Role Of Lipsomal Clodronate On Apoptosis Of Kupffer Cell In Rat With Acute Necrotizing Pancreatitis

Objective:Acute necrotizing pancreatitis (ANP) is a dangerous disease, it has rapid progression and high mortality, and it is often accompanied with systemic inflammatory response syndrome (SIRS), serious infections, septic shock and multiple organ dysfunction syndrome(MODS), finally which lead to death of patients. With the deep study on the pathogenesis of ANP mechanism, it is demonstrated that macrophages play an important role in acute necrotizing pancreatitis. This paper is to investigate the preparation of lipsomal clodronate and the role of lipsomal clodronate on apoptosis of kupffer cell in rat with acute necrotizing pancreatitis.Methods:Lipsomal clodronate was prepared by means of thin film,the model of ANP was established by injection of 5% sodium taurocholate of 4ml/kg into the pancreaatic capsule the kupffer cells were obtained from ANP rat.After exposure to different volume of lipsomal clodronate (50,100,150μL),then the proliferation and apotosis of KC was measured by MTT,flow cytometry and agarose gel electrphoresis of DNAResult:The prepared lipsomal clodronate had a suitable encapsulation efficiency of clodronate(5.8%)with an average size of 200nm, the spherical shape of liposome was confirmed by transmission electron microscope. The KC had a normal shape, the purity was more than 90 percent determined by Wright's staining, trypan blue staining found that the rate of cell survival was more than 95 percent. When exposed to different volume of lipsomal clodronate for 24h,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)% in three different volume, respectively, the difference was statistically significantly(P<0.01), the DNA of KC began degradation and gradually showed clear and characteristic ladder.Conclusion:We found that it was feasible to prepare lipsomal clodronate by means of thin film, and we prepared lipsomal clodronate successfully, it had an average size confirmed by transmission electron microscope.We found that liposomal clodronate could induce apoptosis and suppress the growth of kupffer cells in ANP rat. This study provides a new theoretical basis for the clinical treatment of ANP.