| Objective:To observe the effects of Professor Gao Jining's clinical experience formula "Yishen fang" on the expression of serum fibroblast growth factor 23(FGF23),Klotho mRNA in renal tissue and TGF-?1 factor in renal tissue of rats with early chronic renal failure(CRF)Mechanism to expand early medication selection for CRF.Methods:Rats were randomly divided into blank control,model control,Niaoduqing positive,Yishenfang low,medium and high dose,15 rats in each group.Except the blank control group,the other groups were established by adenine gavage in the early CRF rat model.After 4 weeks of modeling,rats were collected for 24 hours of urine,recorded urine volume,urine microalbumin(UmAlb)and 24 hours urine protein quantification(UTP);2rats were taken from each group,and serum creatinine(SCr)and urea nitrogen(BUN)and other blood biochemical indicators,selected based on results.After successful modeling,Yishen Decoction in low,middle and high dose groups were administered with gavage of0.8 g /(kg · d),1.6 g /(kg · d),3.2 g /(kg · d)with different doses,The Qing-positive control group was orally administered 1.6 g /(kg · d)of Niaoduqing granules,while the blank control group and the model control group were orally administered an equal volume of normal saline once a day for 8 weeks.Except for the blank control group,the other groups were given a 200 mg / kg adenine suspension at the same time,once every other day.The blank control group was given an equal amount of normal saline for 4 weeks.During dosing,changes in body weight and general conditions were recorded.After the administration,the blood and urine biochemical indicators were measured,and the hematoxylin and eosin staining(HE)and Masson staining(Massson)were used to observe the pathological changes of renal tissues.ELISA kits,reverse transcription polymerase chain reaction(RT-PCR))Method and Western Blot method were used to detect the levels of rat serum fibroblast growth factor FGF23,Klotho mRNA in renal tissue and TGF-?1factor in renal tissue.Results:1.General situation: After 4 weeks of modeling,the rats in the blank control group arein good condition,have agile movements,increase their body weight uniformly,and have a smooth coat color and luster.The rest of the groups were debilitated,sluggish,eating less water,reduced body mass,curled hair,yellow hair,thinner stools,and increased urine output.After 8 weeks of administration,compared with the model control group,the mental,diet,and defecation conditions of the rats in each group improved,especially in the high-dose group of Yishenfang.2.Changes in urine biochemical indicatorsAfter 4 weeks of modeling,except for the blank control group,the 24 h urine volume,UmAlb,and 24 h UTP of the remaining 4 groups were not significantly different from the model control group(P>0.05).After 8 weeks of administration,the model control group was significantly larger.The 24 h urine volume,UmAlb,and 24 hUTP of the rats were significantly higher than those of the blank control group,and the urine creatinine(UCr)was significantly lower than that of the blank control group(P<0.01);The above indexes are significantly lower than the model control group except UCr(P<0.05),and the above indexes of Yishenfang high-dose group are closer to the standard(P<0.01);urine N-acetyl-?-glucosidase(NAG enzyme)There were no significant differences between the three groups,urine ?2-microglobulin(?2-MG),and urine ?1-microglobulin(?1-MG)(P>0.05).3.Changes in serum biochemical indicatorsAfter 4 weeks of modeling,except for the blank control group,the SCr and BUN of the other groups were not significantly different from the model control group(P>0.05).After 8 weeks of administration,the serum cystic inhibitory level of the model control group rats C(CysC),BUN and SCr were significantly higher than those in the blank control group(P<0.01);Yishen Fang's low,medium,and high doses,and the uremic clearance positive control group had the above indexes lower than the model control group(P<0.05),The above indexes of Yishenfang high-dose group are closer to the standard(P<0.01);there is no significant difference in blood calcium(Ca)and blood phosphorus(P)in each group(P>0.05).4.Renal histopathological changesGross anatomy shows that the size,color,and morphology of the kidneys of rats in the blank control group were normal,and the boundaries of the cortex and medulla wereobvious.The kidneys of the other groups of rats increased in size and lightened in color.The appearance of the model control group,s rats changed most significantly,showing“large white kidneys”,and some rats in the model control group showed hydronephrosis and congestion in the kidney tissue.Cortical medullary boundaries are blurred.In each administration group,the color of the kidneys became red,the volume became smaller,and the boundaries of the cortex and medulla were still clear,which were almost normal.HE staining: In the blank control group,the glomerulus,tubule structure and morphology were normal,no inflammatory cell infiltration and no fibroblast proliferation.Compared with the blank control group,the number of glomeruli in the model control group decreased,the quality of the tubules decreased,the tubules were compensated to expand,and a large amount of xanthine metabolites were deposited in the tubes.The invasion of proteinuria,fibroblasts,and a large number of inflammatory cells was visible.Compared with the model control group,the pathological changes of renal tissues in each administration group were significantly reduced,and the high-dose group of Yishenfang was the most obvious.The degree of pathological changes was in the high-dose group>medium-dose group>Uduqing positive control group>low-dose group.Masson staining: There was no significant change in glomeruli in the blank control group,no significant metabolite deposition in the lumen of the renal tubules,and only a small amount of blue collagen fibers were deposited in a strand shape in the renal interstitial;compared with the blank control group,the model control The glomerular balloon of the rats in the group was significantly dilated,the basement membrane was significantly thickened,and large pieces of blue collagen fibers were deposited in the tubulointerstitial and epithelial cells.Compared with the model control group,the degree of fibrosis in each group was significantly reduced.Yishenfang high-dose group was the most obvious,and the degree of fibrosis reduction was in the order of high-dose group>medium-dose group>Uduqing positive control group>low-dose group.5.Changes in serum FGF23 expressionThe model control group was significantly higher than the blank control(P<0.01);the administration groups were significantly lower than the model control group(P<0.05),and the expression of FGF23 in the Yishenfang high-dose group was significantly reduced(P<0.01).6.Changes in Klotho mRNA expression in kidney tissueThe model control group was significantly lower than the blank control group(P<0.01);the administration groups were significantly higher than the model control group,the expression of Klotho mRNA in the Yishenfang high-dose group was significantly increased(P<0.05).7.Changes of TGF-?1 expression in kidney tissueThe model control group was significantly higher than the blank control group(P<0.01);the high-dose and uremic-clear positive control group of Yishenfang was lower than the model control group(P<0.05);There was no significant difference in the groups(P>0.05).Conclusion:1.Yishenfang can significantly improve the general condition,biochemical indicators and pathological changes of rats with early adenine-induced chronic renal failure,and it can obviously protect the kidney structure and function of rats with early chronic renal failure.2.Renal interstitial fibrosis in rats with early adenine-induced chronic renal failure may be closely related to the low expression of Klotho mRNA in kidney tissues and the high expression of serum FGF23 and TGF-?1 factor in kidney tissues.The expression of mRNA down-regulates the expression of serum FGF23 and TGF-?1 factor in kidney tissues,thereby generating renal protection. |
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