杜仲-刺蒺藜对老龄自发性高血压大鼠肠道微生物组的影响

Effects of Eucommia ulmoides-Tribulus terrestris on intestinal microbiota in aged spontaneously hypertensive rats

  • 摘要: 目的观察杜仲-刺蒺藜对自发性高血压大鼠(SHR)的血压及肠道微生物组的影响。方法选用18月龄雄性SHR 28只,将其随机分为对照组(SHR)和杜仲-刺蒺藜干预高、中、低剂量组(H、M、L)共4组,灌胃给药8周,每周测体质量、血压。8周后,酶联免疫吸附试验(ELISA)法检测血清氨基末端脑钠尿肽前体(NT-proBNP)、神经肽Y、C反应蛋白(CRP)、血管紧张素Ⅱ(AngⅡ)、去甲肾上腺素(NE)水平;实时荧光定量聚合酶链反应(qRT-PCR)法检测胸主动脉IκB激酶(IKK)α、IKK-β、核转录因子κB抑制蛋白(IκB)α、核转录因子(NF)-κB p65的mRNA表达水平;无菌原则取大鼠应激粪便样本,16S V4区扩增子测序与分析;采用气相色谱法检测粪便样本中短链脂肪酸(SCFA)的含量。结果高、中剂量杜仲-刺蒺藜干预后,大鼠血压明显低于SHR对照组收缩压/舒张压:(170.3±10.7)/(114.0±6.6)、(171.6±10.4)/(112.4±7.3)比(203.8±11.4)/(136.8±8.0)mm Hg,均P<0.05;与对照组比较,杜仲-刺蒺藜中剂量组血清NT-proBNP(321.5±31.4)比(471.7±61.9) ng/L、神经肽Y(2001.9±26.6)比(3826.7±68.4)ng/L、CRP(42.4±13.0)比(68.5±9.5)μg/mL、AngⅡ(481.9±21.4)比(591.6±88.7)ng/L水平显著降低(均P<0.05),杜仲-刺蒺藜高剂量组血清神经肽Y水平(2977.6±64.5)比(3826.7±68.4)ng/L,P<0.05显著降低。高、中剂量杜仲-刺蒺藜显著改善胸主动脉中IκBα、IKKβ、NF-κB p65、IKKα的mRNA表达水平(均P<0.05)。药物干预后大鼠肠道微生物的组成、α多样性及β多样性发生改变,Anaeroplasma为杜仲-刺蒺藜中剂量组在属水平上的特征菌属。高、中剂量杜仲-刺蒺藜显著升高大鼠粪便乙酸、丙酸水平;低剂量杜仲-刺蒺藜显著升高大鼠粪便丁酸、异丁酸、戊酸、异戊酸水平(均P<0.05);Anaeroplasma丰度与CRP浓度(r=-0.85)、NF-κB p65水平(r=-0.78)相关(均P<0.05)。结论杜仲-刺蒺藜可降低老龄SHR血压、降低炎症水平,其降压机制与肠道菌群的组成和多样性的改变相关。

     

    Abstract: Objective To observe the effects of the bark of eucommia(Eucommia ulmoides Oliv.) and tribulus terrestris(Tribulus terrestris L.)(ET) on blood pressure and intestinal microbiota in spontaneously hypertensive rats(SHR). Methods A total of 28 18-month-old male SHR were randomly divided into control group, ET high, medium and low dose group. Rats in four groups were gavaged for 8 weeks. Blood pressure and body weight were measured weekly. Eight weeks later, serum level of N-terminal pro brain natriuretic peptide(NT-proBNP), neuropeptide Y(NPY), C reactive protein(CRP), angiotensin Ⅱ(AngⅡ) and norepinephrine(NE) were measured by enzyme-linked immunosorbent assay(ELISA), and the expression levels of IKK-α, IKK-β, IκBα and NF-κB p65 were detected by real-time fluorescence quantitative PCR(qRT-PCR). Stool samples were taken from rats under the aseptic operation principle, and the amplicon information of 16 S V4 region was collected and analyzed. The content of short chain fatty acids(SCFA) in stool samples was detected by gas chromatography. Results The blood pressure of rats in ET high and medium dose group were significantly lower than that of SHR in control group systolic blood pressure/diastolic blood pressure:(170.3±10.7)/(114.0±6.6),(171.6±10.4)/(112.4±7.3) vs(203.8±11.4)/(136.8±8.0)mm Hg, P<0.05. Compared with the control group, medium dose of ET significantly decreased the levels of NT-proBNP, NPY, CRP and AngⅡ NT-proBNP:(321.5±31.4) vs(471.7±61.9)ng/L; NPY:(2001.9±26.6) vs(3826.7±68.4)ng/L, CRP:(42.4±13.0) vs(68.5±9.5)mg/L, AngⅡ:(481.9±21.4) vs(591.6±88.7)ng/L, P<0.05, high dose of ET significantly decreased NPY level (2977.6±64.5) vs(3826.7±68.4)ng/L, P<0.05. High and medium dose of ET can significantly change the expression of IκBα, IKKβ, NF-κB p65 and IKKα in thoracic aorta. The composition, alpha diversity and beta diversity of intestinal microbiota were significantly different after ET treatment. Anaeroplasma was characteristic microbiota of ET medium dose group at genus level. High and medium dose ET significantly increased the levels of acetic acid and propionic acid in stool samples, while low dose ET significantly increased the levels of butyric acid, isobutyric acid, valeric acid and isovaleric acid. The abundance of anaeroplasma was linearly correlated with CRP concentration(r=-0.85) and NF-κB p65 level(r=-0.78). Conclusion ET effectively reduced blood pressure and inflammation level in elderly SHR. The mechanism of antihypertension effect may be related to the changes of composition and diversity of intestinal microbiota.

     

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