Yiqi Model About Heat Paste Puro Amino Nucleoside Nephropathy Rats Therapeutic Research

Objective:To establish a stable rat model with nephrosis induced by puromycin aminonucleoside (PAN) and then observe the renoprotective effect of replenish Qi and clear heat extract(益气清热膏)on PAN induced nephrosis.Method:1.15male Wistar rats that weighed100to110gram, were randomly divided into3groups: the control group with3rats, the PAN small dose group (40mg/kg weight)with6rats and the PAN large dose group (55mg/kg weight) with6rats. Rats of the experimental group were performed with injection of PAN at the dose according to the group via the left jugular vein. While the control group were given an injection of0.9%normal saline at the same volume. Then rats of the experimental group were given vena caudalis injections with PAN at a dose of0.5mg/kg, at13th,16th and19th days after the jugular vein injections. And the control group were performed with0.9%normal saline at the same volume. Total proteinuria of24hours were measured at0th,4th,8th,12th,15th,19th,24thand28th after the first injection. All the rats were sacrificed at day30, and blood was collected via aortaventralis. The levels of serum total protein, albumin, creatinine, urea nitrogen, cholesterol and triglycerin levels were examined, and renal tissue pathological alterations of each rat were observed.2.30male Wistar rat of100±10gram, were randomely divided into6groups:the control group, the PAN model group (40mg/kg), small dose group, medium dose group, large dose group and the fosinopril group, each group with5rats. Rats of the experimental group were performed with injection of PAN at a dose of40mg/kg via the left jugular vein. While rats of the control group were given an injection of0.9%normal saline at the same volume. Then rats of the experimental group were given vena caudalis injections with PAN at a dose of0.5mg/kg, at13th,16th and19th days after the jugular vein injections. And rats of the control group were performed with0.9%normal saline at the same volume. Replenish Qi and clear heat extract was administered orally at the dose described above according to groups. Total proteinuria of24hours were measured at0th,5th,10th,15th,20th,25th and30th after the first injection. All the rats were sacrificed at day30, and blood was collected via aortaventralis. Kidneys were dissected, and renal index was measured. The levels of serum total protein, albumin, creatinine, urea nitrogen, cholesterol and triglycerin levels were examined. Renal tissue was stained with periodic acid-Schiff reagents for evaluating pathological alterations of each rat, and immunohistochemistry was performed.Results: 1. Rats in the control group did not show albuminuria. The PAN large dose group showed a much larger amount of albuminuria, but did not have a regular pattern to follow. The albuminuria of PAN small dose group began to increase at the4th day, and reached its peak at the8th day, and decreased rapidly after the12th day. After vena caudalis injections, albuminuria levels decreased slowly and retained in plateau periods. Compared with control group, as light microscopy showed, the histologic appearance of glomeruli from each PAN nephrosis group was much more server, according to mesangial cell increase and inflammatory infiltration in the renal tissue. Immunohistochemical staining for nephrin in the kidneys was dramatically decreased in PAN induced nephrosis groups.2. Replenish Qi and clear heat extract of small dose showed prominent enhance of pathological alteration and less decrease of kidney nephrin.Conclusion:A stable rat model with nephrosis induced by puromycin aminonucleoside (PAN) could be established via a single jugular vein injection of PAN at a dose of40mg/kg and three vena caudalis injections of PAN at a dose of0.5mg/kg. Replenish Qi and clear heat extract can reduce albuminuria of PAN induced nephrosis, which may possibly be taken effect by protect kidney nephrin form decreasing.