摘要
目的观察骨髓间充质干细胞(BMSCs)在治疗SD大鼠糖尿病溃疡创面愈合中成纤维细胞的变化,揭示BMSCs促进创面愈合的机制。方法提取同种异体SD大鼠BMSCs,传至第3、4代备用。将SD大鼠分为3组,空白对照组(A组)、糖尿病对照组(B组)、BMSCs治疗组(C组),每组20只。A组普通饲料喂养,B组、C组采用高脂高糖饮食+小剂量链脲佐菌素注射建立糖尿病大鼠模型,成模后,在SD大鼠背部取直径为2cm创面,3d后,溃疡模型成模。B组取密度为1×109/L的BMSCs1mL的DMEM-F12培养基悬浊液注射于创面,A组、C组取1mL0.9%氯化钠溶液注射于创面,分别于即刻、3d、7d、14d、21d取创面组织制备切片,苏木精-伊红(HE)染色,观察成纤维细胞生长情况,并提取组织中成纤维细胞,观察其增殖活力以及创面愈合情况。结果高糖高脂饮食+小剂量链脲佐菌素注射1周后测血糖,A组大鼠血糖为(3.91±0.44)mmol/L,B组血糖为(21.29±0.45)mmol/L,C组血糖为(21.46±0.51)mmol/L,B组、C组血糖明显高于A组(P<0.05)。BMSCs治疗7d,计算创面愈合率,A组(26.91±2.12)%,B组(7.55±2.11)%,C组(20.64±2.54)%,与A组和C组相比,B组愈合率明显降低(P<0.01)。组织病理可见,与B组相比,C组创面组织中成纤维细胞数量明显增多,细胞核饱满,成纤维细胞增殖活力较强;C组创面组织成纤维细胞增殖率较A组低,但高于B组。结论在糖尿病溃疡创面愈合过程中,局部注射同种异体BMSCs可促进糖尿病溃疡创面局部成纤维细胞生长,从而促进创面愈合。
Objective To observe the change of fibroblasts in diabetes ulcer wound healing with the treatment of bone marrow mesenehymal stem cells （ BMSCs）, thus to elaboration the mechanism of BMSCs promoting wound healing. Methods BMSCs were isolated from the bone marrow of rats, then purified and cultured, and the third or fourth passage of the BMSCs were spared. All of the SD rats were divided into group A （blank control group）, group B （diabetes control group） and group C （BMSCs treatment group）. Every group includes twenty rats. The rats in group A were used ordinary feed. The rats in group B and group C were used high fat and sugar diet and injected small doses of streptozocin to establish diabetes model. Then rats were took a wound that the diameter was 2 centimeters. Three days later, the ulcer model was completed. BMSCs at a density of 1 × 109/L in 1 milliliter DMEM/F12 were injected into the wounds of the rats in group C. The rats in group A and group B were injected with sodium chlorides. Then the wound tissue sections were prepared at once, after 3 days, 7 days, 14 days and 21 days. After Hematoxylin-eosin （HE） staining, the fibroblast growth situation was observed. In the meantime, fibroblasts were picked up and observed the proliferation activity. Results The blood glucose was easured after one week that rats were plused small doses of streptozotocin injection. The blood glucose in group A was （3.91 ± 0.44） mmol/L, group B was （21.29 ± 0.45） mmol/L, and group C was （21.46± 0.51） mmol/L. The results in group B and group C were higher than that in group A （P 〈 0.05 ）. At the seven days after the treatment, the rates of wound healing were calculated. Group A was （ 26.91 ± 2.12 ） %, group B was （ 7.55 ±2.11 ） %, and group C was （20.64± 2.54） %. The histopathology showed fibroblasts increased significantly in the wound tissue of group C than that of group B. The fibroblasts nucleus were full and fibroblast proliferated activity. MTT experiment confirmed that the rate of fibroblast proliferation in the wound tissue of group C was lower than that of group A, but higher than that of group B. Conclusions During diabetic wound healing, BMSCs could promoted diabetes ulcer wound healing by promoting the growth of local fibroblasts.
出处
《中华损伤与修复杂志（电子版）》
CAS
2013年第4期13-17,共5页
Chinese Journal of Injury Repair and Wound Healing(Electronic Edition)
基金
北京市自然科学基金(7122179)
全军医学科技"十二五"重点课题(BWS11C061)
