| Acute renal failure is a common clinical disease in dogs.It is characterized by a short course of disease,acute onset,and mostly complications,which seriously endangers the life and health of dogs.Immunity and other effects are well known,and in recent years,it has become a research hotspot of dietary nutritional additives and health care products.In this experiment,a canine acute renal failure model was established by adenine induction,to study the effect of nano-selenium on dogs with acute renal failure,to explore the preventive effect of nano-selenium on acute renal failure in dogs,and to provide new therapeutic ideas for the treatment of acute renal failure in dogs.1.Establishment of adenine-induced acute renal failure model in dogs:Fifteen Chinese garden dogs about 1 year old and weighing about 4kg were selected and randomly divided into blank control group,40mg/kg/d model group and75mg/kg/d model group.All three groups were fed basal diet and tested.Serum creatinine,blood urea nitrogen,urine specific gravity and urine pH on the 1st,8th,and15 th days,and the changes of kidney size were examined by imaging before the start of the experiment and after the end of the 15 th day.The experimental results showed that the dogs in the model group whose diet was supplemented with 75 mg/kg/d adenine had a significant increase in serum creatinine and blood urea nitrogen on the15 th day compared with the blank control group(P<0.05),and the serum creatinine level reached grade 3 renal failure.Standard(180-440 μmol/d L),urine pH and urine specific gravity were also significantly lower(P<0.05).Observation of renal histopathological sections in 75mg/kg/d adenine model group showed obvious swelling of epithelial cells,dilated lumens,and infiltration of inflammatory cells in the interstitium.In the imaging examination,the kidney size of the 75mg/kg/d adenine model group was larger than the normal standard range of 3.5×L2(second lumbar vertebra)compared with the blank control group.The high-dose group could stably achieve the renal failure index at 15 d,so 75mg/kg/d adenine was a reasonable dose for modeling renal failure in dogs.2.The effect of nano-selenium on the renal WNK-SPAK/OSR1 pathway in the course of adenine-induced acute renal failure in dogs and its protective effect on tissue: Twenty Chinese pastoral dogs about 1 year old and weighing about 4 kg were selected and randomly divided into modeling group,blank control group,fluid replacement treatment group,nano-selenium treatment group,and nano-selenium prevention group.Each group was fed the basal diet every day,and the modeling group,the rehydration treatment group,and the nano-selenium treatment group were fed with 75 mg/kg/d adenine on the basis of the diet for 15 days;after that,the modeling group continued to be fed the basal day.The 15-day dietary supplementation treatment group was supplemented with glucose and sodium chloride injection at the dose of the experimental dog’s body weight(60 ml/kg/d)for 15 days,while the nano-selenium treatment group was supplemented with nano-selenium 0.5mg/kg/d in the basal diet for 15 days.The nano-selenium prevention group was supplemented with 0.5 mg/kg/d of nano-selenium for 15 days,and then continued to be supplemented with 75 mg/kg/d of adenine for 15 days.At the end of 30 days,the experimental dogs were venous blood collection,bladder puncture to collect urine,and then euthanized by anesthesia to collect kidney tissue samples.The experimental results showed that compared with the control group,the urine specific gravity,urine pH and serum calcium concentration of the modeling group decreased,while the CRE,blood phosphorus and BUN concentrations increased.Decreases CRE,BUN and serum phosphorus levels.Immunohistochemical results showed that the expression of NKCC2 protein in the modeling group was significantly increased,while the expression of NKCC2 in the treatment group decreased,and the inflammatory infiltration was improved.RT-qPCR results showed that the expression of CaSR mRNA in the nano-selenium treatment group was significantly increased(P<0.05)and the expression of NKCC2 mRNA was significantly decreased(P<0.01)in the nano-selenium treatment group(P<0.01).Compared with the modeling group,the expression of Caspase3 mRNA was significantly increased(P<0.01),the expression of Caspase3 mRNA was significantly decreased(P<0.05),and the expression of Caspase9 mRNA and BAX mRNA was significantly decreased compared with the modeling group(P<0.05).The expression of-2 mRNA was significantly increased compared with the modeling group(P<0.05).Western-Blot results showed that the protein expression of WNK1 in the nano-selenium treatment group was significantly lower than that in the modeling group(P<0.05),and the expression of NKCC2 protein was significantly decreased(P<0.05).Compared with the modeling group,the expression level was significantly increased(P<0.05),the protein expression of Pro-Caspase3 was significantly decreased compared with the modeling group(P<0.05),and the protein of Pro-Caspase9 in the nano-selenium treatment group was significantly increased compared with the modeling group(P<0.05).);BAX protein expression was significantly decreased(P<0.05),and BCL-2 protein expression was significantly increased(P<0.01).Through the above experiments,it can be concluded that nano-selenium may improve calcium and phosphorus metabolism in the body,and indirectly reduce the ion transporter overexpressed in renal tissue due to the occurrence of acute renal failure through the CaSR-WNK/SPAK-NKCC2 pathway,thereby regulating the reabsorption function of the kidney.,Nano-selenium may improve the body’s calcium metabolism and reduce the expression of pro-apoptotic proteins in renal tissue through the CaSR-WNK-Caspase3 pathway to alleviate renal injury. |
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