甘遂与甘草配伍对癌性腹水大鼠AQP2/ADRB1/PIK3CG/AVPR2通路的调节作用

Regulatory effect of the AQP2/ADRB1/PIK3CG/AVPR2 pathway in rats with cancer ascites by compatibility of Radix Euphorbiae Kansui and Radix Glycyrrhizae

目的:探讨甘遂与甘草配伍对癌性腹水大鼠水盐代谢通路的调节作用。方法:将70只SD大鼠按体质量随机分为正常组、模型组、甘遂(0.7 g)组、甘草(10 g)组、配伍(甘遂0.7 g-甘草10 g)组、甘遂半夏汤(甘遂0.7 g、甘草10 g、半夏9 g、白芍15 g、蜂蜜15 g)组,其中正常组10只,其余每组12只。其中模型组和各给药组腹腔注射Walker-256细胞株,注射细胞株后第2天给予相应药物治疗,连续14 d。于第13天采用代谢笼收集尿液,以检测尿液中钠离子(Na;)、钾离子(K;)、β2微球蛋白(β2-MG)、肾损伤因子(KIM-1)、中性粒细胞明胶酶相关脂质运载蛋白(NGAL)的含量。给药14 d后,收集血清,检测各组血清肌酐(Cr)、尿素氮(BUN)、尿酸(UA)等肾功能指标以及肾素(Renin)、血管紧张素(ANG)Ⅱ、醛固酮(ALD)、抗利尿激素(ADH)水平。收集肾组织检测水通道蛋白2(AQP2)、β1肾上腺素能受体(ADRB1)、磷脂酰肌醇(-3)激酶γ亚基(PIK3CG)、精氨酸加压素受体2(AVPR2)mRNA及蛋白表达。结果:与模型组比较,甘遂与甘草配伍能降低血、尿β2-MG及尿液KIM-1、NGAL的含量(P<0.05),升高尿液中Na;含量,降低K;含量,降低血清Renin、ANGⅡ、ALD、ADH含量(P<0.05),下调肾AQP2、ADRB1、PIK3CG、AVPR2 mRNA和蛋白的表达(P<0.05)。结论:甘遂与甘草配伍能够调节水液代谢,其作用机制一方面是通过下调PIK3CG和ADRB的表达,从而降低AVPR2的表达;另一方面,通过降低ALD含量,从而影响K;的排出发挥作用。

Objective: To investigate the regulatory effect of the combination of Radix Euphorbiae Kansui and Radix Glycyrrhizae on water-salt metabolism pathway in rats with cancerous ascites. Methods: Seventy SD rats were randomly divided into normal group, model group, Radix Euphorbiae Kansui group(0.7 g), Radix Glycyrrhizae group(10 g), Radix Euphorbiae Kansui-Radix Glycyrrhizae group(0.7 g:10 g), Gansui Banxia Decoction group(Radix Euphorbiae Kansui 0.7 g, Radix Glycyrrhizae 10 g, Rhizoma Pinelliae 9 g, Radix Paeoniae Alba 15 g, Mel 15 g). There were 10 rats in normal group and 12 rats in each other group. The Walker-256 cell line was injected intraperitoneally in the model group and each treatment group, and the corresponding drug treatment was given on the second day after injection for 14 consecutive days. On the 13th day, urine was collected by metabolic cage to detect the contents of Na;, K;, β2-MG, KIM-1, NGAL. Forteen days after administration,serum was collected to detect renal function indexes such as Cr, BUN and uric acid(UA), as well as Renin, ANGⅡ, ALD and ADH levels. Renal tissues were collected to detect the mRNA and protein expression of AQP2, ADRB1, PIK3CG and AVPR2.Results: Compared with model group, the combination of Radix Euphorbiae Kansui and Radix Glycyrrhizae could decrease β2-MG in serum and urine, KIM-1, NGAL in urine(P<0.05), increase the content of Na;, decrease the content of K;, and decrease the contents of Renin, ANG2, ALD and ADH in serum(P<0.05), and down-regulate the mRNA and protein expression of AQP2, ADRB1, PIK3CG and AVPR2 in renal tissues(P<0.05). Conclusion: The combination of Radix Euphorbiae Kansui and Radix Glycyrrhizae can regulate water-liquid metabolism. On the one hand, the mechanism is to reduce the expression of AVPR2 by down-regulating the expression of PIK3CG and ADRB;On the other hand, by reducing the content of aldosterone, thus affecting the excretion of potassium ions play a role.