作者：闫宇涵1，王占黎2，胡海3，刘秀芬4

单位：

1.014040 内蒙古包头市，包头医学院研究生学院　2.014040 内蒙古包头市，包头医学院转化医学中心 　3.014040 内蒙古包头市， 包头医学院基础医学与法医学院 　4.014040 内蒙古包头市，包头医学院人文社科与外国语学院

Units：

1.Graduate School, Baotou Medical College, Baotou 014040, China 2.Translational Medicine Center, Baotou Medical College, Baotou 014040, China 3.College of Basic Medicine and Forensic Medicine, Baotou Medical College, Baotou 014040, China 4.College of Humanities and Social Sciences and Foreign Languages, Baotou Medical College, Baotou 014040, China

关键词：

肾性高血压；两肾一夹；气相色谱- 飞行时间质谱联用；代谢组学；大鼠

Keywords：

Renal hypertensions; 2K1C; GC-TOFMS; Metabonomics; Rats

CLC：

R 544.14

DOI：

10.12114/j.issn.1008-5971.2021.00.151

Funds：

国家自然科学基金资助项目（81660048）

摘要：

背景　研究表明，粪便内源性代谢物可能与高血压的发病有关。但目前关于高血压特别是肾性高血压 大鼠肠道粪便内源性代谢物的研究鲜有报道。目的　基于气相色谱- 飞行时间质谱联用（GC-TOFMS）技术分析肾性 高血压大鼠模型粪便代谢组学。方法　本实验时间为2019-08-15至2019-09-30。选取4周龄SPF级SD雄性大鼠20只， 随机分为假手术组（n=10）和模型组（n=10）。模型组大鼠构建两肾一夹（2K1C）肾性高血压大鼠模型，假手术组 大鼠打开腹腔后进行左肾动脉分离但不夹闭。术后4 周处死大鼠并观察其肾脏体积；比较两组大鼠术后即刻及术后4 周尾动脉收缩压及体质量；基于GC-TOFMS 技术及多元变量统计分析方法筛选两组大鼠差异粪便代谢物，并进行层次 聚类分析。结果　模型组大鼠均建模成功。实验过程中模型组1 只大鼠死亡。术后4 周，与假手术组大鼠相比，模型 组大鼠左侧肾脏体积明显缩小、且形态萎缩。术后4 周模型组大鼠尾动脉收缩压高于假手术组，体质量低于假手术组 （P ＜ 0.05）。术后4 周模型组大鼠尾动脉收缩压高于术后即刻（P ＜ 0.05）；术后4 周假手术组和模型组大鼠体质 量分别高于本组术后即刻（P ＜ 0.05）。基于GC-TOFMS 技术和多元变量统计分析方法，共筛选出11 种差异粪便代 谢物：与假手术组相比，模型组葡萄糖酸、肌醇、壬酸、癸酸呈低表达，葡萄糖-1- 磷酸、麦芽三糖醇、异麦芽酮糖、 马来酸、2- 甲基戊二酸、2- 单棕榈酸甘油酯、L- 犬尿氨酸呈高表达。结论　肾性高血压大鼠粪便代谢物与假手术组 明显不同，本研究初步证明了肾性高血压对肠道内源性代谢物的影响。

Abstract：

Background　Studies have shown that fecal endogenous metabolites may be related to the pathogenesis of hypertension. However, there are few reports about the endogenous metabolites in the feces of hypertensive rats, especially renal hypertensive rats. Objective　To analyze the fecal metabonomics in renal hypertensive rats based on gas chromatography-timeof- flight mass spectrometry (GC-TOFMS) technology. Methods　The experiment lasted from August 15, 2019 to September 30, 2019. Twenty 4-week-old male SD rats of SPF grade were selected and randomly divided into Sham group (n=10) and model group (n=10). 2K1C renal hypertension rat model was established in the model group, and the left renal artery was separated but not clamped in the Sham group. The rats were killed 4 weeks after operation and the size of kidney was observed. The systolic blood pressure of tail artery and body mass were compared between the two groups immediately after operation and 4 weeks after operation; based on GC-TOFMS technology and multivariate statistical analysis, the different fecal metabolites of the two groups were screened and analyzed by hierarchical cluster analysis. Results　All rats in the model group were successfully established. One rat died of model group during the experiment. At 4 weeks after operation, compared with Sham group, the left kidney of model group was significantly reduced in volume and morphology. At 4 weeks after operation, the systolic blood pressure of tail artery of model group was higher than that of Sham group, and the body weight of model group was lower than that of Sham group (P < 0.05) . At 4 weeks after operation, the systolic blood pressure of tail artery in model group was higher than that immediately after operation (P < 0.05) ; the body weight in Sham group and model group at 4 weeks after operation were higher than those immediately after operation, respectively (P < 0.05) . Based on GC-TOFMS technology and multivariate statistical analysis, a total of 11 different feces metabolites were screened out: compared with the Sham group, the expression of glucose, inositol, nonanoic acid and decanoic acid in model group was low, while the expression of glucose-1-phosphate, maltotriitol, palatinose, maleamate, 2-methylglutaric acid, 2-monopalmitin and L-kynurenine in model group was high. Conclusion　The faecal metabolites of renal hypertensive rats were significantly different from those of Sham group, which preliminarily proved the effect of renal hypertension on intestinal endogenous metabolites.

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