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Abstract：
BACKGROUND: Lagopsis supina (Steph.) Ik. -Gal. ex Knorr. has been widely used as a remedy treatment for diuresis and edema in China over 2500 years. Our previous results showed that the aqueous soluble fraction from L. supina (LSB) possessed acute diuretic effect.
OBJECTIVE: The aim of this study was to appraise the acute (6 h) and prolonged (7 d) diuretic effects, underlying mechanisms, and chemical profiling of LSB.
METHODS: The chemical profiling of LSB was performed by ultra-high-performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UHPLC-qTOF-MS/MS). Then, oral administration of LSB (40, 80, 160 and 320 mg/kg) and furosemide (10 mg/kg) once daily for 7 consecutive days to evaluate the diuretic effects in saline-loaded rats. The body weight, food consumption, and water intake were recorded once daily. The urinary volume, pH and electrolyte concentrations (Na+, K+, Cl-, and Ca2+) were measured after administration drugs for acute and prolonged diuretic effects. In addition, the serum levels of Na+-K+-ATPase, angiotensin II (Ang II), anti-diuretic hormone (ADH), aldosterone (ALD), atriopeptin (ANP), aquaporins (AQPs)-1, 2 and 3 were determined by ELISA kits. The mRNA expressions and protein levels of AQPs-1, 2 and 3 were analyzed by real-time quantitative PCR and Western blot assays, respectively.
RESULTS: 30 compounds were identified in LSB based on accurate mass and MS/MS fragmentation compared to literature, among which phenylpropanoids and flavonoids could be partly responsible for the major diuretic effect. Daily administration of LSB (160 or 320 mg/kg) prominently increased urinary excretion volume after the 2 h at the first day of treatment, remaining until the 7th day. LSB did not cause Na+ and K+ electrolyte abnormalities, and has minor effect on Cl- and Ca2+ concentrations at 320 mg/kg. Furthermore, LSB observably suppressed renin-angiotensin-aldosterone system (RAAS) activation, including decreased serum levels of Ang II, ADH, and ALD, and prominently increased serum level of ANP in rats. LSB treatment significantly down-regulated the serum levels, mRNA expressions and protein levels of AQP1, AQP2, and AQP3.
CONCLUSIONS: LSB has a prominent acute and prolonged diuretic effects via suppression of AQP and RAAS pathways in saline-loaded rats, and support the traditional folk use of this plant. Taken together, LSB might be a potential diuretic agent.
OBJECTIVE: The aim of this study was to appraise the acute (6 h) and prolonged (7 d) diuretic effects, underlying mechanisms, and chemical profiling of LSB.
METHODS: The chemical profiling of LSB was performed by ultra-high-performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UHPLC-qTOF-MS/MS). Then, oral administration of LSB (40, 80, 160 and 320 mg/kg) and furosemide (10 mg/kg) once daily for 7 consecutive days to evaluate the diuretic effects in saline-loaded rats. The body weight, food consumption, and water intake were recorded once daily. The urinary volume, pH and electrolyte concentrations (Na+, K+, Cl-, and Ca2+) were measured after administration drugs for acute and prolonged diuretic effects. In addition, the serum levels of Na+-K+-ATPase, angiotensin II (Ang II), anti-diuretic hormone (ADH), aldosterone (ALD), atriopeptin (ANP), aquaporins (AQPs)-1, 2 and 3 were determined by ELISA kits. The mRNA expressions and protein levels of AQPs-1, 2 and 3 were analyzed by real-time quantitative PCR and Western blot assays, respectively.
RESULTS: 30 compounds were identified in LSB based on accurate mass and MS/MS fragmentation compared to literature, among which phenylpropanoids and flavonoids could be partly responsible for the major diuretic effect. Daily administration of LSB (160 or 320 mg/kg) prominently increased urinary excretion volume after the 2 h at the first day of treatment, remaining until the 7th day. LSB did not cause Na+ and K+ electrolyte abnormalities, and has minor effect on Cl- and Ca2+ concentrations at 320 mg/kg. Furthermore, LSB observably suppressed renin-angiotensin-aldosterone system (RAAS) activation, including decreased serum levels of Ang II, ADH, and ALD, and prominently increased serum level of ANP in rats. LSB treatment significantly down-regulated the serum levels, mRNA expressions and protein levels of AQP1, AQP2, and AQP3.
CONCLUSIONS: LSB has a prominent acute and prolonged diuretic effects via suppression of AQP and RAAS pathways in saline-loaded rats, and support the traditional folk use of this plant. Taken together, LSB might be a potential diuretic agent.
摘要：
背景：Lagopsissupina(Steph.）Ik。-盖尔.前Knorr.2500多年来，在中国已被广泛用作利尿和水肿的补救措施。我们先前的结果表明，来自腹肌（LSB）的水溶性部分具有急性利尿作用。
目的：本研究的目的是评估急性（6h）和长期（7d）利尿作用，潜在机制，和LSB的化学分析。
方法：通过超高效液相色谱-四极杆飞行时间串联质谱（UHPLC-qTOF-MS/MS）进行LSB的化学谱分析。然后，连续7天每天一次口服LSB（40、80、160和320mg/kg）和呋塞米（10mg/kg），以评估盐水负荷大鼠的利尿作用。体重,食物消费,每天记录一次水摄入量。尿量,pH和电解质浓度（Na+，K+,Cl-,和Ca2）在给药急性和长期利尿作用后进行测量。此外,血清Na+-K+-ATP酶水平,血管紧张素II（AngII），抗利尿激素（ADH），醛固酮（ALD），心房肽(ANP),通过ELISA试剂盒测定水通道蛋白（AQPs）-1、2和3。通过实时定量PCR和Westernblot检测AQPs-1、2和3的mRNA表达和蛋白水平,分别。
结果：根据与文献相比的准确质量和MS/MS片段，在LSB中鉴定了30种化合物，其中苯丙素和类黄酮可能是主要利尿作用的部分原因。治疗第一天2小时后，每日服用LSB（160或320mg/kg）显着增加尿排泄量，一直到第七天。LSB没有引起Na+和K+电解质异常，在320mg/kg时对Cl-和Ca2+浓度影响较小。此外，LSB显著抑制肾素-血管紧张素-醛固酮系统（RAAS）激活，包括AngII的血清水平降低,ADH,和ALD,大鼠血清ANP水平显著升高。LSB治疗显著下调血清水平，AQP1、AQP2和AQP3的mRNA表达和蛋白水平。
结论：LSB在盐水负荷大鼠中通过抑制AQP和RAAS途径具有显著的急性和长期利尿作用，并支持这种植物的传统民间使用。一起来看,LSB可能是一种潜在的利尿剂。
目的：本研究的目的是评估急性（6h）和长期（7d）利尿作用，潜在机制，和LSB的化学分析。
方法：通过超高效液相色谱-四极杆飞行时间串联质谱（UHPLC-qTOF-MS/MS）进行LSB的化学谱分析。然后，连续7天每天一次口服LSB（40、80、160和320mg/kg）和呋塞米（10mg/kg），以评估盐水负荷大鼠的利尿作用。体重,食物消费,每天记录一次水摄入量。尿量,pH和电解质浓度（Na+，K+,Cl-,和Ca2）在给药急性和长期利尿作用后进行测量。此外,血清Na+-K+-ATP酶水平,血管紧张素II（AngII），抗利尿激素（ADH），醛固酮（ALD），心房肽(ANP),通过ELISA试剂盒测定水通道蛋白（AQPs）-1、2和3。通过实时定量PCR和Westernblot检测AQPs-1、2和3的mRNA表达和蛋白水平,分别。
结果：根据与文献相比的准确质量和MS/MS片段，在LSB中鉴定了30种化合物，其中苯丙素和类黄酮可能是主要利尿作用的部分原因。治疗第一天2小时后，每日服用LSB（160或320mg/kg）显着增加尿排泄量，一直到第七天。LSB没有引起Na+和K+电解质异常，在320mg/kg时对Cl-和Ca2+浓度影响较小。此外，LSB显著抑制肾素-血管紧张素-醛固酮系统（RAAS）激活，包括AngII的血清水平降低,ADH,和ALD,大鼠血清ANP水平显著升高。LSB治疗显著下调血清水平，AQP1、AQP2和AQP3的mRNA表达和蛋白水平。
结论：LSB在盐水负荷大鼠中通过抑制AQP和RAAS途径具有显著的急性和长期利尿作用，并支持这种植物的传统民间使用。一起来看,LSB可能是一种潜在的利尿剂。
