聚乳酸乙醇酸／RNAⅢ抑制肽缓释微球的血液相容性

目的：研究表明RNA Ⅲ抑制肽（RNA Ⅲ inhibiting peptide，RIP）是葡萄球菌一种有效的全面抑制剂，作用机制与传统抗菌素明显不同，有着良好的应用前景。通过实验评价聚乳酸乙醇酸（polyaitlcglycolic acid，PLGA）／RIP缓释微球的血液相容性。方法：实验于2005-10／2007-10在解放军总医院临床药理研究所及医学动物实验中心完成。选择成年健康新西兰大白兔30只，按随机数字表法分组，每组6只。药品及试剂：PLGA，二甲基亚砜、MTT，DMEM，二硝基氟苯。实验方法：①）PLGA／RIF微球制备：采用Fmoc法由C端至N端先合成粗品肽：采用反相液相色谱法对RIP粗品进行纯化分析，按紫外吸收峰收集组分，冷冻干燥，得到RIP纯品。再采用液相复乳法制备直径50-70mm的PLGA／RIP微球。②洗提液制备：PLGA／RIP微球粉末按1g／L在37℃无菌条件下用生理盐水洗提72h，制得洗提液原液；加入同体积的无菌生理盐水制得0．5g／L的稀释液。③溶血实验：以蒸馏水和生理盐水分别为阳性、阴性对照，观察PLGA／RIP洗提液原液和0．5g／L洗提液的溶血率。④凝血实验及PLGA／RIP对凝血酶原时间和活化部分凝血酶时间的影响实验：以生理盐水为阴性对照，观察PLGA／RIP洗提液原液和0．5g／L洗提液对兔凝血时间的影响和凝血酶原时间和活化部分凝血酶时间的影响。⑤PLGA／RIP对兔白细胞、红细胞和血小板及血小板聚集的影响实验：观察PLGA／RIP洗提液原液和0．5g／L洗提液对兔白细胞、红细胞和血小板及血小板聚集的影响。结果：纳入动物30只。均进入结果分析。①溶血实验结果显示PLGA／RIP洗提液原液和0．5g／L洗提液的溶血率分别为3．24％和2．67％，两者的溶血率均〈5％，符合医用生物材料的溶血实验要求。②凝血实验结果表明PLGA／RIP洗提液原液和0．5g／L洗提液对兔凝血时间无明显影响?

Blood compatibility of polyaiticglycolic acid/RNA Ⅲ inhibiting peptide microspheres

AIM： Studies reveal that RNAⅢ inhibiting peptide （RIP） is an effective suppressant for staphylococci and the mechanism obviously differs from traditional antibiotic drug, indicating its potential application. This study evaluated blood compatibility of polyaiticglycolic acid （PLGA）/RIP microspheres. METHODS： The experiment was performed in the Pharmacological Research Institute and Animal Experimental Center at the General Hospital of Chinese PLA from October 2005 to October 2007. Thirty healthy adult New Zealand white rabbits were grouped randomly, with 6 animals in each group. Drugs and reagents： PLGA, dimethyl sulphoxide, MTT, DMEM, dinitoflruorobenzene.1. Preparation of PLGA/RIP microspheres： Fmoc method was used to synthesize RIP from C end to N end, then the synthesized crude peptide sample was purified by the reverse phase high performance liquid chromatography, and composition was collected by means of ultraviolet absorption peak. The purified PIR was obtained from freezing and drying. Afterwards liquid-phase multiple emulsion method was used to synthesize PLGA/RIP microspheres of 50-70 mm diameter.2.Preparation of eluent： 1 g/L PLGA/RIP microsphere was eluted sterily with saline at 37℃ for 72 hours to harvest stock solution, then equal volume of stroke-physiological saline solution was added to obtain 0.5 g/L dilution.3.Haemolysis test： With distilled water and stroke-physiological saline solution as positive and negative controls respectively, the rate of haemolysis of 100% and 50% eluent of PLGA/RIP were studied.4. Hemagglutinatin test and PLGA/RIP effect on prothrombin time （PT） and activated partial thromboplastin time （APTT）： With stroke-physiological saline solution as negative control, the effects of 100% and 50% eluent of PLGA/RIP on PT and APTT were observed.5.The effects of 100% and 50% eluent of PLGA/RIP on the leucocyte, erythrocyte, thrombocyte and platelet aggregation were detected. RESULTS： Totally 30 rabbits were involved in the result analysis.1.Haemol