ABSTRACT
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.
ABSTRACT
Objective To study the effects of elodronate-liposome on inducing apoptosis of alve-olar macrophages from rats with acute neetotizing pancreatitis (ANP).Methods The AMs of eight rats with ANP were isolated, purified then incubated from broehoalveolar lavage by the differing rates of attachment of the various cell types in a forty-well cell culture plate.Then they were randomized in-to five groups including control group,blank liposome group( 50 μ1, 100 μ1),clodronate-liposome group (50μ1,100μ1).Values of OD were determined by MTT.AO fluorescence and haematoxylin dye were employed to determine the apoptosis of the AMs.Results There were no significant differences be-tween control group and blank liposome group(50 μ1, 100 μ1).Significant differences were found be-tween control group and clodronate-liposome group(50 μ1, 100 μ1).There were no marked differences between blank liposome group(50μ1, 100 μ1)and clodronate-liposome group(50 μ1,100μ1).AO fluo-rescence and haematoxylin dye were available to define the apoptosis of the AMs.Conclusion Clodr-onate-liposome can effectively induce the apoptosis of the AMs.
ABSTRACT
Objective To investigate the apoptosis of Kupffer cell (KC) induced by lipsomal clodronate in rat with acute necrotizing pancreatitis (ANP). Methods Lipsomal clodronate was prepared by means of thin film, the model of ANP was established by injection of 5% sodium taurocholate of 4 ml/kg into the pancreatic capsule. The Kupffer cells were obtained from ANP rat. After exposure to different doses of lipsomal clodronate (0, 50, 100, 150 μl) , then the proliferation and apoptosis of KC was measured by MTT, flow cytometry and agarose gel electrophoresis of DNA. Results The prepared lipsomal clodronate had an average size of 100～200 nm, the spherical shape of liposome was uniform and confirmed by transmission electron microscope. When exposed to different concentration of lipsomal clodronate for 24 h, the growth suppression rate was 17. 4% , 24. 2% and 31. 1% , respectively, while the apoptosis rate of the KC was (14. 12 ±0.37)% , (18.74±0.43)% and (27.51 ±0.39)%, respectively; the difference was statistically significantly (P<0. 01) , the DNA of KC began degradation and gradually showed clear and characteristic ladder. Conclusions Lipsomal clodronate could induce apoptosis and suppress the growth of Kupffer cells in ANP rats.