| 引用本文: | 梁至洁,黄东琳,朱丹丹,易晓林,宁艳,蒋洪棉,罗丽凤.淫羊藿苷优化脂肪干细胞促进大鼠超长随意皮瓣成活的实验研究[J].中国临床新医学,0,():-. |
| LIANG Zhi-jie.淫羊藿苷优化脂肪干细胞促进大鼠超长随意皮瓣成活的实验研究[J].中国临床新医学,0,():-. |
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| 摘要: |
| 目的 探讨淫羊藿苷(ICA)优化大鼠脂肪干细胞(rASCs)促进超长随意皮瓣存活的作用及机制。方法 提取rASCs并鉴定其多向分化能力及表面标记物。通过蛋白免疫印迹（WB）检测不同浓度ICA（0-10μM）对rASCs中血管生成因子（VEGF、PDGF-BB、FGF-2、IGF-2）蛋白表达的影响。构建大鼠超长随意皮瓣模型，根据干预方式设对照组，rASCs组及ICA+rASCs组。术后7d、14d及28d评估皮瓣存活率；28d取皮瓣组织行组织学及WB试验分别检测组织中微血管密度（MVD）、胶原含量及各血管生成因子蛋白的表达。结果 rASCs具有成脂、成骨和软骨分化的能力，CD29及CD44呈高表达，不表达CD45。WB结果提示1及10μM的ICA刺激能显著提高rASCs中血管生成因子的表达（P<0.05）。动物实验提示与对照组相比，在术后7d、14d，rASCs组及ICA+rASCs组的大鼠皮瓣成活率明显提高(P<0.05)；在术后28d，仅有ICA+rASCs组的皮瓣成活率显著高于对照组 (P<0.05)。皮瓣的组织学及WB结果显示，与对照组相比，rASCs组和ICA+rASCs组的皮瓣组织结构排列更为紧密，MVD、胶原含量及各管生成因子蛋白的表达显著提高（P<0.05）；此外，相对于rASCs组，ICA+rASCs组的MVD及胶原含量更高（P<0.05），VEGF及PDGF-BB的表达水平也显著高于rASCs组（P<0.05）。结论 ICA能够提高rASCs血管生成因子的表达水平，这可能是其优化rASCs促进大鼠超长随意皮瓣存活的机制。 |
| 关键词: 脂肪来源干细胞 淫羊藿苷 超长随意皮瓣 血管新生 |
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| 基金项目:国家自然科学基金资助项目(编号81760346,81860341)；南宁市优秀青年科技创新创业人才培育项目(编号RC20180201)；广西自然科学(编号2018GXNSFAA281148) |
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| Experimental study of icariin optimizing adipose-derived stem cells to promote the survival of ultra-long random skin flaps in rats |
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LIANG Zhi-jie1,2,3
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1.The First People'2.'3.s Hospital of Nanning
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| Abstract: |
| Objective To explore the effect and mechanisms of Icarin (ICA) to optimize rat adipose stem cells (rASCs) in promoting the survival of ultra-long random skin flaps. Methods To isolate and culture rASCs, and identify their surface factors' expression and multilineage differentiation ability. Western Blot (WB) was used to assess the protein expression of angiogenesis factors (VEGF、PDGF-BB、FGF-2、IGF-2) in rASCs stimulated by ICA at different concentrations (0-10μM). To establish rat models of ultralong random skin flap, and to inject PBS (Ctrl group), rASCs (rASCs group), and rASCs treated with 1μM ICA (ICA+rASCs group) into the pedicle of the flaps respectively. Then evaluate the surcival rate of flaps at 7d、14d and 28d after the operation. After 28d the survivor tissue of flap was dissected, to compare the microvessel density (MVD) and collagen content (%) in each group by histological detection. And the protein expression of angiogenesis factors in flap tissue is detected by WB. Results rASCs had the ability of adipogenic, osteogenesis, and cartilage differentiation. Flow cytometry experiments showed that rASCs had high expression of CD29 and CD44, and low expression of CD45. WB test suggested that 1μM and 10μM ICA could significantly improve the expression of angiogenesis factors in rASCs. Animal experiments showed that at 7d and 14d after the operation, compared with the control group, rASCs and ICA pretreated rASCs could improve flap survival rate (P<0.05), but there was no significant difference between these two groups. However, there was no significant difference in flap survival rate between rASCs group and the control group, while there was still a statistical difference between ICA+rASCs group and the control group (P<0.05) at 28d. HE staining showed that compared with the control group, the dermal fibers were more compact in the rASCs group and the ICA+rASCs group. The collagen content, MVD, and the expression of angiogenesis factors in the flap tissue were significantly higher in rASCs group and the ICA+rASCs group (P<0.05). In addition, the collagen content, MVD, and the expression of VEGF and PDGF-BB in the ICA+rASCs group were significantly higher than those in rASCs group(P<0.05). Conclusion ICA can enhance the expression of angiogenesis factors in rASCs, which may be the mechanism of ICA optimizing rASCs to promote the survival of ultra-long random skin flaps in rat models. |
| Key words: Adipose-derived stem cells Icarin Ultra-long random flap Angiogenesis |
