| Objective:To investigate the effects on rat kidney caused by different dosage forms of Aristolochia debilis Sieb.et Zucc(RADSZ)and complex prescription contains RADSZ such as RADSZ pulveres,guanxinsuhe pill and RADSZ decoction,and explore the influence of the aristolochic acid(AA)A content of RADSZ caused by it's different dosage form.Methods:Healthy adult wistar rats(200±10g,♀:♂=1:1)were divided randomly into four groups(n=20 per group):RADSZ pulveres group, guanxinsuhe pill group,RADSZ decoction group and the control group. The dosage is RADSZ pulveres 0.81g·kg-1·d-1,guanxinsuhe pill 0.32g·kg-1·d-1,RADSZ decoction 0.81g·kg-1·d-1,the drugs were intragastric administration for rats respectively for three continuous months.While the control group were given distilled water in the same volume.We have weighing each rat's bodyweight every week,and the dosage was changed following it's bodyweight.We have examined the level of SCr and BUN in blood,the level ofβ2-MG and NAG in urine every month.At the end of the third month,all the rats were sacrificed and the renal pathological examination of each rat was performed.The protein expression of TGF-β1 and CTGF was determined by immunohistochemistry and the mRNA expression of CTGF was determined by real time PCR method,respectively. The AAA in nephridial tissue samples,RADSZ pulveres,guanxinsuhe pill and RADSZ decoction were determined by HPLC Method.Results:(1)In RADSZ decoction group,the level of SCr and BUN in blood, the level ofβ2-MG and NAG in urine at the end of frist,second and third month showed no statistical differences compared with the control group(P＞0.05).Histological examination did not reveal significant abnormality in the kidney at the end of third month.Immunohistochemistry showed that protine expression of TGF-β1 and CTGF in nephridial tissue were slight up-regulated,and was no statistically significant compared with the control group(p＞0.05).Real time PCR showed that the expression of the CTGF mRNA were respectively the 1.14 times compared with the control group.HPLC method showed that the content of AAA in nephridial tissue samples were respectively 0.1045μg/g.(2)In RADSZ pulveres group, the level of SCr and BUN in blood at the end of first,second and third month showed no statistical differences compared with the control group (P＞0.05),but the level ofβ2-MG and NAG in urine increased at the end of second and third month compared with the control group(p＜0.05 or P＜0.01),and was statistically significant compared with the first month (p＜0.05),and the level ofβ2-MG and NAG in urine increased at the end of third month compared with the second month(p＜0.05). Immunohistochemistry showed that protine expression of TGF-β1 and CTGF in rat kidney tissue were significant up-regulated,and was statistically significant compared with the control group(p＜0.01).Real time PCR showed that the expression of the CTGF mRNA were respectively the 2.83 times compared with the control group.The renal pathologic examination showed tubular-interstitial injury.HPLC method showed that the content of AAA in nephridial tissue samples were respectively 0.8555μg/g.(3)In guanxinsuhe pill group,the level of SCr and BUN in blood at the end of first,second and third month showed no statistical differences compared with the control group(P＞0.05),but the level ofβ2-MG and NAG in urine significantly increased at the end of second and third month compared with the control group(p＜0.05 or P＜0.01),the level of NAG andβ2-MG in urine taken at the end of the third month obviously increased,and was statistically significant compared with the first month and the second month(p＜0.05 or P＜0.01). Immunohistochemistry showed that protine expression of TGF-β1 and CTGF in rat kidney tissue were significant up-regulated,and was statistically significant compared with the control group(p＜0.01).Real time PCR showed that the expression of the CTGF mRNA were respectively the 2.21 times compared with the control group.The renal pathologic examination showed tubular-interstitial injury.HPLC method showed that the content of AAA in nephridial tissue samples were respectively 0.2173μg/g.(4)HPLC method showed that the content of AAA in RADSZ pulveres,guanxinsuhe pill and RADSZ decoction were 1.082 mg/g,0.1542 mg/g,0.1342 mg/g.Conclusion:1.The intragastric administration with RADSZ pulveres, guanxinsuhe pill which lasted three continuous months can cause different levels injury of the renal tubule-interstitial substance.2.The intragastric administration of RADSZ decoction for rat for three continuous months cann't cause significant injury of the renal tubule-interstitial substance.3. Differert types of the RADSZ has a huge diversty in content of aristolochic acid,the dissovled amour of AAA in decoction dosage form was lower than in solid dosage form.
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