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Objective: To observe the bitter soup topical herbal compound denitrification on the rabbit back Renfen contaminated surgical wound healing index, the number of fibroblasts, capillaries, TGF-β1 protein content and EGF protein content, to verify denitrification bitter soup topical promote hemorrhoid surgery The model of wound healing and its role in the prevention of excessive scar wound investigate the saltpetre bitterness soup to promote wound healing and the mechanism of inhibition of wound excessive scarring. Method: 1. Wound model making grouping: 16 New Zealand white rabbits, male and female, electronic weighing scales, the rabbit both sides back with 8% sodium sulfide hair removal, and then 25% urethane (4ml/kg) anesthesia . Back central spinal anesthesia in the rabbit after the success of each side of the next open 1.5cm from front to back, each cutting two circular full-thickness skin wounds, diameter 1.8cm, area 1.54cm2. To add the liquid manure 1ml (fresh human feces 10g, plus saline 5 ml of mixing, centrifuged, and the supernatant was now with the current) in the wound after the full stop bleeding, gauze, dressings, adhesive tape, was observed after 48 hours wounds, according to the literature, see the wound purulent secretions and fecal odor is the successful modeling. 64 wounds of the 16 model rabbits were randomly divided into four groups (n = 16 wounds): A set of blank control group, group B rhEGF treatment group, C group for Chinese medicine denitrification bitter decoction group, D group for comprehensive treatment group (traditional Chinese medicine rhEGF). Administration: the day from the success of the model began administration. Group A: daily Tu saline once again dressings, tape fixed dressing was changed daily until the wounds healed. Group B: uniform spraying of rhEGF (120AU/cm2) inferior to the wounds, I with the A group. C: Tu the denitrification bitter soup 40mg/cm2, I with the A group. Group D: the uniform spraying rhGF (120AU/cm2), 30 minutes after coating the denitrification bitter soup (40mg/cm2), the same group A These groups before each dose half an hour in the wound coated fecal 1ml daily medication before routine cleaning wounds, dressing the same time. Drawn: 4,6,8,10,12,14,16,18 days after medication measuring the wound area, calculate the healing index, and in each group, respectively, at the same number of days to take a total of eight animals, anesthesia ( were killed after the success of the urethane 4ml/kg), wound granulation tissue and isolated ophthalmic surgical scissors into blocks remove specimens with 10% neutral formalin fixed after 24 hours, with 75% alcohol to soak for 24 hours then the specimen was soaked into 70% alcohol to be examined (conducted to the number of fibroblasts, and counting the number of capillary, detection) of the EGF protein. The remaining eight animals in the healing 1,2,3,4,5,6,7,8 days were sacrificed an animal wound scar tissue isolated ophthalmic surgical scissors, and into the block remove, with the use of the law deal with After waiting for inspection (detection of TGF-β1 protein expression). Detection: Remove the specimens, embedded in paraffin, respectively HE staining, and then compared the pathological examination (number of fibroblasts, capillaries); simultaneous use SABC immunohistochemical staining (same kit), respectively immunohistochemistry slice of TGF-β1, EGF protein expression detected IPP image analysis software analysis, results are the average gray value. OUTCOME MEASURES: ① the wound healing Index (Healing index); the ② wound granulation tissue fibroblasts Quantity (Fibroblasts number); ③ granulation tissue capillaries (Capillary number); ④ EGF protein content of granulation tissue (f epidermal cell factor); ⑤ wounds scar tissue expression of TGF-β1 protein content (Transforming growth factor). 6. Statistical Methods: The experimental data obtained are x ± s, t-test for differences between groups, P lt; 0.05 was considered statistically significant difference. Results: 1. Affect wound healing Index: ① The the denitrification bitter soup group average wound healing index with the saline group (t = 2.97, P lt; 0.05), the difference was significant, statistically significant; and comprehensive treatment group (t = 2.19, P lt; 0.05), the difference was statistically significant; rhEGF group (t = 0.09, P gt; 0.05), the difference was not significant and was not statistically significant. ② saline group compared with rhEGF group (t = 2.66, P lt; 0.05), the difference was significant, statistically significant; compared with the combined therapy group (t = 4.36, P lt; 0.05), the difference was statistically significant . Comparison the ③ rhEGF group and the treatment group (t = 2.20 P lt; 0.05), the difference was significant, with statistical significance. Number of fibroblasts: ① The the denitrification bitter soup group wound the average number of fibroblasts with the saline group comparison, the difference was significant, statistically significant (t = 17.47, P lt; 0.001); with the Comprehensive treatment group, the difference was significant, statistically significant (t = 2.67, P lt; 0.05); comparison with rhEGF group, the difference was not statistically significant, was not statistically significant (t = 0.26, P gt; 0.05). ② saline group compared with rhEGF group, significant differences was statistically significant (t = 18.06, P lt; 0.001); compared with the combined therapy group, the difference was significant, statistically significant (t = 20.86, P lt; 0.05). ③ rhEGF groups compared with the combined therapy group, the difference was significant, statistically significant (t = 2.32, P lt; 0.05). Impact on the number of capillaries: the average number of capillaries ① The the denitrification bitter soup group wound with saline group, the difference was significant, statistically significant (t = 7.84, P lt; 0.001); comprehensive treatment group significant difference was statistically significant (t = 2.42, P lt; 0.05); comparison with rhEGF group, the difference was not significant and was not statistically significant (t = 0.29, P gt; 0.05). ② saline group compared with rhEGF group, significant differences was statistically significant (t = 7.74, P lt; 0.001); compared with the combined therapy group, the difference was significant, statistically significant (t = 10.37, P lt; 0.001); ③ of rhEGF group with combined therapy group, the difference was significant, statistically significant (t = 2.75, P lt; 0.05). 4. Groups granulation tissue EGF protein (gray value) changes: ① bitter decoction group and the saline group, the difference was significant, statistically significant (t = 7.20, P lt; 0.001); and comprehensive treatment significant group difference was statistically significant (t = 2.34, P lt; 0.05); comparison with rhEGF group, the difference was not statistically significant, was not statistically significant (t = 0.04, P gt; 0.05). ② saline group compared with rhEGF group, significant differences was statistically significant (t = 7.38, P lt; 0.001); compared with the combined therapy group, the difference was significant, statistically significant (t = 13.03, P lt; 0.001). ③ rhEGF groups compared with the combined therapy group, the difference was significant, statistically significant (t = 2.36, P lt; 0.05). Of TGF-β1 protein expression (gray value): ① The the denitrification bitter soup group comparison with the saline group, the difference was significant, statistically significant (t = 7.30, P lt; 0.001); and comprehensive treatment significant group difference was statistically significant (t = 5.48, P lt; 0.001); comparison with rhEGF group, the difference was not significant, no statistical significance (t = 10.29, P lt; 0.001). ② saline group compared with rhEGF group, significant differences was statistically significant (t = 2.98, P lt; 0.001); compared with the combined therapy group, the difference was significant, statistically significant (t = 0.29, P gt; 0.05). ③ rhEGF groups compared with the combined therapy group, the difference was significant, statistically significant (t = 2.76, P lt; 0.05). Conclusion: This study showed that denitrification the bitter soup can promote the rabbit back Renfen pollution surgical wound healing; 2 This experiment confirmed the denitrification bitter soup able to prevention the rabbit back Renfen contaminated surgical wound healing, excessive scarring; 3. Experimental results show that the denitrification bitter soup promote wound healing may promote wound EGF and TGF-β1 expression; 4. saltpeter the bitter soup suppression wound excessive scarring speculate that the inhibition of the scar tissue TGF-β1 expression; 5. studies suggest that nitric bitter soup joint application and rhEGF better promote the healing of the surgical wound the rabbit back Renfen pollution; 6 The results also suggest Chinese medicine topical can promote the hemorrhoid surgery wound healing and prevent wound excessive scarring, and through a variety of mechanisms play a therapeutic role.
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