| Yinhuamiyanling Tablets is a compounded Chinese medicinal prescription with tens of medicinal materials including honeysuckle.Some researches on diuretic efficacy have been finished on Polygonum aviculare,fringed pink,pyrrosia lingua,caulis clematidis armandii,semen plantaginis,lophatherum gracile,junci medulla and some other medicinal materials.But clinically confined to the treatment of acute renal nephritis,acute cystitis,coke damp heat syndrome and other urinary tract infections.The study group used modern pharmacological techniques to study the bacteriostatic and anti-inflammatory effects of Yinhuamiyanling Tablets,has not yet confirmed whether it has diuretic effect.Based on the previous research results and the feedback of clinical data,the pharmacokinetic test was carried out on the diuretic effect of Yinhuamiyanling Tablets,and the diuretic mechanism was discussed.The aim of this study was to provide reference for the further clinical application.In this study,the classical rat saline load model was used to study the diuretic effect of Yinhuamiyanling Tablets and its mechanism.The rats were randomly divided into normal saline group,low,medium and high dose group,and furosemide positive control group.After 8 days of continuous gavage,the test tube was placed under the metabolic cage and the urine was recorded once a hour.The urine was collected for 6 hours.The levels of p H,creatinine,Na~+,K~+ and Cl~- in the plasma / urine were measured and the diuretic effect was evaluated.After the experiment was completed,the serum of the rats was isolated and the serum was detected by Enzyme-linked immunosorbent assay(ELISA)(P <0.05).The expression of AQP1,AQP2,AQP3 and AQP4 m RNA in rat kidney were measured by real-time PCR.The morphological changes of kidney were evaluated by HE staining.The distribution and expression of AQP1,AQP2,AQP3 and AQP4 were observed by immunohistochemistry.The results showed that the urine level(P <0.001)was significantly increased in the low,medium and high dose groups of Yinhuamiyanling Tablets after 8 days of continuous administration,and the diuretic effect of Yinhuamiyanling Tablets high dose group was slightly stronger than that of fen(P <0.05).The contents of Na~+,K~+,Cl~- ion and creatinine in plasma were significantly higher than those in saline group(P> 0.05)(P <0.01).There was no significant difference in the levels of creatinine(P <0.01)between the two groups(P <0.01).The results of ELISA showed that the levels of RA,Ang? and ALD in Yinhuamiyanling Tablets group were significantly lower than those in saline group Real-time PCR results showed that the AQP4 m RNA levels in the drug groups were not significantly different from those in the saline group(P> 0.05),while the expression levels of AQP1,AQP2 and AQP3 were significantly higher than those in the control group(P <0.05,P <0.01).The results of HE staining showed that compared with the saline group,with the increase of the drug concentration,the renal tubular structure of the drug group showed a certain degree of expansion,the renal cortex and myeloid The expression of AQP4 in renal cortex was not significantly different from that in the control group(P <0.05).Compared with the saline group,the expression of AQP4 in the renal cortex was significantly lower than that in the normal saline group AQP1,AQP2,AQP3 protein expression level(P <0.01),which AQP2 decreased the most significant(P <0.001).In summary,we found that Yinhuamiyanling Tablets has an obvious diuretic effect.Our findings preliminarily demonstrate its mechanism in the renin-angiotensin-aldosterone(RAAS)system,reduce RA,Ang ? and ALD secretion,by inhibiting of renal medulla AQP1,AQP2,AQP3 protein expression levels,thereby affecting the renal tubular and collecting ducts on the role of the original urine reabsorption so as to play diuretic effect. |
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