Effects Of RhaFGF On Promoting Rehabilitation Of Skin Ulcerations And The Distribution And Excretion Of RhaFGF In Rabbits

Part one Effects of rhaFGF on promoting rehabilitation of skin ulcerations in rabbitsObjective To study the effects of recombinant human acidic fibroblast growth factor (rhaFGF) on promoting rehabilitation of skin ulcerations in rabbits.Methods The skin ulceration models of rabbits were made and divided into groups, among which two groups received 90, 60 AU·(cm2)-1 rhaFGF, the positive control group received 60 AU·(cm2)-1 rb-bFGF, and the negative control group received only the base of the formulation of the equal quantity as the second group. The ulceration area was recorded by photos and cellophane film at 1,4,8d after ulcerations, and the volume of injured cavity was measured with water injection. The reduced degree of the area and the volume were compared. At 8,14d after ulcerations, the growth of wound granulation and re-epithelization were tested to evaluate the effects of rhaFGF.Results The skin ulcerations of the rabbits became smaller as time went on in all groups, but 60 AU·(cm2)-1 rhaFGF and 60 AU·(cm2)-1 rb-bFGF groups are more significantly than that in the negative control group. The volume of injured cavity also grew smaller, and 60 AU·(cm2)-1 rhaFGF and 60 AU·(cm2)-1 rb-bFGF groups are more significantly. At 8d after ulcerations, fibroblasts and capillary granulation tissue grew more and actively, in 60 AU·(cm2)-1rhaFGF and rb-bFGF groups more significantly than that in the negative control group. At 14d after ulcerations, the ulceration contracted and re-epithelized in 60 AU·(cm2)-1rhaFGF and rb-bFGF groups more significantly than that in the negative group.Conclusion rhaFGF can promote rehabilitation of the skin ulcerations in rabbits by promoting the growth of granulation tissue, capillary angiogenesis and epithelium.Part two Study on the distribution and excretion of 125I-rhaFGF after it was given to the rabbits intravenously.Objective To observe the distribution and excretion of 125I-rhaFGF after it was given to the rabbits intravenously offering information about parent-drug's distribution in the serum after local application.Methods 1) rhaFGF was radiolabelled with 125I and its radiochemical purity (RCP) measured by high performance liquid chromatography (HPLC). 2) 125I-rhaFGF(13μg·kg-1) was given to 15 rabbits intravenously. At 5min,15min and 2h after injection, 5 rabbits were killed separately and the liver, kidney, lung, myocardium, thyroid gland and other organs were obtained and homogenized. Meanwhile, the serum, bone marrow and urine were got, too. Then, TCA solution was added to them separately to deposit them for measuring the total and TCA sedimentum's radioactivity. 3) 125I-rhaFGF(5μg·kg-1) was given to 5 rabbits intravenously. At 2h,8h,12h,24h,36h and 48h after injection, the urine and dung were collected for measuring the radioactivity to calculate the percentage of the excreting radioactivity occupying the injecting radioactivity. 4) After centrifuging the collected urine, the supernatant was obtained to separate by HPLC, and the chromatogram was compared with the 125I-rhaFGF standard chromatogram.Results 1) The RCP of 125I-rhaFGF was 98.13±0.57%. 2) The distribution of the total and TCA sedimentum's radioactivity at different time point after the rabbits were injected 125I-rhaFGF intravenously was documented. According to the area under the time-concentration curve (AUC), bone marrow, femur, liver, kidney, spleen and suprarenal gland's radioactivity was highest, and that of serum was relatively lower, however, genital system, muscles and brain were lowest. 3) The urine's total radioactivity was higher relatively, but its TCA sedimentum's radioactivity was lowest. 4) The serum's ratio of the TCA sedimentum's radioactivity and the total radioactivity was decreasing by the time. 5) Before 12h, the radioactivity excreted faster by urine, 68.07±8.99% of the injecting radioactivity, but more slowly after 12h. At 48h, the urine's excreting quantity was 80.82±2.82% of the injecting radioactivity, yet the dung's was 0.61±0.61%. 6) According to the HPLC analysis, there were 125I hydrophilic products of metabolism in the urine mainly.Conclusion 1) The distribution of 125I-rhaFGF has target after whole body absorbing. 2) The urine's radioactivity is mainly 125I products of metabolism. 3) The parent-drug's ratio decreased by the time. 4) 125I-rhaFGF excreted mainly by urine, little by dung, and before 12h excreted more quickly. 5) There are 125I hydrophilic products of metabolism in the urine mainly.