TITLE Studies On The Toxicity, Pharmacodynamics And Quality Control Method Of SHENKANGNING Capsules

Objective:To observe acute toxicity of SHENKANGNING Capsule in mice and long-term toxicity of SHENAKANGNING Capsule in rats. Explore SHENKANGNING Capsule improve renal function and ralated blood and urine indicators in the rats that have kidney damages induced by gentamicin(GM) Determine the content of AstragalosideⅣ in SHENKANGNING Capsule by HPLC in order to establish the method of quality control.Methods:The animal acute toxicity tests will make capsule suspension which has0.4g/ml equivalent dose, each administer the drug calculated0.4ml/10g weight to mice three times a day,at intervals of6hours.Calculate the maximum tolerated dose by three times total amount. Observe for7days after administration, record animal reactions and weight.Long-term toxicity tests in rats spent Capsule will participate in the original powder paired suspension, respectively, in low doses [1.6g/(kg·d)] and high dose [5g/(kg·d)] administered to rats3times a day, straight3months to observe the rats hematological indicators (including the clotting time RBC, WBC; platelet count, WBC and hemoglobin content), urine routine, blood biochemical parameters (AST, ALT, BUN, Cr, GLU, TCH) and the various organs (heart, liver, spleen, lung, kidney, adrenal gland, thyroid, testes or ovaries) and to carry out histopathological examination. In terms of pharmacodynamics, use the models of rats with acute renal injury by100mg/kg subcutaneous injection of gentamicin, give SHENKANGNING Capsule suspension of high (324g/L) and low-dose (108g/L)3times a day, establish the model group and the blank control group for four groups,observe serum creatinine(Cr), blood urea nitrogen(BUN), total protein (TP) and albumin (ALB) changes of each group after28days.Observe the changes of urinary protein, glucose,ketones;renal in each group. Utilize HPLC to test the content of AstragalosideⅣ in SHENKANGNING Capsule with TIANHE Kromasil C18(200mm×4.6mm,5μm) column, mobile phase:methanol-tetrahydrofuran-0.2%triethylamine solution (90:4:6)); flow rate of0.8mL·min-1, detection wavelength230nm; and the temperature of the column was30℃, injection volume is20μL Results:Acute toxicity experiment in mice showed that on the first day, the mice’s walking time and the numbers of citing the forefoot were reduced. On the second day, the spontaneous activities and food intake were both normal in the mice without any death.Caculate the maximum tolerate dose is equivalent to125times the dose of man (daily dose is12g). Compared with the control group, long-term toxicity tests in rats showed that participants spent two high dose capsules against weight gain and general behavior of rats had no effect on activity; three hematology, urine and blood chemistry were measured no significant change. The major organs in rats treated group index and the control animals no significant difference. In terms of pharmacodynamics, high and low-dose of SHENKANGNING Capsule cound significantly reduce the total protein, glucose, ketone and recovery PH in the urine of the rats that have acute kidney injury caused by gentamicin. Compared with the control group there was no significant difference (P>0.05), significantly reduce BUN and Cr content in renal injuried rat models, compared with the control group there was no significant difference (P>0.05); significantly improve the total account of the serum protein and albumin in renal injury rat models(P<0.05).Low-dose group could also improve the organ appearance symptoms such as the mast,the oozing blood,the erythema appeared among the cortex and medulla in the renal injury rats,however high-dose group has not above-mentioned phenomenon. In the range of0.1031～1.2372mg/ml, there is a good linear relationship between the concentration and peak area of AstragalosideⅣ. Linear equation Y=521559.44C-1356.36.The average recovery was99.82%, RSD was0.29%(n=9).The average content of the sample is0.3524mg/capsules.Conclusion:SHENKANGNING Capsule was administrated in the dose of1.6and5g/(kg-d)3months,the weight,urine routine,haematological indices,blood biochemical parameters,and tissue pathology of the rats had no obvious influence, equivalent to125times the clinical dose in mice given had no acute toxicity,we were able to conclude that SHENKANGNING Capsule has no toxicity. In terms of pharmacodynamics, SHENKANGNING Capsule could effectively reduce the level of CR, BUN in the acute renal injuried rats, thus inhibit the formation of hyperuricemia, and reduce uric acid salt damage to the kidneys. SHENKANGNING Capsule could also reduce glomerular filtration of protein and blood glucose, reduce glomerular hyperperfusion in the renal injuried rats, and restore damaged tissue, recovery plasma protein levels.The method of HPLC used in the article is firstly benzoylate AstragalosideⅣ and use TIANHE Kromasil C18(200mm×4.6mm,5μm) column as stationary phase; methanol-tetrahydrofuran-0.2%triethylamine solution (90:4:6))as mobile phase,detect AstragalosideⅣ in230nm wavelength.This method is simple and accurate for determination of amygdalin in SHENKANGNING Capsules.