胶原酶软膏修复大鼠深Ⅱ度烫伤创面及其对表皮生长因子、成纤维细胞生长因子表达的影响

Effect of collagenase ointment on expression of epidermal growth factor, basic fibroblast growth factor and wound repair in rats with partial-thickness burn

目的观察胶原酶软膏局部用药对大鼠背部深Ⅱ度烫伤创面的修复作用及其对表皮生长因子(EGF)、成纤维细胞生长因子(bFGF)表达的影响。方法将20只雄性SD大鼠编号后随机分为4组,每组5只,每只大鼠背部造两个深Ⅱ度烫伤创面,随机选取一个创面作为实验组,另一个创面作为对照组。实验组用胶原酶软膏换药,对照组用凡士林纱布换药,1次/d,直至创面愈合。每日观察大鼠创面分泌物、痂皮形成与分离程度,记录愈合时间,定期测量愈合面积,计算愈合率。自伤后7d、10d、14d、21d各取已分编组的4组中的一组大鼠自创缘取材做病理及EGF、bFGF免疫组化切片,免疫组化切片高倍镜下人工计数阳性细胞数。实验数据以x珋±s表示,采用SPSS17.0统计软件进行处理,组间比较采用配对t检验。结果实验组创面愈合时间(20.75±1.71)d明显短于对照组(25.25±1.71)d(t=6.971,P=0.006)。7d、10d、14d、21d愈合率分别为(37.60±7.42)%、(54.52±7.20)%、(77.72±8.50)%、(98.04±3.92)%,均高于对照组(19.06±4.26)%、(35.96±5.88)%、(49.48±6.04)%、(81.80±6.04)%(t值分别为8.200、9.116、10.102、5.294,P值均小于0.05)。病理切片显示实验组创面愈合后瘢痕形成较实验组轻,镜下可见胶原蛋白细小、排列有序;实验组皮肤附件结构恢复好,皮下可见毛囊形成。免疫组化切片显示实验组7d、14dEGF阳性表达均高于对照组(t值分别为2.671,3.127;P值分别为0.022,0.010),10d明显高于对照组(t=6.585,P<0.001),21d表达与对照组比较差异不具有统计学意义(t=0.309,P=0.763)。bFGF7d、14d阳性表达均高于对照组(t值分别为2.370、2.918;P值分别为0.037、0.014),10d明显高于对照组(t=9.106,P<0.001),21d表达与对照组比较差异不具有统计学意义(t=0.132,P=0.898)。结论胶原酶软膏对深Ⅱ度创面具有明显的促进愈合作用,其机制与加速坏死组织分离、减轻局部炎性反应、增加EGF和bFGF的表达有关。

Objective To obserae the effect of collagenase ointment on expression of epidermalgrowth factor（EGF）, basic fibroblast growth factor （bFGF）and wound repair in rats with partial-thickness burn. Methods 20 male SD rats were numbered, made two wounds in the back, and randomly divided into experiment group, and control group. The experiment group was treated with collagenase ointment, the control group with vaseline gauze, once a day, until wound healing. The wound exudates, crusta formation and separating degree were observed every day. The healing time was recorded, the healing area was measured periodically to calculate the rate of healing. Tissues of one group with 5 rats, were collected from the edge of wound in 7, 10, 14, 21 d. The tissue was made into pathological and EGF, bFGF immunohistochemical sections. The number of positive cells was counted under high power microscope. The experimental data were expressed as mean ± standard deviation, using SPSS17. 0 statistical software, Comparision between groups was made using the paired t test. Results The wound healing time of experiment group was （20.75 ± 1.71 ） d, significantly shorter than that in the control group （25.25 ± 1.71 ） d （ t = 6. 971, P = 0. 006）. The 7, 10, 14, 21 d healing rates of experiment group were （37.60 ± 7.42 ） %, （ 54.52 ± 7.20 ） %, （ 77.72 ± 8.50） %, （ 98.04 ± 3.92 ） %, higher than that of control group [（19.06±4.26）%,（35.96±5.88）%, （49.48 ±6.04）%, （81.80 ±6.04）%] （t = 8. 200, 9. 116, 10. 102, 5. 294 ; P 〈 0.05）. Pathological examination showed that the experimental group was better than the control group in the quality of wound healing, hnmunohistochemistry revealed that the positive expression of EGF in the experimental group was higher than that of control group （t = 2. 671, 3. 127; P = 0. 022, 0.010） in 7, 14 d, and significantly higher than the control group （P = 6. 585, P 〈 0. 001 ） in 10 d. Difference of the expression between experimental group and control group had statistical significance （ t = 0. 309, P = 0. 763）. The positive expression of bFGF in the experimental group were higher than that of control group （t = 2. 370, 2. 918 ; P = 0. 037, 0. 014） in 7, 14 d, and significantly higher than the control group （P = 9. 106, P 〈 0. 001 ） in 10 d. Difference of the expression between experimental group and control group had statistical significance （t = 0. 132, P = 0. 898 ）. Conclusion Collagenase ointment promotes wound repair in rats with partial-thickness burn. It mechanism was related to that collagenase can accelerate the separation of necrotic tissue, reduce local inflammatry, response, increased the expression of EGF and bFGF in wound.