Experimental Study On Wound And Hypertrophic Scar Of Rabbit Ears And Animal Skin Toxic Effect Experimental Study Of "Cuyubanhenxiao"

Objective To develop new compound Chinese traditional medicine-Cuyubanhenxiao(CYBHX) by adding medicine to traditional Heibuyaogao in term of trace element; To measure the content of trace element of CYBHX and compare with drugs of promoting wound surface healing and curing scar in market sold now. To investigate the effect of the new ultra-fine attrition technique on trace elemental solubility of Chinese traditional medicine. To provide a theoretical basis for clinical application of trace element in Chinese traditional medicine to promote wound surface healing and prevent hypertrophic scar .Methods Making the total ultra-fine attrition ointment (CYBHX) with the new total ultra-fine attrition technique. Measure the content of trace element of CYBHX,DQ and recombinant human basic fibroblast growth factor. Iron, zinc, cuprum were measured by the flame atomic absorption spectrometry. Manganese was measured by the graphite furnace atomic absorption spectrometry and selenium was measured by hydrogenation atomic absorption spectrometry.Results The content of selenium can' t be detected in every drugs. The content of zinc, cuprum iron, manganese is higher than DQ and bFGF. cuprum iron, manganese can be detected in DQ and only manganese can be measured in bFGF. Conclusion The new ultra-fine attrition technique can improve trace elemental solubility of CYBHX remarkably. The content of trace element is higher than DQ and bFGF. It can provide a theoretical basis for clinical application to promote wound surface healing and cure scar in term of trace element. Objective To explore the function of "CuYuBanHenXiao" on wound and hypertrophic scar of rabbit ears. And study the acute toxicity, irritation and sensitivity effect of Cuyubanhenxiao to New Zealand big white rabbit' s nomal skin and impaired skin. To provide a theoretical basis for clinical application of "CuYuBanHenXiao" .Methods (1) Creat animal model of rabbit ear of wound surface over the ventral surface of ears in 9 adult New Zealand white rabbits. 9 rabbits were randomly divided into group A₁(CuYuBanHenXiao),group B₁(bFGF) and group C₁(saline). After the formation of hypertrophic scar, drugs were began to adopt on group A₁,B₁,and C₁ once every day for 1 month. Recording the time of wound surface and comparing the difference between the three groups. After the formation of of hypertrophic scar , measure the thickness of scar at 0d,5d, 9d, 21d, 31d and compare the long-term effect between A₁ and B₁. At the same time, 3 scars were cut in A, and B₁ to measure the content of trace element of scar and compare the difference between A₁ and B₁. (2) Creat animal model of rabbit ear of hypertrophic scar over the ventral surface of ears in 18 adult New Zealand white rabbits.18 rabbits were randomly divided into group A₂(CuYuBanHenXiao),group B₂(DQ) and group C₂(saline), each group having 6 rabbits. After the formation of hypertrophic scar, drugs were began to adopt on every group once every day for 1 month. Measure the thickness of scars at the formation of scar , 5d, 9d, 21d and 30d. At 1 week, 2 week and 4 week, operations were done simultaneously in three group, one ear scar of each rabbit was cut down every time for immunohistochemical (P53, PCNA), histomorphology and the determination of MDA,SOD and CAT . (3) 40 New Zealand big white rabbits were dividesed into 3 groups. â‘ 12 rabbits were randomized into 3 groups, each group is 4 rabbits. The medicine is Vaseline, Cuyubanhenxiao (44 %), Cuyubanhenxiao(70%). They were separately applied to A₃, B₃, C₃ 3 groups. Each rabbit with 50 cm² was molted on both sides of backbone to take the acute skin toxicity test. The right sides every group were used medicine (A₃, B₃,C₃)after 7 days, the nomal skin acute toxic effect was observed. The left sides were scratched to even hemorrhage and spreaded medicine on every group rabbit. 7 days later, the damaged skin toxic effect was observed. (2)4 rabbits were molted on both sides of backbone of every rabbit and there were 2 molted area on each side of every rabbit. The medicine group was completely same with the first group(B₄,C₄). The left side of every molted areas were spreaded by Vaseline (A₄). The right molted areas of near to the head were spreaded by B₄ and the other areas were spreaded by C₄. 7 days later, the nomal and damaged skin irritation effect was observed. (3)24 rabbits were randomized into A₅,B₅ and C₅ 3 groups, the number of each group was 8. The medicine was Vaseline, Cuyubanhenxiao(44% ), 1%2, 4-Dinitrochlorobenzol. Each rabbit with 50 cm² was molted on the left sides of backbone to take the the initial contact and the stimulation test. The sensitivity effect of medicine was observed. Results (1)There is no significant difference between A₁ and B₁ on wound healing time(P>0. 05). There is significant difference between A₁,B₁ and C₁. After the formation of scar for 1 month, the scar of A₁ group extincted faster than B₁ group. After the wound healing, the content of trace element of A₁ group was higher than B₁ group. (2) The thickness of scar in A₂ and B₂ group is obviously thinner, the color is blander , the texture is sofer, the alinement of collogen is raritaser and the color is blander lapse of time of administer than C₂ group. With the time going, the difference is more manifest ;The thickness of scar has significant difference between A-2,B₂ and C₂ group(P<0. 05). The expression of p53,PCNA has significant difference between A₂,B₂ and C₂ group between 1 week and 4 week(P<0. 05). The expression of p53,PCNA has significant difference in A₂,B₂ group at 1 week and 4 week(P<0. 05). Then there is no significant difference in C₂ group(P>0.05). The content of MDA, SOD and CAT has significant difference between the groupA₂ between 1 week and 4 week(P <0.05). However, there' s no significant difference in groupB₂. At 4 week, there' s no significant difference in three groups of SOD(P>0. 05). There' s significant difference between A₂ and C₂ of CAT(P<0. 05). (3) There were no the acute toxicity, irritation effect, when "CuYuBanHenXiao" was spreaded on the rabbits' normal and damaged skin and no sensitivity effect phenomenon to rabbits' nomal skin.Conclusion "CuYuBanHenXiao" can promote wound healing and prevent the hypertrophic scar of rabbit ears. It can encourage fibroblast of scar apoptosis and prevent fibroblast of scar proliferation. It can elevate the content of trace element of scar(Zn ,Cu ,Fe ,Mn) and provid the necessary of enzyme(SOD,CAT) to remove free radical. There were no the acute toxicity, irritation effect, when "CuYuBanHenXiao" was spreaded on the rabbits' normal and damaged skin and no sensitivity effect phenomenon to rabbits' nomal skin. So it' s safe to Applie to clinical.