Recipe, a Chinese herbal compound, inhibits migration and invasion of gastric cancer by upregulating RUNX3 expression. Journal of Integrative Medicine, 2022, 20(4): 355-364.
[3] Marisa Casal. Improving the health and treatment success rates of in vitro fertilization patients with traditional chinese medicine: need for more robust evidence and innovative approaches. Journal of Integrative Medicine, 2022, 20(3): 187-192.
[4] Ning Guo, Fei Wu, Mei Wu, Yuan Wang, Qing Lang, Xiao Lin, Yi Feng. Progress in the design and quality control of placeboes for clinical trials of traditional Chinese medicine . Journal of Integrative Medicine, 2022, 20(3): 204-212.
[5] Yan-jun Lin, Kun-li Jiao, Bo Liu, Lu Fang, Shu Meng. Antiplatelet and myocardial protective effect of shexiang tongxin dropping pill in patients undergoing percutaneous coronary intervention: A randomized controlled trial. Journal of Integrative Medicine, 2022, 20(2): 126-134.
[6] Jia-jia Li, Qing Liang, Guang-chun Sun. Traditional Chinese medicine for prevention and treatment of hepatocellular carcinoma: A focus on epithelial-mesenchymal transition. Journal of Integrative Medicine, 2021, 19(6): 469-477.
[7] Rui-jin Qiu, Min Li, Jia-yuan Hu, Jing Chen, Hong-cai Shang. Methods for development of a core outcome set for clinical trials integrating traditional Chinese medicine and Western medicine. Journal of Integrative Medicine, 2021, 19(5): 389-394.
[8] Mao-xing Pan, Chui-yang Zheng, Yuan-jun Deng, Kai-rui Tang, Huan Nie, Ji-qian Xie, Dong-dong Liu, Gui-fang Tu, Qin-he Yang, Yu-pei Zhang. Hepatic protective effects of Shenling Baizhu powder, a herbal compound, against inflammatory damage via TLR4/NLRP3 signalling pathway in rats with nonalcoholic fatty liver disease . Journal of Integrative Medicine, 2021, 19(5): 428-438.
[9] Ning Zhang, Xiao-he Xiao. Integrative medicine in the era of cancer immunotherapy: Challenges and opportunities. Journal of Integrative Medicine, 2021, 19(4): 291-294.
[10] Thomas Efferth, An-long Xu, Roxana Damiescu, Mita Banerjee, Norbert W. Paul, David Y.W. Lee. Can eastern wisdom resolve western epidemics? Traditional chinese medicine therapies and the opioid crisis. Journal of Integrative Medicine, 2021, 19(4): 295-299.
[11] Si-qi Tang, Yun-liang Wang, Zi-ye Xie, Yang Zhang, Yi Guo, Kang-li Gao, Tang-you Mao, Chun-e Xie, Jun-xiang Li, Xiao-yan Gao. Serum metabolic profiling of traditional Chinese medicine syndromes in patients with diarrhea-predominant irritable bowel syndrome. Journal of Integrative Medicine, 2021, 19(3): 274-281.
[12] Miao-yan Shi, Shi-qi Sun, Wei Zhang, Xing Zhang, Gui-hua Xu, Xuan Chen, Zi-jian Su, Xiu-ming Song, Lu-jiong Liu, Yi-bao Zhang, Yi-le Zhang, Meng Sun, Qi Chen, Yan Xue, Hua Lü, Wei-an Yuan, Xiao-rong Chen, Yun-fei Lu. Early therapeutic interventions of traditional Chinese medicine in COVID-19 patients: a retrospective cohort study. Journal of Integrative Medicine, 2021, 19(3): 226-231.
[13] Shu-jun Wei, Qing-man He, Qing Zhang, Kang-hua Fu, Ruo-lan Li, Wei Peng, Yong-xiang Gao. Traditional Chinese medicine is a useful and promising alternative strategy for treatment of Sjogren's syndrome: A review. Journal of Integrative Medicine, 2021, 19(3): 191-202.
[14] Yi Xia, Lu-shao-bo Shi, Jing-hui Chang, Hua-zhang Miao, Dong Wang. Impact of the COVID-19 pandemic on intention to use traditional Chinese medicine: A cross-sectional study based on the theory of planned behavior. Journal of Integrative Medicine, 2021, 19(3): 219-225.
[15] Na Zhi, Qian Mo, Shuo Yang, Yuan-xing Qin, Hao Chen, Zeng-guang Wu, Cai-hong Lan, Jun Zhang, Yin-long Li. Treatment of pulmonary fibrosis in one convalescent patient with corona virus disease 2019 by oral traditional Chinese medicine decoction: A case report. Journal of Integrative Medicine, 2021, 19(2): 185-190.
Full text



[1] Dong Yang, Yong-ping Du, Qing Shen, Wei Chen, Yan Yu, Guang-lei Chen. Expression of alpha-smooth muscle actin in renal tubulointerstitium in patients with kidney collateral stasis. Journal of Chinese Integrative Medicine, 2008, 6(1): 41-44
[2] Hai-feng Wei, Bai-liu Ya, Ling Zhao, Cui-fei Ye, Li Zhang, Lin Li. Evaluation of tongue manifestation of blood stasis syndrome and its relationship with blood rheological disorder in a rat model of transient brain ischemia. Journal of Chinese Integrative Medicine, 2008, 6(1): 73-76
[3] Xi Lin, Jian-ping Liu. Herbal medicines for viral myocarditis. Journal of Chinese Integrative Medicine, 2008, 6(1): 76
[4] Xi Lin, Jian-ping Liu. Tai chi for treating rheumatoid arthritis. Journal of Chinese Integrative Medicine, 2008, 6(1): 82
[5] Liang-ping Hu, Hui Gao. Discrimination of errors in statistical analysis of medical papers published in the first issue of 2006 in Journal of Chinese Integrative Medicine. Journal of Chinese Integrative Medicine, 2008, 6(1): 98-106
[6] Yan-bo Zhu , Qi Wang, Cheng-yu Wu, Guo-ming Pang, Jian-xiong Zhao, Shi-lin Shen, Zhong-yuan Xia , Xue Yan . Logistic regression analysis on relationships between traditional Chinese medicine constitutional types and overweight or obesity. Journal of Chinese Integrative Medicine, 2010, 8(11): 1023-1035
[7] Wei Xu, Meng Shi, Jian-gang Liu, Cheng-long Wang . Collagen protein expressions in ischemic myocardium of rats with acute myocardial infarction and effects of qi-tonifying, yin-tonifying and blood-activating herbs and detoxifying and blood-activating herbs. Journal of Chinese Integrative Medicine, 2010, 8(11): 1041-1047
[8] Tao Wang , Feng Qin. Effects of Chinese herbal medicine Xiaoyao Powder on monoamine neurotransmitters in hippocampus of rats with postpartum depression. Journal of Chinese Integrative Medicine, 2010, 8(11): 1075-1079
[9] Ying Xu , Chang-chun Zeng , Xiu-yu Cai , Rong-ping Guo , Guang Nie , Ying Jin. Chromaticity and optical spectrum colorimetry of the tongue color in different syndromes of primary hepatic carcinoma. Journal of Chinese Integrative Medicine, 2012, 10(11): 1263-1271
[10] Xiang-ying Mao , Qin Bian , Zi-yin Shen. Analysis of the osteogenetic effects exerted on mesenchymal stem cell strain C3H10T1/2 by icariin via MAPK signaling pathway in vitro. Journal of Chinese Integrative Medicine, 2012, 10(11): 1272-1278