| ObjectiveTo investigate the effects of eighteen kinds of nephrotoxic Chinese drugs on three major types of Organic Anion Transporters, Oat1,Oat2,Oat3and several drug metabolizing enzymes in vivo in mice.MethodsWater decoction of eighteen kinds of nephrotoxic Chinese drugs was orally administered to NIH mice (50%of each gender per group) respectively at high and low dosages two times a day for5consecutive days. Also, there were groups of pure water control, Probenecid (positive control), Carboxymethylcellulose sodium (CMC, vehicle for Probenecid), Rifampin, Ketoconazole. In experimental day, one hour after the last dose at the5th day, all the mice were intravenously given para-aminohippuric acid (PAH) at a dose of30mg/kg. Mice (n=10/group) were euthanized and whole blood were collected at1,2.5,5,7.5,10,15,20min respectively. Right kidneys were collected immediately after bleeding. Another experimental day, NIH mice (10mice/group, both sex50%each) were treated with investigated samples and were euthanized after60min, kidneys were removed quickly. Another experimental day, NIH mice (10mice/group, both sex50%each) were killed after60min of treated with investigated samples, the liver, intestine and kidneys were removed quickly. Right kidney of PAH clearance test was homogenized for PAH accumulation test, part of right kidneys were for PAH uptake test for kidney slices, liver and intestinal microsomes were prepared by differential centrifugation, kidney homogenate and microsomal protein were quantified by Folin-phenol method, The concentrations of PAH in serum and kidney homogenates, as well as the activity of Cyp3a and Cyp2e1, were determined by spectrophotometry. Pharmacokinetic analysis of PAH in blood was done by pharmacokinetic Software (DAS2.0). The other part of left kidneys for extracting total mRNA in CYPs test were prepared to analysis of Oat1,Oat2and Oat3gene expressions using quantitative real-time PCR.ResultsCompared with water control t1/2β of high dosage of herba Leonuri, sophora flavescens, corydalis yanhusuo, angelica pubescens, cacumen biotae, both high and low dosage of scolopendra, kusnezoff monkshood and polygonum cuspidatum were significantly prolonged (P<0.05～0.01); Vd of all groups were significantly decreased (P<.0.01); except of low dosage of asarum, herba leonuri, angelica pubescens, spina gleditsiae, kansui root, CL and AUC0-20min of all other groups were considerably changed, CL reduced (P<0.01) and AUCo-20min increased (P<0.01). Except of low dosage of cordyceps sinensis, Scolopendra, pulsatilla, kusnezoff monkshood, PAH accumulation in kidneys in all other groups was extensively increase (P<0.05～P<0.01). Renal PAH uptake results in high dosage of corydalis yanhusuo, kansui root, costusroot, both high and low dosage of kusnezoff monkshood, spina gleditsiae and cacumen biotae were significantly lower (P<0.05～0.01) than corresponding control group. Oat1, Oat2,Oat3mRNA expressions in eight kinds of samples tested showed that all the corresponding genes of above transporters were obviously regulated.Liver Cyp3a activites, erythromycin as the substrate, both high and low dosage of asarum, scolopendra, sophora flavescens, angelica pubescens, high dosage of senna, polygonum cuspidatum, costusroot were significantly higher than control group (P<0.05); both high and low dosage of cinnamon, cordyceps sinensis, herba leonuri, magnolia officinalis, kusnezoff monkshood, monkshood, kansui root, cacumen biotae, low dosage of spina gleditsiae, costusroot were significantly lower than control group (P<0.05-0.01).Liver Cyp3a activites, aminopyrine as the substrate, both high and low dosage of asarum, scolopendra, sophora flavescens, corydalis yanhusuo, angelica pubescens, polygonum cuspidatum, high dosage of senna, spina gleditsiae, costusroot were significantly higher than control group (P<0.01); both high and low dosage of cordyceps sinensis, herba leonuri, magnolia officinalis, monkshood, high dosage of protoanemonin, kusnezoff monkshood, low dosage of spina gleditsiae, costusroot were significantly lower than control group (P<0.05～0.01).Cyp3a activities in intestines of high dosage of asarum, scolopendra and angelica pubescens, both high and low dosage of polygonum cuspidatum and costusroot, erythromycin and aminopyrine as the substrate, were significantly higher than control group (P<0.05～0.01); high dosage of cinnamon, kansui root, low dosage of protoanemonin, cordyceps sinensis, sophora flavescens, magnolia officinalis, corydalis yanhusuo, monkshood, cacumen biotae were significantly lower than control group (P<0.05-0.01).Hepatic Cyp2el activites in both high and low dosage of cinnamon, herba leonuri, scolopendra were obviously lower than control group (P<0.01); high dosage of cordyceps sinensis, senna, magnolia officinalis, spina gleditsiae, kansui root, both high and low dosage of asarum, sophora flavescens, protoanemonin, corydalis yanhusuo, kusnezoff monkshood, monkshood, angelica pubescens, cacumen biotae, costusroot was dramatically higher than control group (P<0.01).ConclusionEighteen renal toxic traditional Chinese drugs have a significant role in18kinds of traditional Chinese medicine for renal toxicity in mouse liver and intestine and liver Cyp3a Cyp2e (inhibition or induction). Eighteen renal toxic traditional Chinese drugs probably induced kidney damage through inhibiting several members of the Organic Anion Transporters (such as OAT1, OAT2, OAT3). |
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