论文部分内容阅读
目的对比观察小鼠放射性复合切割伤与单纯切割伤愈合过程中粒细胞-巨噬细胞集落刺激因子(granulocytemacrophage colony stimulating factor,GM-CSF)动态表达水平的差异以及GM-CSF对小鼠放射性复合切割伤的干预治疗效果,进一步探讨GM-CSF对放射性复合切割伤愈合的影响。方法 56只雌性昆明种小鼠(20~22g)随机分为辐照组和对照组,各28只,辐照组小鼠在6Gy 60 Coγ射线全身一次性均匀辐照后,即刻于背部皮肤制作全层缺损伤口,构建放射性复合切割伤模型;对照组小鼠伤口制备部位、方法同实验组,但小鼠不作辐照处理。伤后1、3、5、7d时分别处死小鼠7只,取创面周围皮肤及下方的薄层肌肉组织,通过Real-time PCR和免疫组化方法检测伤口愈合过程中GM-CSF的mRNA及蛋白表达水平。另将40只同品系、同体质量小鼠随机分为两组(实验组和对照组,各20只),均按上述相同方法构建放射性复合切割伤小鼠模型,于伤后0、1、3、4、5、7、9、11、14d时分别给予rhGM-CSF(6 000ng/mL)凝胶或空白凝胶涂抹,通过检测创面残余面积与胶原纤维水平,评价rhGM-CSF凝胶对放射性复合切割伤口的治疗效果。结果致伤后1~3d辐照组GM-CSF的mRNA及蛋白水平均低于对照组(P<0.05或P<0.01);5~7d时,对照组GM-CSF的mRNA及蛋白表达逐渐下降,而辐照组GM-CSF水平无明显改变。凝胶涂抹4~11d内,rhGM-CSF凝胶治疗组中小鼠皮肤伤口残余面积低于空白凝胶组(P<0.05或P<0.01);Masson染色显示:与空白凝胶组相比,rhGM-CSF凝胶治疗后放射性复合切割伤口中可见显著增生、广泛分布,排列紧密的胶原纤维。结论放射性复合切割伤口早期愈合过程中GM-CSF的mRNA及蛋白表达水平较正常伤口低,而持续给予6 000ng/mL rhGM-CSF刺激可明显加快放射性复合切割伤的愈合进程。
Objective To compare the dynamic expression of granulocyte macrophage colony stimulating factor (granulocyte macrophage colony stimulating factor, GM-CSF) and radioactive compound-cleavage (GM-CSF) in mice after radiofrequency ablation and simple cutting wound healing Injury intervention treatment effect, to further explore the GM-CSF on radiofrequency cutting wound healing. Methods Fifty-six female Kunming mice (20-22g) were randomly divided into irradiation group and control group, 28 mice in each group. The irradiated mice were irradiated with 6Gy 60Coγ ray all over the body once Full-thickness wounds were sacrificed to establish a radioactive composite cutting wound model. The control group was given the same wound preparation site as the experimental group, but the mice were not irradiated. Seven mice were killed at 1, 3, 5 and 7 days after injury respectively. The skin around the wound and the thin layer of muscle tissue were examined. Real-time PCR and immunohistochemistry were used to detect the mRNA and protein expression of GM-CSF during wound healing. Protein expression levels. Another 40 same-line, same-body weight mice were randomly divided into two groups (experimental group and control group, each 20), according to the same method as above to construct a radioactive composite cutting injury mouse model, 0,1,3 , Respectively. On the 4th, 5th, 7th, 9th, 11th and 14th day, rhGM-CSF (6 000 ng / mL) gel or blank gel was applied respectively. The remnant area of the wound and collagen fiber level were measured to evaluate the effects of rhGM- Therapeutic effect of composite cutting wound. Results The mRNA and protein levels of GM-CSF in irradiation group 1 ~ 3d were lower than that in control group (P <0.05 or P <0.01). At 5 ~ 7d, the mRNA and protein expression of GM-CSF in control group decreased , While the level of GM-CSF in irradiation group did not change significantly. The residual area of mice skin wounds in the rhGM-CSF gel treatment group was lower than that in the blank gel group (P <0.05 or P <0.01) within 4 to 11 days after gel smear treatment. Masson staining showed that compared with the blank gel group, rhGM -CSF gel treatment of radioactive incision in the wound visible hyperplasia, widely distributed, closely arranged collagen fibers. Conclusions The mRNA and protein expression of GM-CSF in early healing of radioactive combined wound healing is lower than that of normal wound. Continuous 6ng / mL rhGM-CSF stimulation can significantly accelerate the healing process of radioactive combined wound healing.