Queensland Cents Capsule Pharmacodynamic Study Of Rats With Adriamycin Nephrosis

Nephrotic syndrome is a glomerular disease caused by various pathogens,characterized by proteinuira＞3.5g/24h,hypoproteinemia(plasma albumin＜30g/L),edema and hyperlipidemia.The current therapy is focused on glucocorticoid and cytotoxic agents which are not specificly selective,resulting in toxic and side effects.Therefore,exploration of Chinese medicine with ascertained therapeutic effect and less toxic and side effects is critical for the treatment of nephrotic syndrome.Rat model with adriamycin-induced nephropathy was initially reported by BertaniⅠin the early 1980s and is a widely accepted model with similar pathological lesions with human glomerular disease.The glomerular lesions manifest as minimal change glomerular disease and focal segmental glomerulosclerosis,with clinical manifestion of nephrotic syndrome.Rat model with adriamycin-induced nephropathy can be classified as two types:acute and chronic types.The acute type is induced by single caudal vein injection of adriamycin at dosage of 5mg/kg-7.5mg/kg,with glomerular lesions similar with human minimal change disease.The chronic type is characerized by focal segmental glomerulosclerosis which is indjuced by several caudal injection of adriamycin or in combination with single nephrectomy.Adriamycin model is simple to establish,with consistent lesions and little inter-individual deviations.Kunxian capsule is produced by pharmaceutical factory of Chen Liji in Guangzhou, which is effective in immune inhibition and antagonism of cytokines,antiinflammtion and analgesia,dispelling the wind to remove the pain and reinforcing kindey to communicate the meridians and channels.It is clinically applied to treat autoimmune diseases such as rheumatoid arthritis and ankylosing spondylitis and found to be effective in decreasing albuminuria level.In order to prove this effect and provide laboratory evidence for clinical application,this study is undertaken to observe the therapeutic effect of Kunxian capsule on adriamycin-induced nephrotic rats.Objective:1.To establish an optimized rat model of adriamycin-induced nephrotic syndrome.2.To observe the therapeutic effect of Kunxian capsule on rat model of nephritic syndrome induced by adriamycin.Method:1.Establishment of adriamycin rat model:According to literature,A single caudal vein injection of adriamycin at doses of 5 mg/kg,6 mg/kg,7 mg/kg and twice caudal injection of adriamycin with a total dose of 7.5 mg/kg are screened to optimize model establishment. Twenty-four hours proteinuria were measured to evaluate the change curve and stability of the model,blood biochemical parameters and pathological changes are compared between the model and control groups.2.Seventy male Wistar rats were randomly divided into six groups:the normal control group, the model group,the glucocorticoid group,the Kunxian capsule group of high dose,the Kunxian capsule group of median dose and the Kunxian capsule group of low dose.Rat model of nephritic syndrome was established by caudal vein injection of adriamycin at a dose of 6 mg/kg,while the rats in the control group were injected the same volume of saline. Seven days before model establishment,rats in the glucocorticoid group were intragastricly administered prednisone at a dose of 7.5 mg/(kg.d);the Kunxian capsule groups of high, median and low group were intragastricly administered at doses which were equivalent with crude herbs of 30 g/(kg·d),15 g/(kg·d)and 7.5 g/(kg·d),equvilant with 30,15 and 7.5 times of human dose,suspended in 0.5%gum tragacanth solution.Intragastric administration was performed continually for 52 days.The effect of Kunxian capsule on body weight and 24h urinary protein was observed in the rat with kidney disease induced by adriamycin.the rats were sacrificed at the 53rd day of experiment.Serum total protein,albumin,total cholesterol, triglyceride,blood urea nitrogen and creatinine was tested.Kidney tissues were obtained for pathological examination and semi-quantitative score.Result:1.The group of twice injection of adriamycin exhibited more problems.Drug leakage observed after the first injection resulted in ulcerage of rat tails and failure of the second injection,thus increased inter-individual deviation in the rats.Proteinuria level in rats developed slowly at dose of 5 mg/kg which cannot be ascertained as nephrotic syndrome.Ptoteinuria level in rats with 7mg/kg injection differed dramaticly which increased the value of standard deviation.Individual difference was less significant and proteinuria level was relatively high and stable with caudal injection of adraimycin at a dose of 6 mg/kg, with proteinuria level increased one week after adriamycin injection and presented typical pathological changes of nephrotic syndrome at the 6th week,which was high level of proteinuria,hypoalbumia, hyperlipidemia.Median hypertrophy of mesangial cells,tubular atrophy and interstitial inflammatory cell infiltration were observed under the microscope.2.Compared with the normal control group,24h urinary protein and blood lipid levels of the model group were significantly elevated(P＜0.01)and the serum albumin level was obviously decreased(P＜0.01).Compared to those in the model group,24h urinary protein levels in the glucocorticoid and Kunxian groups were significantly decreased(P＜0.01,P＜0.05 respectively);blood lipid levels were significantly decreased(P＜0.01);in the glucocorticoid group and Kunxian groups of low and median dose group,the glomerular injury was ameliorated(P＜0.01);in the glucocorticoid group and Kunxian groups of high and median dose group,the tubulointerstitial injury was ameliorated significantly(P＜0.01);Conclusion:1.Rat model established by one single caudal injection of adriamycin at a dose of 6 mg/kg exhibit stable increase of proteinuria level,with good to lerance of rats and no self-restriction.2.Rat model of nephritic syndrome can be successfully established by caudle vein injection of adriamycin at a dose of 6 mg/kg.Kunxian capsule is effective in the treatment of chronic kidney disease in rat induced by adriamycin and the median dose of Kunxian capsule showed the best effect.