甘遂半夏汤中甘遂-甘草反药组合加减对腹水大鼠利水药效及RAAS系统的影响

Diuretic effect and impact on RAAS system of the ascites model rats by Gansui Banxia Decoction with or without Kansui Radix-Licorice incompatible combination

目的:探究甘遂半夏汤中甘遂-甘草反药组合加减治疗腹水大鼠的利水药效及其机制。方法:雄性Wistar大鼠随机分为空白组、模型组、阳性药组、甘遂半夏汤全方组(全方组)、甘遂半夏汤全方去生甘遂组(去遂组)、甘遂半夏汤全方去炙甘草组(去草组)、甘遂半夏汤全方去生甘遂炙甘草组(双去组)。腹腔注射Walker-256细胞复制大鼠腹水模型。造模后第2天给药,第3天收集24 h尿液,记录尿量,检测尿液上清液中Na^(+)、K^(+)、Cl^(-)含量;第9天选取造模成功大鼠,腹主动脉取血后摘取双侧肾脏,检测血清Na^(+)、K^(+)、Cl^(-)含量,血浆肾素活性(PRA)、血管紧张素Ⅱ(AngⅡ)、醛固酮(ALD)、抗利尿激素(ADH)含量及肾脏水通道蛋白2(AQP2)mRNA和蛋白表达。结果:与空白组比较,模型组尿量显著减少(P<0.01),血浆PRA、AngⅡ、ALD、ADH含量显著升高(P<0.01),肾脏AQP2 mRNA和蛋白表达显著升高(P<0.01,P<0.05),尿Na^(+)量显著降低(P<0.01),血清K^(+)、Cl^(-)量显著升高(P<0.01)。与模型组比较,阳性药组、全方组尿量显著增多(P<0.01,P<0.05),血浆PRA、AngⅡ、ALD、ADH含量显著降低(P<0.01);全方组肾脏AQP2 mRNA和蛋白表达显著降低(P<0.05);全方组尿Na^(+)量显著升高(P<0.01),阳性药组、全方组血清K^(+)、Cl^(-)量显著降低(P<0.01,P<0.05)。与全方组比较,去草组和双去组尿量显著减少(P<0.01,P<0.05),去遂组、去草组和双去组血浆PRA、AngⅡ、ALD、ADH显著升高(P<0.01),去遂组和去草组肾脏AQP2 mRNA表达显著升高(P<0.01,P<0.05),去草组和双去组尿Na^(+)量显著降低(P<0.01),双去组血清K^(+)、Cl^(-)量显著升高(P<0.01)。结论:甘遂半夏汤全方及全方加减一味或两味反药组合可增加腹水大鼠尿量,其中全方利水药效佳,属于“增效”配伍关系,其“增效”机制一方面可能通过增强全方对RAAS系统的抑制;另一方面可能通过增加全方对肾脏AQP2 mRNA和蛋白表达的抑制。通过以上两方面甘遂半夏汤全方对水液的重吸收减少,体液排出增多,利水药效增加,并且改善了腹水引起的血尿电解质紊乱。

Objective:To explore the Kansui Radix-Licorice incompatible combination’s diuretic effect and underlying mechanism by taking Gansui Banxia Decoction with or without Kansui Radix-Licorice incompatible combination to the ascites model rats.Methods:Male Wistar rats were randomly divided into the normal control group,model group,positive control group,whole formula group,Gansui removed group,honey-roasted Licorice removed group,double removed group.The ascites rat model was induced by intraperitoneal injection of Walker-256 tumor cells.The drug was administered the day after making model,and three days later,24 h urine was collected.Then testing the level of Na^(+),K^(+),and Cl^(-)in urine supernatant.On the 9th day of taking drug,collecting abdominal aorta blood and bilateral kidneys from the successful established model rats.Then testing the serum Na^(+),K^(+),Cl^(-)contents,the plasma PRA,AngⅡ,ALD,ADH levels,the protein and mRNA expression of AQP2 in kidney.Results:Compared with the normal control group,the urine volume was significantly reduced(P<0.01),the plasma PRA,AngⅡ,ALD,and ADH were significantly higher(P<0.01),the renal AQP2 mRNA and protein expression were significantly higher(P<0.01,P<0.05),the urinary Na^(+)amount was significantly lower(P<0.01),the serum K^(+)and Cl^(-)amount was significantly higher(P<0.01)in the model group.Compared with the model group,the urine volume was significantly increased(P<0.01,P<0.05),the plasma PRA,AngⅡ,ALD,and ADH levels were significantly lower in the positive control group and the whole formula group(P<0.01);the renal AQP2 mRNA and protein expression were significantly lower(P<0.05)in the whole formula group;the urinary Na^(+)amount was significantly higher(P<0.01)in the whole formula group,the serum K^(+)and Cl^(-)amount were significantly lower(P<0.01,P<0.05)in the positive control group and the whole formula group.Compared with the whole formula group,the urine volume was significantly reduced(P<0.01,P<0.05)in the honey-roasted Licorice removed group and the double removed group;the plasma PRA,AngⅡ,ALD,and ADH were significantly higher(P<0.01)in the gansui removed group,the honey-roasted Licorice removed group and the double removed group;the renal AQP2 mRNA expression was significantly higher(P<0.01,P<0.05)in the Gansui removed group and the honey-roasted Licorice removed group;the urinary Na^(+)amounts were significantly lower(P<0.01)in the honey-roasted Licorice removed group and the double removed group;the serum K^(+)and Cl^(-)amounts were significantly higher(P<0.01)in the double removed group.Conclusion:By taking Gansui Banxia Decoction with or without Kansui Radix-Licorice incompatible combination,the ascites rats’urine volume is increase.The whole formula group’s diuretic effect is the best,therefore the Kansui Radix-Licorice incompatible combination could increase its diuretic efficacy.The mechanism of Kansui Radix-Licorice incompatible combination increasing Gansui Banxia Decoction’s diuretic efficacy may be related to two aspects:on the one hand,by inhibiting RAAS system,on the other hand,by increasing the inhibition of the mRNA and protein expression of AQP2 in renal.Through the above two aspects,the whole formula group reduces the reabsorption of water,increases the drainage of body fluid,therefore enhances the diuretic efficacy,and improves the blood and urine electrolyte disorders caused by ascites.