The Protective Function Of Baoshentang For Acute Renal Failure

Renal ischemia is one of the most common causes of acute renal failure (ARF), initiating a complex and interrelated sequence of events, resulting in injury to and the eventual death of renal tubular cells. But the mortality associated with ARF has not decreased substantially over the last 50 years. Objective: We designed a series of experiments to find out new prescriptions to reduce the renal dysfunction and injury of the rat kidney caused by ischemia/reperfusion (I/R) and to be used in clinical prevention and therapy later. Methods: 1.HK-2 cells were seeded for 1-12 days in 96-well plate at the concentration of 1*105 cells/ml, then OD(optical density) was measured by the method of MTT. 2. HK-2 cells were treated for Oh 2h 4h 6h 8h lOh 12h with glucose deprivation, a mitochondrial inhibitor or a combination, then OD was measured. 3.More than 30 extractions of Chinese herbal drugs were examined in this study and each was administered into cell culture for Ih, then in chemical hypoxic medium for 4h and subsequently OD was measured. The immunocytochemistry of anti-HSP70 in vitro was processed in the cells pretreated with Quanshen, Longdancao, Duhuo, IBE5 and Diyu. 4. Every two single drugs were administered together into cell culture for Ih, then in chemical hypoxic medium for 4h and OD was measured. 5.Male Wistar rats were randomly divided into eight groups(six rats per group), five groups administered Longdancao, IBE5, Baihuasheshecao, Duhuo (50mg each) and L-argiriine (40mg) 1h before undergoing bilateral renal ischemia(45 min) followed by reperfusion(6h and 24h). One group was administered Shenweitang (3ml twice a day for three days) before ischemia. Plasma concentrations of urea and creatinine weremeasured for the assessment of renal dysfunction and I/R injury at the 24-h time point. Kidney myeloperoxidase (MPO) activity and malondialdehyde(MDA) levels were measured for assessment of neutrophil infiltration and lipid peroxidation at 6-h time point, respectively. Renal sections were used for histologic grading of renal injury and for immunohistochemical localization of inducible nitric oxide synthase (iNOS) at 24-h time point. Results: I.We obtained the growth curve of HK-2 cells, from which we conclude that the fifth day was appropriate for the test by MTT method. 2. A chemical hypoxic model was successfully set up to screen the drugs in vitro. 3. 19 effective drugs and 61 effective combinations of two single drugs against hypoxia were screened out based on the hypoxic model. The immunocytochemistry of HSP70 in vitro showed that pretreatment with Quanshen, Longdancao, Duhuo, IBE5 could enhance the expression of HSP70, except for Diyu. It indicated that the protective mechanism of the four drugs might be relative to it. But the mechanism of Diyu should be proved later.4. Shenweitang showed obvious protection to renal tubular cells in vivo, which might be relative to many mechanisms, such as the inhibition of neutrophil Infiltration, the reduction of ROS (Reactive Oxygen Species) production, and the selective blockade of iNOS. Conclusions: Shenweitang can protect the kidney from being injured caused by I/R, thus it is hopeful to be used to prevent and treat the acute renal failure caused by I/R.