| ObjectiveIn this study,2.5% adenine suspension and unilateral ureteral ligation were used to establish CRF rat model.The expression of apq1-4 in lung,kidney and bladder was studied by "tonifying kidney qi" of Jingui Shenqi Pill.The regulation and mechanism of water metabolism were discussed,and the scientificity of "kidney dominating water" theory was evaluated.Objective to provide experimental basis for the treatment and intervention of Jinkui Shenqi Pill in chronic kidney disease.Methods1.Experimental study of Jingui Shenqi Pill on adenine induced chronic renal failure in ratsReplication of CRF rat model by intragastric administration of adenine: 60 healthy male SD rats were randomly divided into normal group(n=20)and adenine model group(n=40).After one week of adaptive feeding,the CRF rat model was reproduced by intragastric administration of 2.5% adenine suspension at a dose of 250 mg /(kg · d)for 4 weeks.10 rats in each group were taken for pathological evaluation of blood,urine and kidney tissues.Experimental study on the treatment of adenine CRF rats with Jingui Shenqi Pill: after the establishment of adenine CRF rat model,40 rats in Experiment 1 were divided into normal group,CRF model group,low-dose Jingui Shenqi Pill group and high-dose Jingui Shenqi Pill group,with 10 rats in each group.The low-dose and high-dose groups of Jingui Shenqi Pill were given corresponding concentration of Jingui Shenqi solution by gavage respectively.The rats in the normal group and CRF model group were given the same amount of distilled water by gavage for 4 weeks.The 24-hour urine protein,blood biochemical indexes and renal histopathology of each group were compared.2.Experimental study of Jingui Shenqi Pill on UUO rats with chronic renal failureEstablishment of CRF rat model by UUO method: 50 healthy male SD rats were randomly divided into sham operation group(n = 10)and UUO model group(n = 40).The CRF model was established by UUO operation.The rats were anesthetized by intraperitoneal injection of 10% chloral hydrate 350 mg / kg.The muscle and skin tissues were sutured at the lower part of the left kidney near the renal hilum and the upper 1 / 3 of the ureter.Rats in the sham operation group were given intraperitoneal anesthesia with 10% chloral hydrate for laparotomy,only the left ureter was blunt separated without ligation;rats in the normal group were not given any operation.Two weeks after operation,rats were randomly selected to evaluate the CRF model of UUO rats.Experimental study of Jingui Shenqi Pill on CRF in UUO rats: rats in each group were divided into normal group sham oper,ation group,UUO model group,low-dose group and high-dose group.Rats in normal group,sham operation group and UUO model group were given distilled water by gavage,while rats in low-dose and high-dose Jinkui Shenqi Pill treatment groups were given corresponding concentration of Jinkui Shenqi solution by gavage for 4 weeks.The general symptoms,24-hour urine protein,blood biochemical index and pathological changes of kidney tissue of rats in each group were compared.(the normal group was the same as experiment 1)3.Expression of aquaporin in rats with chronic renal failureAfter 4 weeks of treatment with adenine gavage and Jingui Shenqi Pill,the corresponding aquaporins AQP1,AQP2,AQP3 and AQP4 were detected by Western blotting in kidney,lung and bladder tissues of CRF rats.Image J software was used to analyze the gray value of different protein bands of AQP1-4 obtained by Western blotting on the kidney,lung and bladder of rats in each group.The dapdh protein bands were corrected.4.Statistical methods: SPSS18.0 software was used to analyze the experimental results,(x ±s)was used to measurement data analysis.Two independent samples t-test and And one-way ANOVA were used to compare the mean between groups.,Bilateral test,Inspection levelα= 0.05,P<0.05 was defined as statistically significant difference.Results1.24h-UTP,BUN and SCR of adenine model group were higher than those of normal group at 4 weeks(P < 0.05).The pathological changes of kidney were consistent with those of CRF.Compared with adenine model group,the levels of 24h-UTP,BUN and SCR in Jingui Shenqi Pill treatment group were significantly lower(P < 0.05).Compared with the adenine model group,the pathological damage of kidney in the treatment group of Jingui Shenqi Pill was alleviated in varying degrees.2.The levels of 24h-UTP,BUN and SCR in UUO group were significantly higher than those in normal group(P < 0.05).The pathological changes of kidney were consistent with those of CRF.Compared with UUO model group,the levels of 24h-UTP,BUN and SCR in each treatment group of Jingui Shenqi Pill were significantly lower(P < 0.05).Compared with the UUO model group,the renal pathological injury of rats was alleviated in varying degrees.3.AQP protein in kidney,lung and bladder of rats in each group was detected(1)The expression of AQP1-4 in kidney and AQP3 in bladder were up-regulated in adenine model group,which was significantly higher than that in normal group(P < 0.05).The expression of AQP1 in lung and bladder was significantly lower than that in normal group(P< 0.05).(2)The expression of AQP1 and AQP4 in kidney,AQP1 in lung and AQP4 in bladder in UUO model group were significantly lower than those in normal group(P < 0.05);The expression of AQP3 in bladder was up-regulated,and the difference was statistically significant compared with the normal group(P < 0.05).(3)Compared with adenine model group,AQP1-4 in kidney of high-dose adenine CRF Jingui Shenqi Pill treatment group was significantly decreased(P < 0.05).Compared with adenine model group,the AQP1 level of bladder was significantly increased(P <0.05).(4)Compared with UUO model group,the expression of AQP4 protein was significantly increased in high-dose group(P < 0.05).Conclusions1.Jingui Shenqi Pill can improve the quality of life of adenine and UUO rats,improve renal function,reduce 24-hour urinary protein,alleviate the pathological damage of kidney,especially improve the degree of renal fibrosis.2.Jingui Shenqi Pill can treat UUO and adenine CRF mainly by up regulating AQP levels of different types and parts.Adenine CRF mainly plays a role by down regulating renal AQP1-4 and up regulating bladder AQP1 level.The AQP2,AQP3 and AQP4 levels in kidney and lung of UUO rats were not significantly up-regulated,but the AQP4 protein level in bladder was significantly up-regulated. |
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