Pharmacodynamic & Toxicological Study On Therapy Of Occlusive Soft Tissue Injury With Injury Pain-killing Ointment

PurposeStudy antiinflammation and abirritation of the traditional Chinese medicine, Injury Pain-killing Ointment (IPO)for external application, as well as safety of application on skin; lay solid foundation for further exploring function mechanism of IPO and theoretical foundation for clinic applicationMethod1. Pharmacodynamic Study 1.1 Abirritation Experiment1.1.1. Method of making rats body quiver: Take 40 healthy white rats of Kunming Species, and divide them into four groups at random, i.e. Negative Control Group, Positive Control Group (apply Zhenjiang Gao, ZJG), Small Dose Group (apply IPO), Large Dose Group (apply IPO). Make sure each group has five males and five females, hairs on 1 .1 cm2 belly of which are shaved off. Apply different medicine for trials on them. Inject 0.6% acetic acid solution approximately 0.2ml into abdominal cavity of each rat 30 minutes later and observe frequency that rats quiver their bodies within 15 minutes and calculate obtunding rate.1.1.2. Making rats take lukewarm bath: Put a 50ml small beaker filled with lukewarm water into hot-plate pain measurer at controlled temperature of 50℃, and then make tail of rats immersed in 50℃ water, taking the time seeing tail retracting as pain threshold. Take 40 healthy white rats of Kunming Species and divide them into four groups based on their pain thresholds at random. Make sure each group has five males and five females, hairs on 1 . l cm2 belly of which are shaved off. Apply different liquors on them. Measure their pain threshold 30 minutes later and observe abirritation of medicine through change of pain threshold, which are observed respectively at the 30th, 60th, 90th and 120th Minute of experiment upon administration.1.2. Antiinflammation Experiment1.2.1. Making ear of little rats tumefy: Take 25~30g weight male rats totaling 40, and divide them into four groups. Make sure each group has ten. Apply 0.02-ml dimethylbenzene solution on each rat, and reapply medicine for trial half an hour later. Make rats die of spondylopathy dislocation four hours later, shear off both ears from auricle baseline, and make a didcus respectively on the same location of ears with Φ 9mm pin-hole plotter and weigh them with torsion balance to obtain tumescence degree, which equals to weight of left ear minus that of right ear.1.2.2. Making foot of big white rat tumefy: Select big white rats (Wistar) weighing 130-150-g and perfuse 5-10ml water to them prior to experiment. Stretch their posterior limb, use 26 # syringe meedle injector to inject some10% egg white into such limb frommid subcutaneous of metatarsus upward, and then finish injection downward, totaling 0.1 mi, meanwhile, apply different medicine for trial on such metatarsus and measure tumescence with mercury volumetric method at the 5th, 30th, 60th, 120th, 180th and 240th Minute from then on.1.3. Impact on permeability of capillary on rat skin.Divide 40 health rats into four groups, and make sure each group has five males and five females. Apply different liquors for trial on belly skin 60 minutes before experiment. Inject 0.5% Evans Blue physiological saline of 0.1ml/20g via vena one hour later, and then apply 100% dimethylbenzene solution of 0.03ml on belly skin. Kill such rats 20 minutes later, shear off a block of skin totaling 1.5x1.5 cm2, and break it into pieces, immerse them in acetone-water (7:3), perform filtering 24 hours later, then compare colors of filtrate with 751-Type Spectrophotometer (A=590nm) and measure absorbency, which indicates content of Evans Blue in skin and indicates permeability of skin capillary indirectly. 2. Safety and Toxicity Experiment2.1. Skin Irritation Experiment on rabbit: divide nine health rabbits (six males and three females) into three groups; make sure each group has two males and one female. Depilate both sides of their backbones without injuring skin and make sure depilating area is 5x10 cm2on both sides (approximately 10 % of rabbit body), use one depilated area for normal skin irritation experiment and divide it into administration p...