| Dishen granular,composed of Schisandra chinensis Fructus,Rhubrab,Rhubrab Carbon,Astragalis Radix,Radix Salviae Miltiorrhizae,Cattail Pollen carbon etc,is a precompounded prescription in medical institute.The granular has the effect on tonifying kidney,activating blood,smoothing Qi and expelling toxin,so that,it regulates a series of symptoms and metabolic disorder caused by chronic renal failure in clinic.In this experiment,the pharmacodynamics of Dishen granular was studied by pharmacodynamics test.To provide experimental basis and theoretical basis for the development of new drug,quality standard and stability Dishen granular are studied on the basis of the active ingredients and the national pharmacopoeia standard of the Chinese herbal medicines.The constituents of the prescription were identified by TLC,and the content of each constitute was determined by HPLC.Aim:The purposes of this research were to study the influence of Dishen granular on the sensitive proteins KIM-1 and NGAL in kidney ischemia-reperfusion rats.What is more,this study was done to establish a method for the identification of effective constituents and the measurement of each content in the Dishen granule.Methods:The experiment was proceeded by dividing SD rats into 5 groups,including sham operation group,model group,Niaoduqing Granule positive group,high dose group and low dose group of Dishen granular.In addition to the sham operation group,all the other groups underwent bilateral renal ligation and reperfusion,which resulted in acute ischemia-reperfusion injury.Continuous medication for 1 day,3 days,7 days after the operation.Serum urea nitrogen(BUN)and creatinine(SCr)were detected,the renal pathological morphology was observed,KIM-1 and NGAL of kidney were detected by Western Blot.Furthermore,the constituents of the prescription were identified by TLC,and the content of each constitute was determined by HPLC.Results and conclusion:After treated with Dishen granular,Serum urea nitrogen(BUN)and creatinine(SCr)levels and the level of NGAL and KIM-1 in renal tissue of model group rats increased significantly(P<0.001).During day 1,3 and 7,compared with the model group,the level of serum BUN and SCr of Dishen granular high dose group and low dose group were significantly reduced(P< 0.001,P<0.05),the protein expression of NGAL decreased significantly(P<0.001),however,KIM-1 protein expression decreased but with no statistical significance.HE staining showed that Dishen granular can reduce the occurrence and development of renal tubular epithelial cells,tubular cell necrosis,bleeding and inflammatory reaction,the effect of Dishen granular high dose was more dramatic.The conclusion that Dishen granular protect the kidney by reducing the expression of KIM-1 and NGAL was drew.The establishment of Dishen granular quality standard illustrated the method for the determination of the content,Schisandra,in Dishen granular.The results indicated that the calibration curve showed a good linear correlation in the range of 0.552~5.52?g(r=0.9993),and the average recovery of Schisandra was 98.79%,RSD=1.58%.The average contents of Schisandra in the three batches of granules were 2.85 mg,2.86,Mg and 2.85 mg,respectively.The TLC identification methods of Radix Salviae Miltiorrhizae?stragalis Radix?Rhubrab?Cattail Pollen carbon were proved simple,accurate,sensitive and specific,which provided an experimental basis for controlling the quality of Dishen granules and for reporting new drugs.The results of preliminary experimental study on stability of Dishen granular showed that,with high temperature and high humidity,illumination,sample observation and accelerated test,three batches were no moisture absorption and deliquescence and agglomeration phenomenon,TLC identification and content determination are in conformity with the provisions,no obvious change,good preparation stability,quality control. |
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