bFGF与MSCs自体移植对猪皮肤创面PCNA和Ⅷ-R Ag表达的影响

Effect of bFGF in coordination with autografting of bone marrow mesenchymal stem cells on expression of PCNA and Ⅷ-R Ag of porcine skin defect wound

目的研究碱性成纤维细胞生长因子(basicfibroblastgrowthfactor,bFGF)和骨髓间充质干细胞(mesenchy-malstemcells,MSCs)自体移植对小型猪全层皮肤缺损创面增殖细胞核抗原(PCNA)和Ⅷ因子相关抗原(Ⅷ-RAg)表达的影响。方法6只小型猪背部中线两侧各制备4个面积为2.54cm2的圆形全层皮肤缺损创面,将创面分为bFGF治疗组、MSCs治疗组、bFGF+MSCs治疗组、生理盐水对照组(共4组,每组12个创面)。分别取不同时段创面组织,进行常规染色,并采用免疫组化方法检测PCNA和Ⅷ-RAg的表达情况。结果伤后7d各组创面均形成肉芽组织,以bFGF+MSCs治疗组的血管形成最为丰富。PCNA表达量增加,伤后7d达到高峰,主要位于肉芽组织中,持续至14dPCNA表达开始下降,而且在各个时间点,均以bFGF+MSCs治疗组的PCNA阳性细胞百分率为最高(P<0.05)。伤后7d,Ⅷ-RAg染色的阳性细胞百分率明显高于其余3个组。结论bFGF协同自体移植的MSCs可使皮肤创面PCNA和Ⅷ-RAg的表达增加,促进肉芽组织生成,缩短创面愈合时间。

Objective To investigate the effect of bFGF and MSCs autografting on expression of PCNA and Ⅷ-R Ag of porcine skin defect wound. Methods Six male minipigs were used to produce 48 full-thickness skin wounds, each wound area about 2.54 cm^2 on both sides of the back （8 wounds per pig）. These 48 wounds were randomly treated with bFGF, MSCs, bFGF ＋ MSCs, and normal saline （ 12 wounds for each treatment）. The tissue from the wounds was harvested on day 3, 7, 14 after surgery, then used for HE staining and immu- nohistochemical staining for proliferating cell nuclear antigen （ PCNA ） and factor Ⅷ related antigen （ Ⅷ-R Ag）. Results Granulation tissue was presented in all groups on day 3 and 7 after wound, most abundant in the wounds treated with bFGF ＋ MSCs, as well as blood capillary formation. PCNA expression increased after skin injury, reached the peak on day 7, most in the granulation tissue, and lasted to day 14, then began to decrease. On day 3, 7, 14 after skin injury, PCNA expression was the most highest in the wounds treated with bFGF ＋ MSCs. Ⅷ-R Ag-positive cells in the wounds treated with bFGF ＋ MSCs were significantly more than that of the other three treatments （P 〈 0. 05）. Conclusion bFGF in coordination with MSCs can promote the expression of PCNA and Ⅷ-R Ag of wounds and accelerate granulation tissue growth.