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J Zhejiang Univ (Med Sci)  2018, Vol. 47 Issue (6): 628-635    DOI: 10.3785/j.issn.1008-9292.2018.12.11
    
Effects of Niaoduqing granule on urine metabolic profile in chronic renal failure rats
ZHU Min1(),WU Yunqiu2,SHOU Zhangfei1,*()
1. Department of Nephrology, Shulan Hangzhou Hospital, Hangzhou 310004, China
2. Jiangsu Province Academy of Traditional Chinese Medicine, Nanjing 210028, China
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Abstract  

Objective: To investigate the effects of Niaoduqing granule on the urine metabolic profile in chronic renal failure (CRF) rats. Methods: Thirty six male SD rats were divided into the normal control group, the model group, and the Niaoduqing group with 12 rats in each group. The CRF was induced by gavage of 250 mg·kg-1·d-1 adenine for 21 d. UPLC-Q-TOF-MS/MS technique was used in combination with principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) to analyze the urine metabolic profiles in three groups. The endogenous substances with the variable importance projection (VIP)>1 and P < 0.05 were screened as the potential biomarkers for CRF, and enrichment analysis of metabolic pathways was carried out. Results: Compared with the normal control group, the model group had lower body weight, higher kidney coefficient, higher serum creatinine and urea nitrogen levels (all P < 0.01), while the above indexes in the Niaoduqing group were ameliorated compared with the model group (all P < 0.01). Fifteen potential biomarkers were found in the urine of the model group, which were involved in 9 metabolic pathways including phenylalanine, tyrosine and tryptophan biosynthesis, glyoxylate and dicarboxylate metabolism, valine, leucine and isoleucine biosynthesis, arachidonic acid metabolism, cysteine and methionine metabolism, tricarboxylic acid cycle, glycerophosphatide metabolism, tryptophan metabolism and tyrosine metabolism. Conclusion: Niaoduqing granules has therapeutic effect on rats with CRF, which may be related to the regulation of amino acid metabolism, lipid metabolism and energy metabolism.



Key wordsKidney failure, chronic      Drugs, Chinese herbal/therapeutic use      Urine      Metabolism      Biological markers      Chromatography, liquid      Mass spectrometry      Disease models, animal     
Received: 01 June 2018      Published: 15 March 2019
CLC:  R917  
  R692  
Corresponding Authors: SHOU Zhangfei     E-mail: min.zhu@shulan.com;zhangfei.shou@shulan.com
Cite this article:

ZHU Min,WU Yunqiu,SHOU Zhangfei. Effects of Niaoduqing granule on urine metabolic profile in chronic renal failure rats. J Zhejiang Univ (Med Sci), 2018, 47(6): 628-635.

URL:

http://www.zjujournals.com/med/10.3785/j.issn.1008-9292.2018.12.11     OR     http://www.zjujournals.com/med/Y2018/V47/I6/628


基于尿液代谢组学技术研究尿毒清颗粒治疗慢性肾功能衰竭的机制

目的: 运用代谢组学方法研究尿毒清对慢性肾功能衰竭(CRF)大鼠尿液代谢谱的影响,并探讨其机制。方法: 36只大鼠随机分为空白对照组、模型对照组和尿毒清组。采用腺嘌呤法制备大鼠CRF模型。运用超高效液相色谱串联四极杆飞行时间质谱技术结合主成分分析和偏最小二乘判别分析,比较空白对照组、模型对照组、尿毒清组大鼠尿液的代谢谱,筛选出变量重要性投影值>1且P < 0.05的内源性物质作为CRF潜在的生物学标志物,并进行代谢通路的富集分析。结果: 与空白对照组比较,模型对照组大鼠体质量降低、肾脏系数增高、血清肌酐和尿素氮水平升高(均P < 0.01),而尿毒清组上述指标较模型对照组均有所改善(均P < 0.01)。共鉴定出15个潜在生物学标志物,涉及9条代谢通路紊乱,分别为苯丙氨酸、酪氨酸、色氨酸的生物合成,乙醛酸和二羧酸代谢,缬氨酸、亮氨酸、异亮氨酸生物合成,花生四烯酸代谢,半胱氨酸和蛋氨酸代谢,三羧酸循环,甘油磷脂代谢,色氨酸代谢,酪氨酸代谢。结论: 尿毒清对CRF大鼠具有一定的治疗作用,其机制可能与调节氨基酸代谢、脂质代谢和能量代谢等有关。


关键词: 肾功能衰竭, 慢性,  中草药/治疗应用,  尿,  代谢,  生物学标记,  色谱法, 液相,  质谱分析法,  疾病模型, 动物 
($\bar x \pm s$)
组别 n 体质量(g) 肾脏系数(%) 肌酐(μmol/L) 尿素氮(μmol/L)
造模0 d 造模21 d 造模31 d
与空白对照组比较,**P < 0.01;与模型对照组比较,#P < 0.05, ##P < 0.01.
空白对照组 12 215±24 274±30 310±40 1.12±0.11 40±5 12±3
模型对照组 12 225±36 248±26** 266±31** 3.04±0.47** 96±10** 48±7**
尿毒清组 12 219±31 242±25** 280±29**# 1.77±0.36**## 62±8**## 27±5**##
Tab 1 The body weights, kidney coefficients, serum creatinine and urine nitrogen levels in three groups
Fig 1 Total ion chromatograms of urine samples from three groups
Fig 2 PCA score of normal control, model and Niaoduqing granules groups
保留时间(min) 生物学标志物 HMDB编号 KEGG编号 相对分子质量 分子式 VIP 趋势*
*模型对照组较空白对照组变化的趋势.HMDB:人类代谢组数据库;KEGG:京都基因与基因组百科全书;VIP:变量投影重要性指标.
14.27 酪氨酸 HMDB00158 C00082 181.07 C9H11NO3 4.31 升高
11.86 蛋氨酸 HMDB00696 C00073 149.05 C5H11NO2S 3.18 降低
13.51 色氨酸 HMDB00929 C00078 204.09 C11H12N2O2 3.18 升高
1.26 肌酸 HMDB00064 C00300 131.07 C4H9N3O2 2.41 升高
14.93 亮氨酸 HMDB00687 C00123 131.09 C6H13NO2 2.40 升高
20.50 花生四烯酸 HMDB01043 C00219 304.24 C20H32O2 1.97 升高
0.89 柠檬酸 HMDB00094 C00158 192.03 C6H8O7 1.61 降低
0.73 肌酐 HMDB00562 C00791 113.12 C4H7N3O 1.44 降低
14.05 半胱氨酸 HMDB00574 C00097 121.02 C3H7NO2S 1.43 降低
4.82 胆碱 HMDB00097 C00114 104.11 C5H14NO 1.26 升高
11.41 马尿酸 HMDB00714 C01586 179.06 C9H9NO3 1.19 降低
12.58 十二烷酸 HMDB00638 C02679 200.18 C12H24O2 1.14 升高
0.95 草酰乙酸 HMDB00223 C00036 132.01 C4H4O5 1.07 降低
20.33 1,2-二十六烷基 HMDB00564 C00157 733.56 C40H80NO8P 1.05 升高
21.47 棕榈酸 HMDB00220 C00249 256.24 C16H32O2 1.02 升高
Tab 2 Potential biomarkers of chronic renal failure in rats
($\bar x \pm s$)
组别 n 酪氨酸 蛋氨酸 色氨酸 肌酸 亮氨酸 花生四烯酸 柠檬酸 肌酐 半胱氨酸 胆碱 马尿酸 十二烷酸 草酰乙酸 1,2-二十六烷基 棕榈酸
与空白对照组比较,*P < 0.05, * *P < 0.01;与模型对照组比较,#P < 0.05, ##P < 0.01.
空白对照组 12 3.24±2.09 11.86±5.91 11.84±4.85 0.44±0.25 1.89±0.76 1.13±0.24 1.86±0.95 1.67±0.86 2.40±0.48 1.11±0.53 0.87±0.42 1.54±0.17 8.54±0.75 1.32±0.55 0.28±0.12
模型对照组 12 12.98±0.69** 4.55±3.42** 17.88±0.94** 3.38±0.18** 4.71±0.92* 3.68±1.63* 0.22±0.22** 0.21±0.20** 1.20±0.29* 2.02±0.15* 0.36±0.10* 2.36±0.43* 0.48±0.32** 2.03±0.21* 0.82±0.05*
尿毒清组 12 7.12±1.91## 4.04±2.03 12.86±1.88## 1.01±0.54# 2.39±1.02# 2.90±1.13 0.80±0.25# 0.53±0.17# 1.92±0.25# 1.65±0.30 0.94±0.20# 2.23±0.25 1.15±0.07# 1.45±0.20# 0.52±0.15
F 38.24 3.75 62.43 12.22 15.04 20.18 4.96 4.07 87.05 29.84 17.31 92.50 31.48 28.35 9.44
P < 0.01 < 0.05 < 0.01 < 0.01 < 0.01 < 0.01 < 0.05 < 0.05 < 0.01 < 0.01 < 0.01 < 0.01 < 0.01 < 0.01 < 0.01
Tab 3 Relative peak areas of potential biomarkers in three groups
Fig 3 Summary of pathway analysis
Fig 4 The metabolic network of urine endogenous metabolites of chronic renal failure rats
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