热休克蛋白47重组质粒对糖尿病大鼠皮肤创口愈合影响的实验研究

目的：初步研究热休克蛋白47（HSP47）重组质粒pTraeer—CMV—HSP47局部注射对糖尿病大鼠皮肤创口愈合的影响，探讨以HSP47质粒为摹础的基因治疗促进糖尿病皮肤创口愈合的可行性和有效性．方法：制备四氧嘧啶诱导的糖尿病大鼠皮肤创口愈合模型，创缘皮下局部注射HSP47重组质粒，观察其对皮肤创口愈合的影响，并用免疫组织化学、荧光定量RT—PCR、Western blotting检测创口愈合过程中HSP47和collagen Ⅰ的表达变化。结果：在糖尿病大鼠皮肤刨缘连续两次注射质粒后第3、6、9天．免疫组织化学、荧光定量RT-PCR和Western blotting检测证实，HSP47重组质粒组比对照组能显著增加创口中的HSP47和collagenⅠ的表达。结论：在糖尿病大鼠皮肤创口周围注射质粒pTracer—CMV—HSP47，能明显增强HSP47和coilagenⅠ的表达，有助于改善创口愈合，HSP47可能是促进糖尿病皮肤创口愈合的潜在治疗靶点。

Effects of HSP47 Recombinant Plasmid on the Skin Wound Healing Process in the Alloxan-induced Diabetic Rats

Objective: To investigate the effects of local injection of HSP47 recombinant plasmid on the skin wound healing process in alloxan-induced diabetic rats and assess the possibility and utility of gene therapy based on HSP47 plasmid. Methods: Diabetic rats with skin wound were randomly distributed into 3 groups. Group A wounds injection normal saline, group B injection pTracer-CMV, group C injection pTracer-CMV-HSP47. Immunohistochemistry, quantitative fluorescent RT-PCR, Western blotting techniques during 3, 6, 9 d after plasmid injection were used to evaluate the expression changes of HSP47 and collagen I. Results: After two consecutive local plasmid injections, the expression of HSP47 and collagen I in pTracer-CMV-HSP47 group was concomitantly strengthened and much intensify than those in A and B group. Conclusion: The expression of HSP47 and collagen I were considerably enhanced after pTracer-CMV-HSP47 local injection, which may help to improve the wound healing in the diabetic rats. The data suggests that HSP47 may possibly become a potential therapeutic target to promote diabetic wound healing in future.