Clodronate脂质体的制备及诱导重症急性胰腺炎大鼠腹腔巨噬细胞凋亡的实验研究  

Preparation of lipsomal Clodronate and its inducing effect on apoptosis of peritoneal macrophages in rats after severe acute pancreatitis

作　　者：蔡华忠[1] 张建新[1] 尹江涛[1] 崔磊[1] 党胜春[1] 

机构地区：[1]江苏大学附属医院,镇江212001

出　　处：《中华肝胆外科杂志》2010年第12期943-946,共4页Chinese Journal of Hepatobiliary Surgery

基　　金：国家自然科学基金资助项目（30772117）;江苏省自然科学基金资助项目（BK2007096）;镇江市社会发展基金项目（SH2008036）

摘　　要：目的 探讨clodronate脂质体的制备及其对重症急性胰腺炎（severe acute pancreatitis,SAP）大鼠腹腔巨噬细胞（peritoneal Macrophages,PM）凋亡的诱导作用.方法 利用薄膜法制备clodronate脂质体;用5%牛磺胆酸钠逆行胰胆管注射制备SAP模型,分离培养SAP大鼠PM.PM经不同剂量的clodronate脂质体（50、100、150μl）作用后,用MTT法检测其对PM增殖的影响.用Annexin V/PI双标记法和DNA琼脂糖凝胶电泳技术检测PM凋亡情况.结果 制备的脂质体直径200 nm,clodronate包裹率为5.8%.透射电镜检查见其大小均匀.在3种不同剂量clodronate脂质体干预下,MTT结果显示巨噬细胞的生长受到明显抑制（P＜0.01;流式细胞术检测显示,在3种不同剂量clodronate脂质体干预组,PM凋亡百分率分别为（10.32±0.34）%、（18.16±0.49）%和（29.87±0.35）%,差异均有显著性（P＜0.01）.1.2%琼脂糖凝胶电泳显示,随着clodronate脂质体剂量的增高出现渐清晰的特征性梯状条带.结论 clodronate脂质体可诱导SAP大鼠PM的凋亡.Objective To explore the preparation of liposomal clodronate and investigate its inducing effects on the apoptosis of peritoneal macrophages in rats after severe acute pancreatitis （SAP）.Methods Liposomal clodronate was prepared by means of thin film. SAP rat model was established by retrograde injection of 5% sodium taurocholate into the pancreatic duct. The peritoneal macrophages were obtained from SAP rats. After exposure to different doses of liposomal clodronate （50, 100,150 μl）, the PM proliferation was determined by MTT colourimetry. The apoptosis of PM was measured by flow cytometry and agarose gel electrophoresis, respectively. Results The prepared liposomal clodronate had a suitable encapsulation efficiency of clodronate （5.8%） with an average size of 200 nm. The spherical shape of liposome was confirmed by transmission electron microscope. Exposed to liposomal clodronate of different doses resulted in a obvious growth depression （P〈0.01）. The apoptotic rate of the PM was （10.32±0.34） %, （18.16±0.49）% and （29.87±0.35）% in three different dose groups and the difference was marked （P〈0.01）. 1.2% of agarose gel electrophoresis of DNA extracted from apoptotic macrophages induced by liposomal clodronate showed clearer and characteristic ladder following the liposomal clodronate concentration. Conclusion Liposomal clodronate has a definite effect on peritoneal macrophages in SAP rats.

关 键 词：急性胰腺炎 巨噬细胞 凋亡 Clodronate脂质体 

分 类 号：R686[医药卫生—骨科学]