真皮多能干细胞移植治疗大鼠皮肤光老化的实验研究

发布时间：2018-06-19 20:08

本文选题：真皮多能干细胞 + 皮肤光老化　；参考：《遵义医学院》2010年硕士论文

【摘要】： 目的：本研究拟探讨真皮多能干细胞(dermal multipotential stem cells, DMSCs)移植对大鼠皮肤光老化的修复作用,为皮肤光老化提供新的治疗方法。方法：57只SD大鼠随机分为正常组8只和模型组49,采用UVA和UVB紫外灯同时对模型组进行照射(每天2h,连续照射60天),随机取8只大鼠进行皮肤光老化模型鉴定,模型成功后随机分为对照组16只(PBS代替)和治疗组25只(1×106细胞悬液)。体外分离培养SD乳鼠DMSCs,经鉴定后用DAPI标记,取治疗组大鼠9只,皮肤光老化区多点真皮层内移植细胞悬液,分别在移植后1d、3d、7d处死大鼠各3只,在荧光显微镜下观察DAPI标记的DMSCs在皮肤光老化区域组织存活、迁徙和定居。然后对剩余治疗组大鼠皮肤光老化区进行细胞移植；4周、8周分别采用免疫组织化学(波形蛋白)、VG和Weigert染色方法检测各组真皮中成纤维细胞、胶原纤维和弹力纤维的变化。结果：在不同时间段检测到DAPI标记的DMSCs在皮肤光老化区存活、扩散、迁移。免疫组织化学染色结果显示：与对照组相比：4周后治疗组成纤维细胞增多,主要在真皮浅层较多,两者比较其差异有统计学意义(P0.01),8周后广泛均匀弥散在皮肤各层,两者比较其差异有统计学意义(P0.01)。VG染色结果显示：与对照组相比：4周后治疗组变性、成块胶原纤维含量减少,可见少量新生胶原纤维,8周后延伸至皮下组织及真皮深层,变性胶原纤维基本消失,出现大量染色深、排列有序的新生胶原纤维,皮肤结构层次清晰,与正常组无明显变化。Weigert染色结果显示：与对照组相比：4周后治疗组真皮深层聚集、缠结的变性弹力纤维明显减少,8周后真皮浅层变性弹力纤维基本消失,毛囊周围及真皮浅层见纤细新生波浪状弹力纤维,与正常组无明显变化。结论：DMSCs移植对大鼠皮肤光老化有明显的治疗作用。
[Abstract]:Objective: to investigate the effect of dermal pluripotent stem cell transplantation on photoaging of rat skin, and to provide a new therapeutic method for skin photoaging. Methods Fifty-seven Sprague-Dawley rats were randomly divided into normal group (n = 8) and model group (n = 49). The model group was irradiated with UVA and UVB ultraviolet lamp at the same time. After success, the model was randomly divided into control group (n = 16) and treatment group (n = 25). SD neonatal rats were isolated and cultured in vitro. After identification, 9 rats in the treatment group were labeled with DAPI, and 3 rats in each group were killed at 1 day after transplantation, 3 rats were killed on the 3rd day after transplantation. DAPI labeled DMSCs were observed to survive, migrate and settle in skin photoaging area under fluorescence microscope. The changes of fibroblasts, collagen fibers and elastic fibers in the dermis of the remaining treatment group were detected by immunohistochemical staining (vimentin VG and Weigert staining) after 4 weeks and 8 weeks of cell transplantation. Results: DAPI labeled DMSCs were detected to survive, diffuse and migrate in skin photoaging area at different time points. The results of immunohistochemical staining showed that compared with the control group, the number of fibroblasts increased after 4 weeks of treatment, mainly in the superficial dermis. The difference between the two groups was statistically significant after 8 weeks. The difference between the two groups was statistically significant (P 0.01). VG staining showed that compared with the control group, the denaturation of collagen fibers in the treatment group decreased after 4 weeks compared with the control group. It was found that a small amount of new collagen fibers extended to the subcutaneous tissue and the deep dermis after 8 weeks. The denatured collagen fibers disappeared, a large number of new collagen fibers were stained deeply, the skin structure was clear and the skin structure was clear. The results of Weigert staining showed that: compared with the control group, the treatment group had deep dermal aggregation after 4 weeks. The denatured elastic fibers of tangles decreased significantly after 8 weeks, the denatured elastic fibers in the superficial dermis disappeared basically, and the new wave elastic fibers were found around the hair follicles and in the superficial layers of the dermis, which had no obvious change from the normal group. Conclusion\
【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R758.1

【参考文献】