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《The Journal of Traditional Chinese Orthopedics and Traumatology》 2015-11
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Impact of Taohong Siwu Tang(桃红四物汤) on Delta-like 4 expression in the autologous skin graft model of rabbits

LIN Nianci;SUN Shaoqiu;LI Yiliang;XU Wuji;CHEN Xilong;ZHOU Zhaohui;Traditional Chinese Medicine Hospital of Xinhui district in Jiangmen City;The Second Affiliated Hospital of Hunan University of Traditional Chinese Medicine;  
Objective: To observe the impact of Taohong Siwu Tang( 桃红四物汤,THSWT) on Delta-like 4 expression in the autologous skin graft model of rabbits. Methods: Seventy-five New Zealand rabbits which ranged in age from 10 to 12 months were randomly divided into sham-operated group,model group,low-dose group,middle-dose group and high-dose group,15 cases in each group. The autologous skin transplantation surgery were performed on rabbits in model group,low-dose group,middle-dose group and high-dose group,while sham operation were performed on rabbits in sham-operated group. After the modeling operation,the rabbits in sham-operated group and model group were intragastric administrated with distilled water; while the others in low-dose group,middle-dose group and high-dose group were intragastric administrated with the prepared THSWT in dosages of 2. 5,5 and 10 g / kg at a time,twice a day. Five rabbits were randomly selected from each group and the blood samples were obtained from the heart at 2,5 and 8 days after the beginning of the drug intervention respectively. The rabbits were executed and their anastomotic skin graft tissues were fetched out. The serum content of Delta-like 4 protein were measured by using enzyme-linked immunosorbent assay( ELISA). The acquired skin tissues were divided into 2 parts,and one of them were maded into paraffin sections and the blood vessel density were measured after HE staining,while the other portion were applied to measure the Delta-like 4 mRNA level by using reverse transcription-polymerase chain reaction( RT-PCR). Results: There was no statistical difference in serum content of Delta-like 4 protein between the 5 groups at 2 days after the beginning of the drug intervention( 30. 58 + /-3. 56,32. 31 + /- 2. 66,33. 53 + /- 1. 75,36. 00 + /- 1. 49,35. 63 + /- 1. 87 ng / ml,F = 2. 669,P = 0. 062). There was statistical difference in serum content of Delta-like 4 protein between the 5 groups at 5 and 8 days after the beginning of the drug intervention( 30. 56 + /- 1. 35,33. 52 + /- 2. 50,37. 96 + /- 1. 83,48. 74 + /- 2. 34,49. 05 + /- 2. 86 ng / ml,F = 46. 239,P = 0. 000; 29. 26 + /- 2. 20,32. 98 + /- 1. 43,35. 24 + /- 1. 97,38. 02 + /- 1. 72,36. 06 + /- 2. 50 ng / ml,F = 8. 394,P = 0. 000). There was no statistical difference in serum content of Delta-like 4 protein between model group and sham-operated group and between model group and low-dose group( 5d: P = 0. 112,P =0. 269; 8d: P = 0. 340,P = 0. 182). The serum content of Delta-like 4 protein were higher in middle-dose group and high-dose group compared to model group( 5d: P = 0. 000,P = 0. 000; 8d: P = 0. 020,P = 0. 006). There was no statistical difference in serum content of Deltalike 4 protein between middle-dose group and high-dose group( P = 0. 862,P = 0. 245). The serum content of Delta-like 4 protein were higher in middle-dose group and high-dose group compared to low-dose group at 5 days after the beginning of the drug intervention( P =0. 000,P = 0. 000). There was no statistical difference in serum content of Delta-like 4 protein between middle-dose group and low-dose group and between high-dose group and low-dose group at 8 days after the beginning of the drug intervention( P = 0. 105,P = 0. 623). There was statistical difference in mRNA level of Delta-like 4 in skin graft between the 5 groups at 2,5 and 8 days after the beginning of the drug intervention( 0. 26 + /- 0. 03,0. 29 + /- 0. 04,0. 34 + /- 0. 06,0. 42 + /- 0. 03,0. 39 + /- 0. 04,F = 7. 072,P = 0. 001; 0. 25 + /- 0. 03,0. 34 + /- 0. 04,0. 39 + /- 0. 03,0. 48 + /- 0. 06,0. 45 + /- 0. 06,F = 12. 028,P = 0. 001; 0. 25 + /- 0. 06,0. 30 + /- 0. 04,0. 35 + /- 0. 03,0. 44 + /- 0. 03,0. 43 + /- 0. 04,F = 10. 813,P = 0. 000). There was no statistical difference in Delta-like 4 mRNA level between model group and sham-operated group and between model group and low-dose group( 2 d: P = 0. 452,P = 0. 162; 5 d: P = 0. 320,P = 0. 158; 8 d:P = 0. 171,P = 0. 219). The Delta-like 4 mRNA level were lower in model group compared to middle-dose group and high-dose group( 2 d:P = 0. 000,P = 0. 001; 5 d: P = 0. 001,P = 0. 009; 8 d: P = 0. 001,P = 0. 002). There was no statistical difference in Delta-like 4 mRNA level between middle-dose group and high-dose group( P = 0. 426,P = 0. 330,P = 0. 725). There was no statistical difference in Delta-like 4mRNA level between middle-dose group and low-dose group and between high-dose group and low-dose group at 2 days after the beginning of the drug intervention( P = 0. 111,P = 0. 167). The Delta-like 4 mRNA level was higher in middle-dose group and high-dose group compared to low-dose group at 5 and 8 days after the beginning of the drug intervention( 5 d: P = 0. 024,P = 0. 025; 8 d: P = 0. 013,P =0. 028). There was statistical difference in the number of blood vessels in skin graft under the optical microscope( × 400) between the 5groups at 2 and 5 days after the beginning of the drug intervention( 5. 37 + /- 0. 36,2. 55 + /- 0. 41,2. 83 + /- 0. 24,3. 31 + /- 0. 26,3. 53 + /- 0. 67,F = 4. 084,P = 0. 014; 5. 62 + /- 0. 56,4. 25 + /- 0. 68,4. 80 + /- 0. 74,5. 88 + /- 0. 42,5. 92 + /- 0. 48,F = 4. 600,P =0. 009). The number of blood vessels was less in model group compared to sham-operated group( P = 0. 001,P = 0. 011),and there was no statistical difference in the number of blood vessels between model group and low-dose group( P = 0. 527,P = 0. 268) and between middledose group and high-dose group( P = 0. 620,P = 0. 939). At 2 days after the beginning of the drug intervention,there was no statistical difference in the number of blood vessels between middle-dose group and model group( P = 0. 102) and between high-dose group and model group( P = 0. 131) and between middle-dose group and low-dose group( P = 0. 297) and between high-dose group and low-dose group( P =0. 352). At 5 days after the beginning of the drug intervention,the number of blood vessels was more in middle-dose group and high-dose group compared to model group( P = 0. 003,P = 0. 003),while no statistical difference was found between middle-dose group and low-dose group and between high-dose group and low-dose group( P = 0. 038,P = 0. 033). At 8 days after the beginning of the drug intervention,there was no statistical difference in the number of blood vessels in skin graft under the optical microscope( × 400) between the 5 groups( 6. 18 +/- 0. 33,5. 86 +/- 0. 66,6. 03 +/- 0. 40,6. 27 +/- 0. 53,6. 27 +/- 0. 59,F = 0. 334,P = 0. 852). Conclusion: THSWT can up-regulate Delta-like 4 expression and promote angiogenesis in autologous skin graft model of rabbits. However,we cann't clarify the relationship between the role of THSWT in up-regulating Delta-like 4 expression and its dose and action time,which need to be further studied.
【Fund】: 国家自然科学基金项目(81373658)
【CateGory Index】: R285.5;R-332
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