The Study Of Renal Toxicity About ShuangHuangLian Intravenous Injection Combined With Cefepime Intravenous Injection

ObjectiveShuangHuangLian powder injection(SHLI)that contains main active ingredients chlorogenic acid and baicalin is used for antiviral treatment.Cefepime is the fourth generation cephalosporin that belong to beta lactam compounds.The OAT-1 transporter,a transporter which excrete organic anions from basolateral to apical of renal proximal tubular cells,is located on the side of basolateral membrane of epithelial cells of renal tubules.The aim of study is to investigate pharmacokinetics interaction of cefepime and SHLI to exaime that whether combination cefepime with SHLI might increase renal toxicity and the effect of Oatl affter 7d of SHLI.Methods2.1 The bioavailability of cefepime were effected by SHLI and its main ingredients chlorogenic acid and baicalin.An Agilent 1100 series liquid chromatographic system was used for measuring plasma concentration of cefepime.The analytes of cefepime and internal standard were separated on a C18 column(Luna,250mm×4.6mm Phenomenex Technologies,USA)and were detected under the UV=254nm.The mobile phase was acetonitrile(A)and 0.05%formic acid aqueous solution(B).The solvent A was held at the initial condition of 7%(v/v)for 4 min then linearly increased to 35%by 1 min and maintained for 6 min,then affter returned to 7%by 1 min,and held for 2 min balance column.The flow rate was set at 1 mL/min with column temperature 25℃ for a 14 min total time.To develop a HPLC-UV method to determine a series plasma concentration of cefepime single intravenous after SHLI or its main ingredients baicalin or chlorogenic acid for a week once a day,and the pharmacokinetics parameters were calculated..2.2 The effect of SHLI combined with cefepime on renal toxicityThe SD rats were randomly divided into 16 experiment groups namely:blankcontrol group;SHL high,medium and low dose group;cefepime high,medium and low dose group;SHL high dose combined with cefepime high,medium and low dose group;SHL medium dose combined with cefepime high,medium and low dose group;SHL low dose combined with cefepime high,medium and low dose group;and there are ten rats in each group.The MDA content and the activity of T-SOD of kidney tissue homogenate,and the serum creatinine and blood uric acid,and urine protein content and the activity of NAG enzyme,were determined after intravenous injection of combination cefepime with SHLI for a week once a day,and then rat kidney pathologic histology were detected by using H-E staining.2.3 The expression of OAT-1 were deteced by immunohistochemistryThe changes of expression of OAT-1 in rat renal tubular epithelial cells on the side of basilar membrane was observed by immunohistochemistry.ResultsFirstly,a simple,stable and reliable HPLC-UV method were developed to determine the plasma concentration of cefepime,and the method was no endogenous interference.The area under the curve(AUC)of cefepime increased about30%and 50%by combination with medium dose and high dose of SHLI,respectiveing.Treatment of combination with high dose chlorogenic acid or baicalin augmented to 2 fold the AUC of cefepime.Secondly,compared with cefepime single group,the content of MDA,serum creatinine,blood urea nitrogen,urine protein and the activity of NAG enzyme are significantly increased(p<0.01)after treatment with SHLI,indicating that treatment of combination SHLI with cefepime could caused renal injury.Thirdly,treatment of combination cefepime with SHLI could impaired renal tissue.Compareing with blank control group rats,renal interstitial hyperemia and inflammatory cells infiltration,renal tubular and glomerular cells apoptosis and malpighian protein cast were founded in other rats,which were treated by combination cefepime with SHLI.Lastly,the expression of Oatl was up-regulated in treament groups affter intravenous injection SHLI combination with cefepime,and the expression was dose dependent phase.ConclusionsTreatment of combination with SHLI and cefepime may enhance the bioavailability of cefepime,resulting in renal toxicity.The mechanism of this effect might be involved in up-regulated expression of Oatl.