Experimental Observation On The Intervention Of Wuling Powder On Water Metabolism And Mechanisms In Model Rats With Kidney-yang Deficiency And Edema

ObjectiveTaking aquaporin 4 as the breakthrough point,we carried out the research on the treatment of abnormal water metabolism related diseases in rats with kidney-yang deficiency edema by Wuling San,a water-diversifying prescription;through the study of aquaporin,we may help to find a new method to improve the disorder of water metabolism and a new target of potential drug action,and to extract the mechanism of regulating water metabolism prescription in treating kidney-yang deficiency edema.For new research ideas.According to the instructor's many years clinical application of Wuling Powder in the treatment of water metabolism-related diseases,the effect of Wuling Powder on the changes of water metabolism,the activity of Na~+-K~+-ATP ase and the expression of aquaporin 4 in rats with kidney-yang deficiency edema was studied.MethodFirstly,50 SD rats were fed in metabolic cages in barrier environment for 3 days to adapt to the environment.Doxorubicin hydrochloride was injected into tail vein on the first and eighth day of the model.The two doses were 4 mg/kg and 3.5 mg/kg,respectively.Hydrocortisone was injected into arm muscles for 14 days,the dosage was 3.75 mg/kg,and the model was finished on 14 days.After the completion of the model,the successful rats were randomly divided into model group(M group),Wuling powder low dose group(W-L group),Wuling powder middle dose group(W-M group),Wuling powder high dose group(W-H group),furosemide group(F group),8 rats in each group.In addition,8 rats without intervention were selected as normal group(NG group).Group W-L,group W-M and group W-H were given Wuling powder crude drug 0.13g/kg,0.26g/kg and 0.52g/kg by body weight respectively,group F furosemide was given orally at 0.36mg/ml,2ml each time,twice one day.Group NG and group M were given saline of equal volume by gavage.Each group was given continuous gastric perfusion for 7 days.The urine was collected in metabolic cage for 24 hours.The urinary protein content was measured by BCA.At the end of the experiment,the kidneys of rats were taken out.Na~+-K~+-ATP ase activity of Kidney was measured by ultramicro ATP(Na~+-K~+-ATP ase)kit,kidney AQP4 concentration was detected by ELISA,and AQP4 protein expression and distribution weredetected by immunohistochemistry and Western Blot.The expression of AQP-4 protein in kidney tissue was detected and analyzed by real-time PCR.Result1.After the establishment of the model,rats suffered from weight loss,hypothermia,hair dryness,slow reaction,loss of appetite,incomplete stool,and abdominal and claw edema.24-hour urine volume decreased significantly,24-hour urinary protein content increased significantly.Referring to the diagnostic criteria of renal edema in traditional Chinese medicine and the literature of model building,combined with the above symptoms,the experiment was successful.2.It was found that after Wuling powder and furosemide treatment,urine volume increased,stool began to take shape,food intake,weight,body temperature increased,mental state improved,urine volume increased,edema gradually disappeared,and urinary protein decreased.In group F,the effect was faster and the recovery was better in group of W-M and W-H.3.After modeling,the activity of Na~+-K~+-ATP ase decreased and the expression of AQP4 in renal tissue increased.After Wuling powder intervention,the activity of Na~+-K~+-ATP ase increased and the expression of AQP4 in renal tissue decreased.4.After treatment with Wuling Powder,the activity of Na~+-K~+-ATP ase increased and the expression of AQP-4 decreased.Statistical analysis shows significant significance.Conclusion1.Wuling Powder has significant diuretic and edema-reducing effect in treating kidney-yang deficiency edema.Different concentrations have different curative effects.The middle and high doses of Wuling Powder are more ideal.2.After modeling,Na~+-K~+-ATP ase activity decreased and AQP4 expression increased in kidney tissue.After Wuling powder intervention,Na~+-K~+-ATP ase activity increased and AQP4 expression in kidney tissue decreased.3.Wuling Powder treatment of edema model rats with kidney-yang deficiency is related to the increase of Na~+-K~+-ATP ase activity and the down-regulation of AQP4 expression.