| Background:the prevalence of chronic renal failure(CRF)is increasing year by year.It has the characteristics of high prevalence,low awareness,high medical cost,poor prognosis and high mortality.Modern medicine has no ideal treatment for CRF to delay the progress of renal function.Recent studies show that Klotho protein as an anti-aging factor and a renal protective factor,has a role in delaying renal aging,and is expected to become a predictive indicator and therapeutic target of CRF.In recent years,Chinese medicine widely used in patients with CRF,especially in the early and middle patients,according to the protection of residual renal units,improve symptoms and delaying the start of renal replacement therapy time and improve the quality of life and prolong the life of the principle,delay the progression of renal function,showing good advantages and characteristics.Shen Ning II,as a empirical prescription of Professor Yao Yuanzhang’s treatment of CRF,has achieved an ideal effect in the early clinical observation.Shen Ning II is mainly composed of Radix Astragali,Radix Polygoni Multiflori,Radix et Rhizoma Rhei,Ligusticum chuanxiong,leech,peach kernel and Eupatorium.It has the functions of tonifying kidney,purging turbid and promoting blood circulation to dissipate blood stasis,and has studied the protective mechanism of peach kernel,zehira and leech on the kidney.However,the protective mechanism of Astragalus membranaceus and Radix Polygoni Multiflori on the kidney has not yet been elucidated,and whether it is related to the protective factor Klotho of kidney is still needed to be further explored.Objective:To observe the rat model of CRF and traditional Chinese medicine intervention group model of correlation between the changes of renal function,calcium phosphorus metabolism and Klotho protein;By using Radix Astragali,Radix Polygoni Multiflori,Astragalus Radix Polygoni multiflorum mixture interfere with the rat model of CRF,to observe the effect of each group on the Klotho proteins in the kidney tissue,preliminary study of kidney tonifying traditional Chinese medicine Astragalus,Radix Polygoni Multiflori on kidney the protective mechanism.Methods:CRF rat model was established by adenine gavage.After the end of modeling,blood urea nitrogen(BUN),serum creatinine(Scr),blood phosphorus and blood calcium were measured.24h urine was collected,urine volume was measured,urine creatinine(Ucr),urine phosphorus and urinary calcium were measured,and screened according to the results.After modeling,they were randomly divided into three groups:Astragalus high group(HG),Astragalus low group(HG),Shouwu high group(SG),Shouwu low group(SD),huangshougao mixture high group(HSG),huangshou mixture low group(HSD).The drug intervention group in each group was given the corresponding concentration of drugs for the treatment of the stomach.The model group and the normal group were given the same amount of normal saline for gastric perfusion.During the period of drug intervention,the body weight and the general changes of observation records were observed every week.After the end of drug intervention determination of blood and urine biochemical indexes,Western blotting(Western Blot)to detect the expression of Klotho protein in renal cortex,observed by hematoxylin and eosin staining under light microscope(HE)change of renal pathology,positive immunohistochemical detection of Klotho protein in renal tubule staining.SPSS21.0 software was used for statistical comparison.Results:(1)weight changes:after the intervention,the weight of the model group did not increase significantly(P>0.05),but the weight gain of the other drug intervention groups was not significantly different from that of the normal group(P<0.01).(2)renal function changes:there was no significant difference between the normal group and the model group before and after intervention(P>0.05).The serum creatinine and urea nitrogen in each administration group were improved,and the creatinine excretion increased,but the renal function improvement in HG(P<0.01)was better than that in the other administration groups.(3)changes of endogenous creatinine clearance rate:high and low Huangqi g,Shouwu high and low group,the first group of yellow mixture of low renal function improved significantly(P<0.01),high group renal function than other yellow first mixture treatment group were significantly improved(P<0.05),normal group,model group showed no significant meaning.(4)the change of calcium and phosphorus metabolism:normal group and model group did not change significantly(P>0.05),and the administration of each group had different effects on regulating calcium and phosphorus metabolism(P<0.05).The high and low group,Huangqi group to promote the first yellow mixture of low urinary phosphorus excretion,reduce blood phosphorus effect;high and low group,Huangqi group,Shouwu high yellow first group promoting agent for high urinary calcium excretion was significantly higher and lower,Huangqi group,Shouwu high and low group of calcium significantly decreased(P<0.01),poor efficacy of yellow first mixture.(5)pathological changes of kidney:the yellow brown crystals of different sizes were found in the renal tubules and renal interstitium in each group,and a few were seen in the glomerulus.Dividing medullary renal cortex of rats in model group is not clear,and the cortex was thinner,glomerular atrophy,balloon lumen becomes large,cystic wall thickening,lobulated capillary plexus obviously,vascular loop opening is poor,I see glomerular artery thrombosis;renal tubular epithelial cell degeneration,necrosis and shedding,lumen expansion,showed a large amount of protein tube,renal interstitial fibrous tissue hyperplasia,accompanied by inflammatory cell infiltration.Huangqi high group glomerular structure change slightly,no obvious glomerulosclerosis atrophy,lobulation,small renal tubular epithelial cell degeneration and necrosis,cavity expansion,visible focal renal interstitial fibrosis,with a few scattered inflammatory cell infiltration.Huangqi low group mesangial matrix of moderate hyperplasia,mild differential capillary plexus leaflike,balloon lumen;renal tubular lumen dilated,with a small amount of epithelial cell degeneration and necrosis,patients with high Huangqi was more serious.Glomerular sclerosis and enlargement of the cystic cavity were obvious in Shouwu high group.The capillary loops were poorly opened,tubule necrosis and shedding were obvious,interstitial fibrosis and inflammatory cell infiltration were significant,but the lesions were slightly more than those in the model group.The glomerulonephritis in Shouwu low group was characterized by moderate glomerulonephritis,a large number of renal tubular necrosis and abscission,interstitial fibrosis and infiltration of inflammatory cells,with a small number of protein tube types,but the lesions were lighter than those in Shouwu high group.The first high yellow mixture group focal segmental glomerulosclerosis,cavity expansion,proliferation of mesangial matrix and basement membrane thickening,vascular plexus lobular,open poor renal tubular degeneration,necrosis and lumen increased,patients,interstitial fibrosis and inflammatory cell infiltration.The first yellow mixture of low group glomerulus sclerosis atrophy,no significant vascular loop opened well,cavity expansion,wall thickening,tubular degeneration and necrosis,bureaucratic change,renal interstitial fibrosis and inflammatory cell infiltration in focal lesions,is the first high yellow mixture group compared with ease.(6)immunohistochemistry:the expression of renal tissue were detected Klotho protein,the normal group,model group at each dose group compared with model group,Klotho protein expression increased(P<0.05),in which the high and low group,Huangqi group,Shouwu low yellow first mixture of low group more significant effect(P<0.01).(7)Western Blot results:the expression of Klotho protein was detected in all renal tissues,and the expression of Klotho protein in the normal group was the highest,and the expression of the model group was least.The expression of Klotho protein increased in each drug delivery group(P<0.05),and the expression of low histone in high and low Astragalus group,Shouwu high and low group and huangshou mixture increased more significantly(P<0.01).(8)correlation with the biochemical index of Klotho protein:with the increased expression of renal Klotho protein in rats,Ccr,urinary phosphorus,urinary calcium increased,blood urea nitrogen,phosphorus decreased significantly,renal function improved significantly,suggesting that the Klotho protein and Ccr,urine phosphorus,urinary calcium were positively correlated,negatively correlated.Relationship between serum creatinine and urea nitrogen.However,the increase of Klotho protein showed no significant decrease in blood calcium.Conclusion:Bushen Chinese Medicine Radix Astragali,Radix Polygoni Multiflori can up regulate expression of Klotho in kidney of protein,play the renal protection by reducing serum creatinine,urea nitrogen,serum phosphorus,increased creatinine clearance and urine phosphorus,urinary calcium excretion,improve renal function and related symptoms,reduce the damage of renal tissue pathological structure.Delay the progression of renal function,the high dose group had the best effect of huangqi. |
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