| ObjectiveTo observe the protective effect of rhubarb combined with aconite in Dahuang fuzi decoction on renal function in rats with unilateral ureteral obstruction and to explore its mechanism.MethodsSPF male Sprague-Dawley rats were divided into blank group,model group,2:1 group,1:1 group,1:2 group(compatibility ratio of rhubarb aconite was 2:1 、 1:1 、 1:2,the dosage of asarum was the same in each group)and positive drug group.The renal failure model was established by the left ureteral obstruction method.The intragastric administrated with Dahuang fuzi decoction and positive drug group.The general state of the rats was observed,urine was collected,and urine volume was recorded for the detection of 24-hour Urine protein quantification(24h UPQ).The rats were collected serum for detection of biochemical indicators(serum creatinine and serum urea nitrogen).Kidney tissues were collected,and the pathological changes of the kidney were observed by HE and Masson staining,and the expression and distribution of Notch1,PTEN and Hes1 proteins were detected by immunohistochemistry.Real-time PCR was used to detect the transcription levels of Notch1 m RNA,PTEN m RNA and Hes1 m RNA,and Western Blot method was used to detect the expression levels of Notch1,PTEN and Hes1 proteins.Result1.Dahuang fuzi decoction in each group could improve the pathological status of renal tissue injury in UUO model rats,and they were superior to the model group and the positive group in terms of mental state,skin color,food intake,body weight and urine volume,of which the 2:1 group had the most obvious effect.2.Dahuang fuzi decoction in each group could reduce the urinary proteincontent,serum creatinine and serum urea nitrogen levels in UUO model rats for 24 hours,with the most obvious effect in group 2:1.3.Dahuang fuzi decoction in each group could improve the degree of renal pathological changes in UUO model rats and reduce the percentage of renal fibrosis area,with the most obvious effect in group 2:1.4.Immunohistochemical results show: compared with the blank group,the expressions of Notch1 and Hes1 proteins in the kidney tissues of the model group were significantly increased,and the expressions of PTEN proteins were decreased(P<0.05).The expressions of Notch1 and Hes1 proteins in the kidney tissues of the treatment group were lower than that of the model group,and the expressions of PTEN proteins were higher than that of the model group(P<0.05).The effect was most obvious in the 2:1 group.5.Real-time PCR results show: the transcription levels of Notch1 and Hes1 m RNA in the kidney tissues of rats in the model group were significantly higher than those in the blank group,while the transcription levels of PTEN m RNA were lower(P<0.05).Compare with the model group,the transcription levels of Notch1 and Hes1 m RNA in the renal tissues of rats were down-regulated and the transcription levels of PTEN m RNA were up-regulated in the treatment group(P<0.05).The effect was most obvious in the 2:1 group.6.Western Blot results show:the expression of Notch1 and Hes1 proteins in kidney tissues of rats in the model group was significantly increased compared with those in the blank group,while the expression of PTEN proteins was decreased(P<0.05).Compare with the model group,the treatment group could down-regulate the expression of Notch1 and Hes1 proteins and up-regulate the expression of PTEN proteins in the renal tissues of rats(P<0.05).The effect was most obvious in the 2:1 group.Conclusion1.Dahuang fuzi decoction can improve the pathological status of UUO model rats,reduce the urine protein content,serum creatinine and serum ureanitrogen levels for 24 hours,and improve the pathological changes of kidney caused by ureteral obstruction.2.The best combination of rhubarb and aconite in Dahuang fuzi decoction is 2:1.3.Dahuang fuzi decoction may regulate the level of Notch signaling pathway by down-regulating Notch1,Hes1 and upregulation of PTEN,thus delaying renal interstitial fibrosis and protecting renal function. |
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