| ObjectiveIn recent years,the incidence of chronic kidney disease(CKD)has gradually increased.However,patients usually are unaware of CKD until middle and terminal stages due to the limited change of biochemical index and early clinical symptoms.The late treatment leads to huge burden of the family and society.In order to reduce the dangers of CKD,it is greatful for studying its early diagnosis and treatment.Previous studies comprehensively analyzed urine protein from primary disease of chronic glomerulonephritis patients with CKD by urinary proteomics techniques.Results showed that,along with the progress of the disease,CPN1 protein in the urine of CKD patients changed obviously.However,there is a lack of related studies on whether CPN1 can be a biomarker for the CKD progression.CKD belongs to traditional Chinese medicine etiology "retention of urine","block and repulsion","edema".Traditional Chinese medicine theory for CKD including spleen and kidney deficiency,turbid poison and blood stasis resistance,the real virtual pathogenesis.Based on this theory,famous traditional Chinese medicine professor Qinguo Hong from Guangdong province developed Shengqingjiangzhuo capsule,which has been successfully applied to clinical treatment for more than 20 years.Shengqingjiangzhuo capsule is composed of traditional Chinese medicine such as,Huangqi,Dahuang,Danshen,Faxia,Zhike,Tuful ing.Previous cl inical studies and animal studies showed that,Shengqingjiangzhuo capsule has the effect of renal protection and delay the progression of CKD.By comparing the urine protein spectrum of chronic glomerulonephritis patients with CKD before and after Shengqingjiangzhuo capsule treatment,results showed that Shengqingjiangzhuo capsule can affect the level of urine CPN1 significantly.However,the mechanism of CPN1 in the progression of CKD has not been elucidated.Therefore,the purpose of this study includes the following:(1)To reveal the changes of CPN1 in urine and kidney of 5/6 nephrectomy rats and the role of CPN1 in CKD progression.(2)To clarify the relationship between Shengqingjiangzhuo capsule and CPN1 in 5/6 nephrectomy rats and the mechanism of Shengqingj iangzhuo capsule on CKD.Methods(1)Establishment of CKD modelThis experiment adopted 5/6 nephrectomy rat model,and were divided into six groups:normal group,sham operation group,5/6 nephrectomy group,Shengqingjiangzhuo capsule treatment group including low dosage group,medium group and high dosage group;(2)Collection of samples1 week,2 weeks,4 weeks,8 weeks after 5/6 nephrectomy,samples including blood,urine,kidney tissues,etc were collected;(3)Biochemical IndicatorsRenal function determined by automatic biochemical analyzer,urine analysis determined by urine analyzer;renal pathology examination:HE staining,Masson staining and immunohistochemical;CPN1 mRNA expression in kidney determined by RT-PCR;CPN1 protein expression in urine and kidney determined by the Mouse CPN1 ELISA kit.Results(1)The effect of Shengqingjiangzhuo capsule treatment on 5/6 nephrectomy rat renal function and urine protein8 weeks after Shengqingjiangzhuo capsule treatment,serum creatinine,serum urea nitrogen and 24 h urine protein were significantly decreased in low dosage group(800 mg/kg),medium dosage group(1200 mg/kg),while high dosage group(1600 mg/kg)showed significant effect on urea nitrogen.From the above three indices synthetically,the medium dosage treatment showed the best curative effect,followed by low dosage group and high dose group.(2)The effect of Shengqingjiangzhuo capsule treatment on 5/6 nephrectomy rat kidney pathological morphology8 weeks after Shengqingj iangzhuo capsule treatment,renal pathological change of medium dosage(1200 mg/kg)was minimized.Hyperplasia in glomerular basement membrane and mesangium,atrophy and necrosis in minority of renal tubular,infiltration in interstitial inflammatory cell and hyperplasia in fibrous connective tissue were also observed in the two groups.In high dosage group(1600 mg/kg),hyperplasia in glomerular basement membrane and mesangium was obviously observed,necrosis in renal tubular was relatively serious,numerous inflammatory cell infi ltration in renal interstitial and hyperplasia in fibrous connective tissue were also observed.Therefore,the medium dosage treatment showed the best curative effect on renal fibrosis of rat with CKD.(3)Shengqingjiangzhuo capsule treatment effect on CPN1 in 5/6 nephrectomy rat kidneymRNA level:8 weeks after Shengqingjiangzhuo capsule treatment,CPN1 mRNA expression was significantly increased,and the r i sing trend was:low dosage group(800 mg/kg)<high dosage group(1600 mg/kg)<medium dosage group(1200 mg/kg).Therefore,the medium dosage treatment showed the best effect on CPN1 mRNA expression.Protein detection:Shengqingj iangzhuo capsule treatment groups showed higher CPN1 level in the urine over the 5/6 nephrectomy group,and higher dosage showed more obvious r ising trend.However,there was no statistical difference,therefore,high dosage showed the best effect on CPN1 protein level in urine.Similar trend was showed in kidney.Shengqingj iangzhuo capsule treatment groups showed higher CPN1 level over the 5/6 nephrectomy group,and higher dosage showed more obvious r ising trend.However,there was no statistical difference.Therefore,high dosage showed the best effect on CPN1 protein level in kidney.ConclusionIn 5/6 nephrectomy group,CPN1 were clearly lower than the control group in both mRNA and protein levels,indicating that CPN1 can be used as a reference index in early diagnosis of CKD.8 weeks of medium dosage(1200 mg/kg)Shengqingjiangzhuo capsule treatment reduced serum creatinine,serum urea nitrogen levels,24 h urine protein,partly restore renal function;reduced in glomerular sclerosis and renal tubular interstitial fibrosis index.It slowed the renal fibrosis progression and improved CPN1 expression in both mRNA and protein level,indicating that Shengqingjiangzhuo capsule had kidney protection effect through the regulation of CPN1. |
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