OBJECTIVE： To discuss the mechanism of Yiqiyang- yin recipe in rats with adriamycin （ADR）-induced ne- phropathyo METHODS： We randomly divided 30 Sprague-Daw- ley rats into 5 groups： control, model, glucocorti- co...OBJECTIVE： To discuss the mechanism of Yiqiyang- yin recipe in rats with adriamycin （ADR）-induced ne- phropathyo METHODS： We randomly divided 30 Sprague-Daw- ley rats into 5 groups： control, model, glucocorti- coid, Chinese herb, and Chinese herb plus glucocor- ticoid groups. The 24-h urine volume was collected on days 7, 14, 21, and 28 after ADR injection, and all rats were killed on day 28. We measured the 24-h levels of urinary protein, serum cholesterol, and se- rum triglycerides, and renal function of all rats by using routine biochemical methods. Pathological changes in the rat kidneys were observed under light and electron microscopes. Heparanase （HPA） mRNA expression levels were measured using re-al-time fluorescence-quantitative polymerase chain reaction. Urine levels of HPA in all groups were measured using enzyme-linked immunosor- bent assay. The expression of nephrin was detected by immunohistochemical staining and quantitative- ly analyzed using Motic image analysis 3.2 software. RESULTS： The levels of urinary protein and serum triglycerides and cholesterol in rats with ADR-in- duced nephropathy increased on day 14. The se- rum albumin levels simultaneously decreased. All the changes reached the peak on day 28. Examina- tion under the light microscope showed inflamma- tory cells and slight fibroplasia in the renal intersti- tium in the model group, but fewer inflammatory cells were observed in the intervention groups than those in the model group. Examination under the electron microscope showed extensive fusion of foot processes in ADR rats. HPA mRNA expres- sion was higher in the model group than that in the control group. The HPA mRNA levels in the in- tervention groups, especially in the Chinese herb group, and Chinese herb plus glucocorticoid group were significantly lower than the level in the model group. The HPA expression levels correlated signifi- cantly with the proteinuria level. No significant dif- ference was found in the HPA level in urine be- tween the intervention groups and the model group, whereas the model group had a higher uri- nary HPA level than the control group. Nephrin mRNA expression levels in the model group were higher than those in the control group. Nephrin mRNA expression levels were significantly lower in the intervention groups than that in the model group, especially the Chinese herb plus glucocorti- cold group. Compared with the control group, themodel group showed increased nephrin expression in the kidney. Nephrin levels in the other groups, es- pecially in the Chinese herb plus glucocorticoid group, were significantly lower than that in the model group. The nephrin levels in the kidney were negatively correlated with the proteinuria level. CONCLUSION： Yiqiyangyin recipe could attenuate foot process injury particularly in combination with a steroid reduce the development of proteinuria possibly by inhibition of HPA in the kidney, and reg- ulate the expression of nephrin in rats with ADR-in- duced nephropathy. Our study showed that treat- ment with Yiqiyangyin recipe plus glucocorticoid was better than a singular intervention, and we ex- plored the pharmacological mechanism of this combination by biochemical and molecular biologi- cal analysis to provide a basis for clinical applica- tion.展开更多
基金Supported by Shanghai Health Bureau Research Grant Program(Young Funding Program,No.2011QL031B)State Key Clinical Department of TCM pediatrics
文摘OBJECTIVE： To discuss the mechanism of Yiqiyang- yin recipe in rats with adriamycin （ADR）-induced ne- phropathyo METHODS： We randomly divided 30 Sprague-Daw- ley rats into 5 groups： control, model, glucocorti- coid, Chinese herb, and Chinese herb plus glucocor- ticoid groups. The 24-h urine volume was collected on days 7, 14, 21, and 28 after ADR injection, and all rats were killed on day 28. We measured the 24-h levels of urinary protein, serum cholesterol, and se- rum triglycerides, and renal function of all rats by using routine biochemical methods. Pathological changes in the rat kidneys were observed under light and electron microscopes. Heparanase （HPA） mRNA expression levels were measured using re-al-time fluorescence-quantitative polymerase chain reaction. Urine levels of HPA in all groups were measured using enzyme-linked immunosor- bent assay. The expression of nephrin was detected by immunohistochemical staining and quantitative- ly analyzed using Motic image analysis 3.2 software. RESULTS： The levels of urinary protein and serum triglycerides and cholesterol in rats with ADR-in- duced nephropathy increased on day 14. The se- rum albumin levels simultaneously decreased. All the changes reached the peak on day 28. Examina- tion under the light microscope showed inflamma- tory cells and slight fibroplasia in the renal intersti- tium in the model group, but fewer inflammatory cells were observed in the intervention groups than those in the model group. Examination under the electron microscope showed extensive fusion of foot processes in ADR rats. HPA mRNA expres- sion was higher in the model group than that in the control group. The HPA mRNA levels in the in- tervention groups, especially in the Chinese herb group, and Chinese herb plus glucocorticoid group were significantly lower than the level in the model group. The HPA expression levels correlated signifi- cantly with the proteinuria level. No significant dif- ference was found in the HPA level in urine be- tween the intervention groups and the model group, whereas the model group had a higher uri- nary HPA level than the control group. Nephrin mRNA expression levels in the model group were higher than those in the control group. Nephrin mRNA expression levels were significantly lower in the intervention groups than that in the model group, especially the Chinese herb plus glucocorti- cold group. Compared with the control group, themodel group showed increased nephrin expression in the kidney. Nephrin levels in the other groups, es- pecially in the Chinese herb plus glucocorticoid group, were significantly lower than that in the model group. The nephrin levels in the kidney were negatively correlated with the proteinuria level. CONCLUSION： Yiqiyangyin recipe could attenuate foot process injury particularly in combination with a steroid reduce the development of proteinuria possibly by inhibition of HPA in the kidney, and reg- ulate the expression of nephrin in rats with ADR-in- duced nephropathy. Our study showed that treat- ment with Yiqiyangyin recipe plus glucocorticoid was better than a singular intervention, and we ex- plored the pharmacological mechanism of this combination by biochemical and molecular biologi- cal analysis to provide a basis for clinical applica- tion.
