[Study of immunolipo-sodium morrhuate applied on human hemangioma endothelial cells in vitro].

PURPOSE: To explore the method of preparing immunolipo-sodium morrhuate and evaluate its effect on human hemangioma endothelial cells in vitro. METHODS: Using SPDP((N-Succinimidyl-3-(2-pyridyldithio)) propionate) as cross-linker, anti-VEGFR2/KDR monoclone antibody was combined to the liposome surface to prepare immunolipo-sodium morrhuate by extruding method, and then its effect on human hemangioma endothelial cells in vitro was observed by laser scanning confocal microscope, inverted microscope, Gimsa staining, transmission electron microscope, MTT and flow cytometry. RESULTS: The average diameter of the immunoliposome was 122.9 nm, which had a very good stability when compared with normal liposome, it had stronger and faster combining ability, its potential to induce apoptosis was much more prominent, and its toxic effect on human hemangioma endothelial cells was gentle, which was similar to normal liposome. CONCLUSION: We have prepared immunolipo-sodium morrhuate successfully, which has very good specific initiative targetting ability in vitro and can induce pervasive apoptosis of human hemangioma endothelial cells.

[The experimental study of lipo-sodium morrhuate promoting apoptosis of ECV-304 cells].

OBJECTIVE: To evaluate the effect of lipo-sodium morrhuate on ECV-304 cell line. METHODS: The effect lipo-sodium morrhuate was evaluated by toxicology trial (MTT), electron microscope, DNA electrophoresis and flow cytometer. RESULTS: The toxicology results showed, that the number of vital cells in lipo-sodium morrhuate group decreased slowly. The electron microscope exhibited apoptosis in the lipo-sodium morrhuate group. And there were typical DNA ladder in DNA electrophoresis and typical apoptosis peak in flow cytometer. The apoptosis rate was 22.23%. CONCLUSIONS: Unlike the normal preparation of sodium morrhuate, lipo-sodium morrhuate could induce apoptosis of ECV-304 cell line.

[Compare the effect of lipo- and normal prepared Sodium Morrhuate on ECV-304 cell line].

OBJECTIVE: To compare the effect of Sodium Morrhuate on ECV-304 between its lipo- and normal preparation. METHODS: The ECV-304 cell line was supplemented with Sodium Morrhuate and lipo-Sodium Morrhuate in order, and the result on morphology (microscope, Giemsa Staining and electron microscope), cell activity (MTT), and flow cytometer between the two preparation were compared. RESULTS: In normal preparation group, cell's edema occurred. Chromatin was like catkins. Tumefaction and degeneration of mitochondrion and endoplasmic reticulum appeared. In lipo-Sodium Morrhuate group, the membrane was creased and processus appeared. Chromatin aggregates to the membrane of nucleus was like crescent, and then broken. The apoptotic body was formed. MTT changes showed that the curve of the normal preparation group was steep and the change time was short relatively, which cues the vital cells decreased sharply. The curve of lipo-Sodium Morrhuate group was gentle and the change time was long relatively, which cues the vital cells decreased slowly. The flow cytometer showed that typical apoptosis peak appeared. CONCLUSION: The normal preparation group shows an acute toxic effect on ECV-304 cell line, which result in a necrosis course, while lipo-Sodium Morrhuate shows a gradual releasing process, which may indicate a apoptosis course.