Promote Hepatocyte Growth Factor Precursor Liposomes

Average molecular weight of promoting hepatocyte growth factor (PHGF) was 12.1 kD determined by SDS- polyacrylamide gel electrophoresis method (SDS-PAGE). Isoelectric point of PHGF was at a range from 6.3 to 6.6 determined by isoelectric focusing electrophoresis method (IEF). Chemical stability of PHGF water solution was measured by HPLC method, which indicated that the solution should be stored at low temperature and against light. Results above provided basis for designing formulation, establishing separation and analysis method, and studying stability of PHGF proliposomes. UV method was firstly set to determine total protein contents of PHGF proliposomes. Compared with Lowry method, UV method overcame many disadvantages such as complicated operation and many interfering factors, and eliminated the effects of ingredients. This method combining with gel filtration was used to determine encapsulation efficiency of PHGF liposomes. PHGF proliposomes were prepared by dried-reconstituted vesicles method. Membrane composition, media pH value and preparation process were studied. On the base of single factor test, uniform design was used to obtain the optimum component of proliposomes and preparation process; the best formulation had been identified, which raised encapsulation efficiency from 5% to 25%. This method could resolve the problems of chemical stability of biochemical drug and physical stability of liposomes. Studies on stability indicated that PHGF proliposomes should be stored at low temperature and against light, which ensured unchange of their redispersity, content of drug, encapsulation efficiency, acid value and peroxide value. Liposomes were multilamellar vesicle structure under transmission electron microscopy. Average particle size was 0.65 micrometer, ranging from 0.09 micrometer to 1.21 micrometer. Cryoscopic value was 0.64 C determined by cryoscopic method, which was a little higher than that of isotonic solution. Zeta potential was -44.23 millivolt. Viscosity was 1.83 mpa. All of which were coincided with physical desire. In addition, the tests of PHGF liposomes including irritation, hemolysis, pyrogen, anomaly toxicity and hypersensibility, proved that the preparation reached the standard of intravenous injection entirely. Taking glutamic pyruvic transaminase (GPT) and malondialdehyde (MDA) in mice as index, studies on pharmacodynamics after hepatic injure caused by tetrachloromethane in mice indicated that GPT level (P<0.001) and MDA yieldment (P