Triton X-100与SDS对SD大鼠皮下移植猪主动脉瓣巨噬细胞表型极化的作用比较

目的:比较非离子型去污剂聚乙二醇辛基苯基醚(Triton X-100)和离子型去污剂十二烷基硫酸钠(SDS)对SD大鼠皮下移植猪主动脉瓣巨噬细胞表型极化的作用。方法:分别采用Triton X-100和SDS对猪主动脉瓣进行去细胞处理,将瓣膜分为Triton X-100处理组和SDS处理组,三羟甲基氨基甲烷(Tris)缓冲液处理组作为未处理组。将处理后的猪主动脉瓣埋入SD大鼠胸部左右两侧皮下,建立SD大鼠皮下移植猪主动脉瓣模型,分别在第3、14、28天时取出移植的猪主动脉瓣,免疫组织化学染色比较猪主动脉瓣中CCR7(M1型巨噬细胞标志物)和CD163(M2型巨噬细胞标志物)的表达情况,计算M2型巨噬细胞与M1型巨噬细胞的比值(M2型/M1型巨噬细胞=CD163+细胞数/CCR7+细胞数)。结果:未处理组猪主动脉瓣皮下移植28 d时,M2型/M1型巨噬细胞比值<1,巨噬细胞向M1型巨噬细胞极化,发挥促炎作用;SDS处理组、Triton X-100处理组瓣膜在皮下移植14 d和28 d时,M2型/M1型巨噬细胞比值>1,巨噬细胞向M2型巨噬细胞极化,发挥促进组织修复的作用。瓣膜皮下包埋14 d时,SDS处理组猪主动脉瓣M2型/M1型巨噬细胞比值明显高于Triton X-100处理组和未处理组(P均<0.05)。结论:去细胞处理可促进移植瓣膜中巨噬细胞向M2型巨噬细胞极化,采用SDS进行去细胞处理对巨噬细胞极化的作用优于Triton X-100。

Effects of Triton X-100 and SDS on phenotypic polarization of macrophages in SD rats subcutaneous transplanted porcine aortic valve

Objective:To compare the effects of non-ionic detergent polyethylene glycol octylphenyl ether(Triton X-100)and ionic detergent sodium dodecyl sulfate(SDS)on phenotypic polarization of macrophages in SD rats subcutaneous transplanted porcine aortic valve. Methods:The porcine aortic valve was decellularised by Triton X-100 and SDS separately and divided into Triton X-100 treatment group and SDS treatment group,while Tris buffer treatment group was used as blank control.After the porcine aortic valve was implanted into the left and right sides of SD rat′s chest to establish a subcutaneous transplantation of porcine aortic valve model in SD rats,the transplanted porcine aortic valve was removed at the 3 d,14 d and28 d respectively.Then the expression of CCR7(M1 macrophage marker)and CD163 (M2 macrophage marker)in the porcine aortic valve were compared by immunohistochemical staining and the ratio of CD163^+ cells/CCR7^+ cells(M2/M1 macrophage ratio) was calculated. Results:The results of immunohistochemistry showed that the M2/M1 macrophage ratio<1 in the blank control group at the 28th day of subcutaneous transplantation,indicating that non-cellular porcine valve tissue promoted polarization of macrophages into M1 macrophages in vivo and played an important role in promoting inflammatory response.M2/M1 macrophage ratio >1 in SDS and Triton X-100 treating valves at 14 d and 28 d after subcutaneous transplantation,suggesting that non-cellular porcine valve tissue promoted polarization of macrophages into M2 macrophages in vivo and played an important role in promoting tissue repair.The M2/M1 macrophage ratio of SDS group was significantly higher than those of Triton X-100 group and blank control group(both P <0.05). Conclusions:Decellularization can promote the polarization of macrophages into M2 macrophages in transplanted valves and SDS is superior to Triton X-100.