| Objective:By preparing the rat model of chronic heart failure（CHF）,we observe the changes of heart and kidney function,renin-angiotensin-aldosterone system（RAAS）and the expression of renal aquaporin（AQPs）in rats with chronic heart failure（CHF）by Xinshuitong herbal water extract（XST）,to explore the therapeutic mechanism of XST on CHF.Methods:（1）Model establishment,grouping and administration:72 clean male Wistar rats were randomly divided into normal control group,and they were given an equal volume of normal saline by intraperitoneal injection.The remaining rats were injected with adriamycin（Adr）to prepare CHF model,1.25 mg·kg-1 was injected intraperitoneally twice a week.After 6 weeks of modeling,high resolution small animal ultrasound imaging was performed.The rats whose the left ventricular ejection fraction（LVEF）≤50%were randomly divided into 5 groups in cages,8 in each group:the captopril group was given 2.625 mg·（kg·d）-1captopril suspension by intragastric administration,XST low dose group is 4.95 g·（kg·d）-1,medium dose group is 9.9 g·（kg·d）-1,high dose group is 19.8g·（kg·d）-1),and intragastric administration in a volume of 10 mL·kg-1;the normal control group and the model group were given an equal volume of distilled water.Each group was intragastrically administered once a day for 4 weeks.After 4 weeks of administration,high resolution small animal ultrasound imaging was performed to detect changes in LVEF in rats.（2）Elisa measured serum renin（RI）,angiotensin II（Ang II）,aldosterone（ALD）content.（3）Automatic biochemical analyzer to detect rat serum and urine biochemical indicators.（4）The paraffin sections were taken from the kidney tissue,and the microstructure of the kidney was observed by HE staining.（5）Immunohistochemistry（IHC）,Western Blot and q-PCR were used to detect the morphological changes of kidney tissue and the genes and proteins expression of AQP1/2/3/4 in kidney.Result:1.XST treatment of CHF rats:（1）Ultrasound imaging of small animals showed LVEF results in rats:Compared with the normal control group,the LVEF of the model rats was significantly decreased（P<0.01）.After XST intervention,the LVEF in the middle and high dose groups of XST group was significantly different from that in the model group（P<0.05）.（2）The results of Elisa determination of RI,Ang II and ALD in rat serum showed that compared with the normal control group,the serum levels of RI,Ang II and ALD in the model group were significantly increased（P<0.01）.Compared with the model group,XST low,medium and high doses groups whose the level of RI,Ang II and ALD decreased in a dose-dependent manner,with significant difference（P<0.01）.（3）The results of automatic biochemical analyzer for measuring myocardial enzyme and liver and kidney function in rats showed that compared with the normal control group,the level of serum creatinine（Cr）,urea nitrogen（BUN）,creatine kinase（CK）,creatine kinase isoenzyme（CK-MB）,alanine aminotransferase（ALT）,aspartate aminotransferase（AST）was significantly increased（P<0.01）;Compared with the model group,the XST dose groups of Cr,BUN,CK,CK-MB,ALT and AST were all decreased in a dose-dependent manner,with significant difference（P<0.01）.At the same time,compared with the normal control group,there was no significant difference in blood and urine electrolyte levels between the XST dose group（P>0.05）.Compared with the model group,the rats’body weight of the XST dose groups decreased to different degrees,and the rats’food intake increased,and the difference was statistically significant（P<0.05）,but there was no difference in diuretic effect in the three dose groups of XST.（4）Renal histopathology observation results showed that compared with the normal control group,the renal tissue of the model group showed morphological changes,renal sac expanded,and the cavity is often filled with lightly stained protein fluid or a little red blood cells.Glomerular capillary vasospasmed,glomerular basement membrane vacuolar degenerated,renal tubular epithelial vacuolar degenerated,tubular atrophied.Some renal tubules are compensatory hypertrophy,occasionally protein cast.The morphological changes of kidney tissue in the drug-administered group were better than those in the model group,but some lesions of glomeruli and renal tubules were still seen.2.The effect of XST on the expression of AQP1/2/3/4 in kidney tissue:（1）The expression of AQP1 protein in kidney tissue detected by IHC and Western Blot showed that the expression of AQP1in rat kidney was significantly increased after model establishment（P<0.01）,and mainly distributed in the proximal tubule epithelial cell membrane and cytoplasm,mainly luminal membrane;after XST intervention,the expression of AQP1 in rat kidney tissue was down-regulated,the difference was statistically significant（P<0.05）.（2）The expression of AQP2 protein in kidney tissue detected by IHC and Western Blot showed that the expression of AQP2 in rat kidney was significantly increased after model establishment（P<0.01）,and it was mainly distributed in the plasma membrane of epithelial cells of collecting duct,mainly luminal membrane.After XST intervention,the expression of AQP2 in rat kidney tissue was down-regulated,and the difference was statistically significant（P<0.05）.（3）The expression of AQP3 protein in kidney tissue detected by IHC and Western Blot showed that the expression of AQP3 in rat kidney was significantly increased after model establishment（P<0.01）,and it was mainly distributed in the plasma membrane of epithelial cells of collecting duct,especially the basal side and membrane-based.After XST intervention,the expression of AQP3 in rat kidney tissue was down-regulated,and the difference was statistically significant（P<0.05）.（4）The expression of AQP4 protein in kidney tissue detected by IHC and Western Blot showed that the expression of AQP4 in rat kidney was significantly increased after model establishment（P<0.01）,and it was mainly distributed in the plasma membrane of epithelial cells of collecting duct,especially the basal side and membrane-based.Western Blot results showed that the expression of AQP4 in rat kidney tissue was down-regulated by XST intervention,and the difference was statistically significant（P<0.05）.（5）The relative expression of AQP1/2/3/4 mRNA in kidney tissue was detected by q-PCR.The results showed that the relative expression of AQP1/2/3/4 mRNA in rat kidney tissue was significantly increased after model establishment（P<0.01）.After XST intervention,the relative expression of AQP1/2/3/4 mRNA in kidney tissue was down-regulated,and the difference was statistically significant（P<0.05 or P<0.01）.Conclusion:Xinshuitong herbal water extract can improve heart functions and relieve water and sodium retention in chronic heart failure,which may be related to inhibition of RAAS activity and regulation of overexpression of renal AQPs. |
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