| Objective:Chronic renal failure(CRF)is the end result of various chronic kidney diseases.The common pathological characteristic is renal interstitial fibrosis.The chronic renal failure model was established by continuous intragastric administration of adenine in rats,and observe the effect of Shenshuai Jiangzhuo Recipe on renal tissue of adenine rat model.The body weight,kidney weight index,serum creatinine,urea nitrogen,24 hour urinary protein quantity,aspartate aminotransferase,alanine aminotransferases,renal pathology,Wnt /β-catenin signal pathway and the expression of related protein and m RNA were measured 14 days after administration.Based on the Wnt/β-catenin signaling pathway,we observed the effect of different doses of Shenshuai Jiangzhuo Decoction on the renal tissues of adenine rats,and explored the mechanism of anti-fibrosis and prevention and treatment of chronic renal failure.Research method:Sixty healthy male Wistar rats were randomly divided into 6 groups: control group,model group,western medicine group,low,middle and high dose groups of Chinese medicine,10 rats in each group.Except the control group,rats in 5 groups were given continuous adenine(concentration of 30 mg / m L)by gavage for 3 weeks,The control group was given equal volume of normal saline,200 mg / kg once.After successful modeling,rats in each group were treated by gavage.The control group and model group were given normal saline2 ml / day,and the western medicine group was treated with 0.046 g / ml losartan potassium solution 2m L / day.The low,medium and high group of traditional Chinese medicine were given shenshuaijiangzhuo formula 2ml / day with solubility of 0.048g/ml,0.096g/ml and0.192g/ml,once a day for 14 days.After 14 days of treatment,serum and 24 h urine were collected from rats in each group,and biochemical indexes such as Bun,Scr,24 h urine protein quantification,ALT and AST could be detected.The left kidney was removed,weighed,and segmented.One part of the kidney was used for pathological examination and the other part for Western blot and RT-PCR.Research result:1.Effects of rh on general condition and biochemical indexes in ratsAfter intragastric administration of adenine,rats showed varying degrees of reduced food intake,reduced sensitivity to external stimuli,weight loss,decreased freedom of movement,increasingly depressed spirit,haggard and loose fur.After treatment,the weight gain rate of rats in each group was lower than that in the control group,and the serum urea nitrogen and creatinine had a certain increase to a certain extent.The final research results of the experiment showed that the rats in the control group had good indicators,while the model group had the worst indexes.The indexes in the treatment group were high-dose Chinese medicine group,western medicine group> Chinese medicine medium-dose group> Chinese medicine low-dose group.From the rat body weight,the weight of rats in each group have increased,in the adenine intragastric administration of rats increased slowly than the control group,During the treatment period,the body weight of the rats in the control group increased the fastest,and the model group increased the slowest.After 14 days of treatment,compared with the model group,there were significant differences between the the traditional Chinese medicine,high,medium,and low dose groups and the western medicine group.It shows that the drug treatment is effective,and the high-dose Chinese medicine group and the western medicine group have the most significant effect.Biochemical indexes: After adenine administration,serum urea nitrogen and creatinine increased in all groups except the control group.After 14 days of treatment,compared with the model group,the two index values of each administration group decreased.The decreasing effect of the low-dose group of traditional Chinese medicine was not obvious,and the high-dose group of traditional Chinese medicine and the western medicine group decreased significantly,with statistical difference.Compared with the control group,the ALT index of the model group decreased slightly and the AST level increased slightly.There was no significant difference between the groups.Compared with the model group,there was no significant difference in the levels of ALT and AST in the treatment group.2.Pathological effects on rat kidney tissueThe microscopic changes of renal tissue can be observed more intuitively by observing the pathological sections of the rat kidneys with HE and Masson staining.After adenine administration,the kidneys became larger.HE staining showed that the number of glomeruli in the model group decreased significantly and the balloon adhesion appeared;Atrophy and expansion appeared in the lumen of renal tubules,with inflammatory cells,protein tubules,vacuolar degeneration and exfoliated epithelial cells,with brown crystals in the tubes;Glomerular sclerosis,mesangial matrix hyperplasia and basement membrane thickening;Cell proliferation was obvious,fibroblasts and collagen fibers increased,and renal interstitial fibrosis was obvious.Compared with the model group,the number of glomeruli increased,the pathological changes of renal tissue were reduced,the renal interstitial fibrosis and inflammatory infiltration were reduced,and the high-dose group of traditional Chinese medicine was the most obvious.Masson staining showed that collagen fibers in renal tissue accumulated significantly and stained significantly,filling the whole renal interstitium.After administration,the blue staining area in renal interstitium in each group was less than that in the model group,and the collagen deposition was also reduced,especially in the high-dose group of traditional Chinese medicine.It proved that the fibrosis process was improved after administration.3.The effects on Wnt/β-catenin signaling pathway and RIF phenotype index protein expressionThe results of Western blot showed that in the blank group,there were a small amount of Wnt4 and β-catenin proteins in kidney tissue,and Wnt4 and β-catenin proteins were significantly increased after adenine gavage,indicating that they were related to the occurrence of chronic renal failure.After 14 days of treatment with western medicine and traditional Chinese medicine,the levels of Wnt4 and β-catenin in the group were decreased.Among them,The effect of high dose group was the most obvious.It is proved that the high-dose group of traditional Chinese medicine is the most effective for the disease.The expression of α-SMA in the kidney tissue of the rats in the model group was increased,while the expression of E-cadherin protein,which should have been highly expressed,was decreased.After treatment,the protein expression of α-SMA in each treatment group was decreased,the expression of E-cadherin protein expression was also increased in every treatment group,and the expression was the highest in the high-dose group.It was highly expressed after adenine gavage in TGF-β1 experiments.After the treatment with traditional Chinese medicine and western medicine,the protein expression of each treatment group decreased to varying degrees,among which the high-dose group of traditional Chinese medicine had the best curative effect,indicating that the high-dose group of traditional Chinese medicine had a therapeutic effect on chronic renal failure.Analysis conclusion:1.Shenshuai Jiangzhuo Decoction can effectively improve the general condition,increase body weight,decrease serum urea nitrogen,creatinine,24 h urinary protein quantitative and improve renal function in adenine rat model,high-dose Chinese medicine and western medicine group had the best effect.2.Shenshuai Jiangzhuo Decoction can improve the pathological changes caused by adenine rat model,and the effect is best in the high-dose group of Chinese medicine.3.Shenshuai jiangzhuo decoction can regulate Wnt4 / β-catenin signal pathway,Decreased the expression of Wnt4,β-catenin,α-SMA and TGF-β1 proteins and m RNA transcription levels,and increased the expressions of E-cadherin protein and m RNA transcript levels,The high dose group of Chinese medicine has the best effect.Reducing excessive deposition of collagen fibrils and extracellular matrix and alleviating the transdifferentiation of renal tubular epithelial cells to mesenchymal,thus resisting renal fibrosis and preventing the progression of chronic renal failure.4.Shenshuai Jiangzhuo Decoction can inhibit the renal pathological changes induced by adenine in rats,and the most obvious effect is in high dose group of traditional Chinese medicine. |
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