Cl₂MBP-Liposome Is Prepared And Study Apoptosis Of Macrophages Induced By Cl₂MBP-Liposome

Objectives One of the aims of the study is to prepare the vehicle of Cl₂MBP. Another is to explore apoptosis of macrophages induced by Cl₂MBP-liposome and offer the new method and proof which will help study the function of macrophages in vivo and cure some diseases in clinic.Methods In our laboratory, a conventional method is used for the production of lipsome which encapsulate Cl₂MBP, under low vacuum and rotation evaporation. In the end, a thin milky white phospholipid film is formed against the inner wall of the flask. The phospholipid film is dispersed with different aqueous solution (PBS, Cl₂MBP) by gentle rotation to prepare empty liposome and Cl₂MBP-liposome. Monoclonal antibody is used to detect the apoptosis of macrophages induced by Cl₂MBP-liposome in vivo and vitro. Agarose gel electrophoresis of DNA was examined apoptosis of macrophages.Results The liposome prepared by conventional method can encapsulate Cl₂MBP and deliver Cl₂MBP to macrophages directionally. Cl₂MBP-liposome induces the apoptosis of macrophages.Cl₂MBP-liposome incubates peritoneal macrophages for 0, 3, 12, or 24 hours in vitro. After being incubated for 0 hour, no apoptotic cell is found ,after 3 hours, the characteristic apoptotic cells show macrophages with brown nuclei, while non-apoptotic macrophages with blue nuclei detected with monoclonal antibody at this time. The percentage of apoptosis is 26% after 3 hours, 32% after 12 hours and 43% after 24 hours.Peritoneal macrophages are collected and coated on the glass slice...