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OBJECTIVE: To discuss the mechanism of Yiqiyangyin recipe in rats with adriamycin (ADR)-induced nephropathy. METHODS: We randomly divided 30 Sprague-Dawley rats into 5 groups: control, model, glucocorticoid, Chinese herb, and Chinese herb plus glucocorticoid 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 serum 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 immunosorbent assay. The expression of nephrin was detected by immunohistochemical staining and quantitatively analyzed using Motic image analysis 3.2 software. RESULTS: The levels of urinary protein and serum triglycerides and cholesterol in rats with ADR-induced nephropathy increased on day 14. The serum albumin levels simultaneously decreased. All the changes reached the peak on day 28. Examination under the light microscope showed inflammatory cells and slight fibroplasia in the renal interstitium 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 expression was higher in the model group than that in the control group. The HPA mRNA levels in the intervention 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 significantly with the proteinuria level. No significant difference was found in the HPA level in urine between the intervention groups and the model group, whereas the model group had a higher urinary 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 glucocorticoid group. Compared with the control group, themodel group showed increased nephrin expression in the kidney. Nephrin levels in the other groups, especially 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 regulate the expression of nephrin in rats with ADR-induced nephropathy. Our study showed that treatment with Yiqiyangyin recipe plus glucocorticoid was better than a singular intervention, and we explored the pharmacological mechanism of this combination by biochemical and molecular biological analysis to provide a basis for clinical application.
OBJECTIVE: To discuss the mechanism of Yiqiyangyin recipe in rats with adriamycin (ADR) -induced nephropathy. METHODS: We randomly divided 30 Sprague-Dawley rats into 5 groups: control, model, glucocorticoid, Chinese herb, and Chinese herb plus glucocorticoid 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 serum 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 immunosorbent assay. The expression of nephrin was detected by immunohistochemical staining and quantitatively analyzed using Motic image analysis 3.2 software. RESULTS: The levels of urinary protein and serum triglycerides and cholesterol in rats with ADR-induced nephropathy increased on day 14. All the changes reached the peak on day 28. Examination under the light microscope showed inflammatory cells and slight fibroplasia in the renal interstitium 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 expression was higher in the model group than that in the control group. The HPA mRNA levels in the intervention 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 significantly with the proteinuria level No significant difference was found in the HPA level in urine between the intervention groupsand the model group, whereas the model group had a higher urinary 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 glucocorticoid group. compared with the control group, the knockout group showed increased nephrin expression in the kidney. Nephrin levels in the other groups, especially in the Chinese herb plus glucocorticoid group, were significantly lower than that that in the model group. The nephrin levels were the 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 regulate the expression of nephrin in rats with ADR-induced nephropathy. Our study showed that treatment with Yiqiyangyin recipe plus glucocorticoid was better than a singular intervention, and we explored the pharmacological mechanism of this combination by biochemical and molecular biological analysis to provide a basis for clinical application.