| Objective:1. To observe the properties of Saikosaponin D(SSd) pharmacokinetics in mice.2. To make it clear whether SSd has significant interaction with anticancer drugs when they are administrated.3. To explore the inhibition effect of SSd in xenografts derived from MCF-7/ADM cells in nude mice.Method:1. Study of the pharmacokinetics of SSd in AnimalKunming mice were selected as the study model for pharmacokinetic study. SSd(5 mg/kg) were given by intraperitoneal injection to mice,the blood plasma was collected at different time. The content of SSd in the blood plasma was detected by HPLC at different time. The extracts of samples were chromatographed on a ZORBAX SB-C18 column. HPLC condition: the mobile phase consisting of water: acetonitrile(62:38), 25℃column temperature, the mobile phase pumped at a flow rate of 1m L/min, 8nm bandwidth, 205 nm of ultraviolet detection.2. Influence of SSd on Adriamycin PharmacokineticsKunming mice were divided into 3 groups by weight of mice at random,four mice as a group, male and female half 2 only. Each group of mice wasadministrated at different time point respectively. SSd group was given SSd through intraperitoneal injection at the dosage of 5mg/kg. ADM group was given adriamycin by tail intravenous injection at the dosage of 5mg/kg. SSd+ADM group was given SSd through intraperitoneal injection at the dosage of5mg/kg and adriamycin by tail intravenous injection at the dosage of 5mg/kg respectively. The animal blood samples were collected at different time point,and then plasma was separated. HPLC method was adopted to detect adriamycin contents in plasma. We estabolished the chromatographic detection method for blood drug concentration of adriamycin. The linear concentration range,sensitivity, precision and recovery rate and other indexes meet the testing needs.3. Inhibition effect of SSd in xenografts derived from MCF-7/ADM cells in nude miceThe inhibition rate of MCF-7/ADR and MCF-7 cell proliferation was measured with a MTT assay. The resistance index was calculated. Established adriamycin resistanee human breast cancer models that xenograft transplanted subeutaneous in nude mice. Randomized the mice into 4 groups: control group,SSd group, ADM group and SSd+ADM group. After the models were established, different treatments were given respectively: control group was treated with saline water 0.2 ml intraperitoneally, SSd group was treated with SSd 5mg/kg intraperitoneally, ADM group was treated with adriamycin 5mg/kg intraperitoneally, SSd+ADM group was treated with SSd 5mg/kg andadriamycin 5mg/kg intraperitoneally every two day. The general status of nude mice was observed and tumor growth was monitored from the first day of treatment. The long diameter and short diameter of the xenograft were measured before treatment every two day. The tumor volumes were calculated, growth curve was drawn and the weight of mice before and after treatment was recorded.The mice were sacrificed on the 21 th day. Every xenograft was cutdown and weighed.Result:1. After 5mg/kg SSd administered intraperitoneally to mice at the different time point, SSd content in the blood plasma was detected. The results showed that the SSd accumulation in plasma reached the peak value at 4 h(954.7 ± 36.2ng/ml), and then the concentration gradually reduced. The blood plasma concentration-time course fitted well to two compartment model according to the pharmacokinetic statistical software analysis.2. Parameter assessment was calculated by pharmacokinetics statistical software. The results showed that ADM concentration-time course in plasma also fitted well to two compartment models. The difference was not significant in statistics between ADM group and SSd+ADM group.3. The xenograft volume of both ADM group and ADM+SSd group was smaller than that of control group after the 3 weeks’ treatment, with statistical significance(P<0.05). The xenograft volume of ADM+SSd group was smallerthan that of ADM group and SSd group, both with statistical significance(P<0.05).Conclusion:1. 5mg/kg SSd administered intraperitoneally to mice, the blood plasma concentration-time course fitted well to two compartment models, and SSd concentration in vivo can achieve effective concentrations of reversing multi-drug resistance in vitro.2. The pharmacokinetic characteristics of ADM were not difference in the presence and absence of SSd in Kuming mice.3. Establishment of ADM resistance human breast cancer models in nude mice successfully, and MCF-7/ADM xenograft can be reversed by SSd in vivo. |
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