论文部分内容阅读
[摘要]目的：制备外用VEGF165-胰岛素复合凝胶并观察其对糖尿病大鼠深Ⅱ度烫伤创面愈合的作用。方法：以新型凝胶卡波姆980为基质，分别加入VEGF165、胰岛素，分别配制成胰岛素凝胶、VEGF165凝胶、VEGF165-胰岛素复合凝胶和空白对照凝胶。观察各组凝胶性状，检测其pH值。75只SPF级雄性Wistar大鼠，体重190～290g，随机分为正常对照组、糖尿病对照组、胰岛素凝胶组、VEGF165凝胶组、VEGF165-胰岛素复合凝胶组，每组15只。各组大鼠禁食12h后，除正常对照组外，其余各组一次性腹腔注射链脲佐菌素（55 mg/kg）制备1型糖尿病模型，正常对照组大鼠注射相同剂量柠檬酸钠缓冲液。1型糖尿病大鼠造模成功后，采用恒温水浴箱80℃，制备深Ⅱ度烫伤模型。正常对照组、糖尿病对照组创面外敷空白凝胶基质，其余各组外敷对应凝胶制剂，每天换药1次。深Ⅱ度烫伤造模成功后观察各组大鼠存活情况，于3、7、11、15、21天时点观察糖尿病大鼠创面愈合情况并计算创面愈合率，在各时点随机处死每组3只大鼠取烫伤皮肤全层留取标本，行组织学切片及免疫组化染色观察各时点炎性细胞，微血管形成及胶原蛋白。结果：制备的各组凝胶透明，保湿性极佳，黏附性良好，便于涂抹及清洗，无组织毒性。各组大鼠无感染及死亡发生。3天后烫伤各组创面无明显缩小，7、11、15、21天时，VEGF165-胰岛素复合凝胶组创面愈合率最高，糖尿病组最低。VEGF165-胰岛素复合凝胶组愈合率与其余各组比较，差异均有统计学意义（P < 0.05）。病理切片观察示各时点VEGF165-胰岛素复合凝胶组肉芽组织形成及上皮化均优于其余各组。免疫组化染色示各组微血管数量、Ⅰ型胶原蛋白阳性率均于3天开始表达，且随时间阳性细胞增多明显；各时间点VEGF165-胰岛素复合凝胶组微血管密度及Ⅰ型胶原蛋白最高，糖尿病组最低，与其余各组比较以及VEGF165-胰岛素复合凝胶组、糖尿病组间比较差异均有统计学意义（P < 0.05）。 结论：糖尿病大鼠皮肤深Ⅱ度烫伤外用VEGF165-胰岛素复合凝胶对创面愈合有明显的促进作用
[关键词]VEGF；胰岛素；卡波姆凝胶；糖尿病大鼠；创面愈合
[中图分类号]R587.1 [文献标识码]A [文章编号]1008-6455（2014）21-1799-07
Abstract： Objective Preparation of topical gel VEGF165-insulin complex and observe its deep Ⅱ de-gree burn wound healing in diabetic rats role. Methods Carbomer with new gel matrix， were added to VEGF165， insulin， insulin were formulated into a gel，VEGF165 gel， VEGF165-insulin composite gel and control gel. Gel characters of each group， the pH value of the detection. 75 SPF male Wistar rats weighing 190～290g， were randomly divided into normal control group， diabetic control group， insulin gel group， VEGF165 gel group， VEGF165-insulin gel composite groups 15. In addition to the normal control group， the other groups of rats were fasted 12h after intraperitoneal injection of streptozotocin （55mg/kg） Preparation of type 1 diabetes model， normal control rats injected at the same dose fasting sodium citrate buffer fluid. Type 1 diabetes in rats after successful modeling， using heated water bath 80℃，the preparation of deep Ⅱ degree burn model. Normal control group， diabetic control group topical wound gel matrix blank， the rest of the group corresponding topical gel formulation， dressing once a day. Deep Ⅱ degree burns after a successful modeling survival of the rats was observed at 3，7，11，15，21d point of wound healing in diabetic rats and calculate the rate of wound healing in each group were sacrificed at each time point 3 rats take full thickness skin burns specimens were taken for histological sections and immunohistochemical staining of inflammatory cells at each time point， angiogenesis and collagen. Results Each group prepared a transparent gel， moisturizing excellent， good adhesion， easy painting and cleaning， no tissue toxicity. The rats without infection and death. 3d no after burn wounds in each group was significantly reduced， 7，11，15，21d when VEGF165-insulin compound gel group the highest rate of wound healing， diabetic group was the lowest， VEGF165-insulin compound gel group healing rate with the rest of the group， the difference were statistically significant （P <0.05）. Histopathological examination shows each time point VEGF165-insulin compound gel group granulation tissue formation and epithelialization were better than the rest of the group. Immunohistochemical staining showed that the number of microvessels in each group，Ⅰcollagen expression positive rates start at 3d， and the positive cells was significantly increased with time； each time point VEGF165-insulin highest composite gel group microvessel density and collagen typeⅠdiabetes lowest group， with other groups and VEGF165-insulin compound gel group， the difference was statistically significant （P <0.05） between the diabetic group. Conclusion Diabetic rat skin deep Ⅱ degree burns topical gel VEGF165-insulin complex on wound healing have significant role in promoting
Key words：VEGF；insulin；carbomer gel；diabetic rats；wound healing
[关键词]VEGF；胰岛素；卡波姆凝胶；糖尿病大鼠；创面愈合
[中图分类号]R587.1 [文献标识码]A [文章编号]1008-6455（2014）21-1799-07
Abstract： Objective Preparation of topical gel VEGF165-insulin complex and observe its deep Ⅱ de-gree burn wound healing in diabetic rats role. Methods Carbomer with new gel matrix， were added to VEGF165， insulin， insulin were formulated into a gel，VEGF165 gel， VEGF165-insulin composite gel and control gel. Gel characters of each group， the pH value of the detection. 75 SPF male Wistar rats weighing 190～290g， were randomly divided into normal control group， diabetic control group， insulin gel group， VEGF165 gel group， VEGF165-insulin gel composite groups 15. In addition to the normal control group， the other groups of rats were fasted 12h after intraperitoneal injection of streptozotocin （55mg/kg） Preparation of type 1 diabetes model， normal control rats injected at the same dose fasting sodium citrate buffer fluid. Type 1 diabetes in rats after successful modeling， using heated water bath 80℃，the preparation of deep Ⅱ degree burn model. Normal control group， diabetic control group topical wound gel matrix blank， the rest of the group corresponding topical gel formulation， dressing once a day. Deep Ⅱ degree burns after a successful modeling survival of the rats was observed at 3，7，11，15，21d point of wound healing in diabetic rats and calculate the rate of wound healing in each group were sacrificed at each time point 3 rats take full thickness skin burns specimens were taken for histological sections and immunohistochemical staining of inflammatory cells at each time point， angiogenesis and collagen. Results Each group prepared a transparent gel， moisturizing excellent， good adhesion， easy painting and cleaning， no tissue toxicity. The rats without infection and death. 3d no after burn wounds in each group was significantly reduced， 7，11，15，21d when VEGF165-insulin compound gel group the highest rate of wound healing， diabetic group was the lowest， VEGF165-insulin compound gel group healing rate with the rest of the group， the difference were statistically significant （P <0.05）. Histopathological examination shows each time point VEGF165-insulin compound gel group granulation tissue formation and epithelialization were better than the rest of the group. Immunohistochemical staining showed that the number of microvessels in each group，Ⅰcollagen expression positive rates start at 3d， and the positive cells was significantly increased with time； each time point VEGF165-insulin highest composite gel group microvessel density and collagen typeⅠdiabetes lowest group， with other groups and VEGF165-insulin compound gel group， the difference was statistically significant （P <0.05） between the diabetic group. Conclusion Diabetic rat skin deep Ⅱ degree burns topical gel VEGF165-insulin complex on wound healing have significant role in promoting
Key words：VEGF；insulin；carbomer gel；diabetic rats；wound healing