Effects Of Compound Shenyanpian And Prednisoneon In Juvenile Rats With Adriamycin-induced Nephropathy And Mechanism Study

Objective:Primary nephrotic syndrome(PNS) is one of the most common glomerular disease in pediatric, long-term use of glucocorticoid hormone(GC) treatment can cause complications such as metabolic disorders, Cushing syndrome, and about 23% of children with PNS are not sensitive to hormone therapy, which are type of steroid-resistant nephrotic syndrome(SRNS) and rising year by year. Compound Shenyanpian contains the Chinese native medicine ingredient such as salvia miltiorrhiza and astragalus membranaceus, clinical used in children with PNS showed good auxiliary therapeutic effect. This study was based on adriamycin nephrosis animal model and carried to observe combined effects of compound Shenyanpian joint prednisone on adriamycin nephrosis rats as well as their effects on the expression of ET-1, TGF-β1. Methods:40 4-weeks SD rats were randomized to select the normal control group(group A), the remaining 32 rats accepted one-off tail intravenous injection at 6.5 mg/kg of adriamycin and were randomly divided into four groups, namely the model control group(group B), compound shenyanpian group(group C), prednisone group(group D), the combined treatment group(group E), there were eight groups in each group. The control group accepted the same amount of saline tail intravenous injection. From 1st ~ 12 nd week, all the rats accepted daily morning lavage, group A and B group was given distilled water to fill the stomach, group C accepted water extract of 6.75 g/kg to fill the stomach, group D accepted prednisone suspension liquid 6 mg/kg gavage, group E was given prednisone 6 mg/kg + compound shenyanpian of 6.75 g/kg, the gavage volume is 1 ml/kg. 24 hour urine protein were tested at 1 day before modelling, end of 2, 4, 12, 8 weeks.At the end of 12 weeks, blood albumin, blood lipid and renal function and pathologic changes were examined. At 1 day before modelling, end of 12 weekends ELISA method used to detect serum ET-1,TGF-β1,TNF-α and IL-18. q-PCR method were used to detect ET-1, TGF-β1, transmission electron microscopy(SEM) were used to exam changes of kidney organization. SPSS13.0 statistical software was used for statistical analysis, which was statistically significant at P < 0.05. Results:(1) In group A the urine protein levels at five time point showed no significant change(F=5.03;P>0.05), urine protein at each time point was significantly higher than group A(F=23.59;P < 0.05). Urine protein level of C, D, E group at the end of 8 and 12 weeks decreased significantly than B group(F=15.66;P < 0.05), group E at the end of 12 weeks than the control group showed no significant difference(F=7.04;P > 0.05) but showed significant decline than the C, D group(F=19.61;P < 0.05).(2) Albumin and total cholesterol, triglycerides of group B, group C, group D compared with group A showed significant difference(F Alb=27.13; FChol=19.80;FTG=19.05;P<0.05). Albumin and total cholesterol, triglycerides of E group showed no significant differences than A group(F Alb=8.41;FChol=6.69;FTG=7.25;P >0.05).(3) At the end of 12 weeks group E compared with group B, C, D showd lighter pathological damage, renal interstitial hyperplasia and protein type tube was not seen in renal tubule.(4) TNF-α, IL-18,ET-1,TGF-β1 level of group B, group C, group D at the end of 12 weeks than before modelling and A group was significantly higher(FTNF-α=14.93; FIL-18=19.43; F ET-1=16.23; F TGF-β1=17.97; P < 0.05), group C, D, E group at the end of 12 weeks after treatment compared with group B were significantly improved(FTNF-α=18.32; F IL-18=17.68; F ET-1=17.29; F TGF-β1=20.14; P < 0.05). group C, D group at the end of 12 weeks compared with group E were significantly improved(FTNF-α=20.75; F IL-18=15.34; F ET-1=22.57; F TGF-β1=20.54; P<0.05).。(5) q-PCR showed m RNA expression of ET-1 in group B, group C, group D were significantly higher than group A(F =14.96; P < 0.05), group C, D and E group compared with group B reduced significantly((F=19.41; P < 0.05). group E compared with group A were no significant differences(F=7.23;P>0.05). TGF-β1 m RNA expression of group B were significantly higher than group A(P < 0.05), C, D, E group after treatment compared with group B were significantly improved(P < 0.05).(6) The electron microscope examination showed mesangial cell proliferation, epithelial cells free bubble formation in group B, mesangial cell damage and renal tubular protein particles were relieved in group C and group D. Epithelial cells, mesangial area were not seen obvious abnormity, renal tubular can be found in a small amount of protein particles in group E. Conclusion:(1) Compound shenyanpian combined prednisone can obviously decrease the albuminuria, improve kidney function and reduce blood lipids level, delay the progress of the renal pathological damage in adriamycin nephrosis rats and down regulation of ET-1, TGF-β1 m RNA expression plays important role in alleviating kidney reconstruction process.(2) Compound shenyanpian combined prednisone than compound shenyanpian or prednisone alone has better efficacy for treatment of adriamycin nephrosis rats, its mechanism may be related to two single medicine through the different targets to achieve synergy effect.