人工真皮复合组织瓣修复对断指再植大鼠皮瓣成活及炎症反应、细胞凋亡的影响

Effects of artificial dermal composite tissue flap on flap survival,inflammatory response and apoptosis of severed finger replantation rats

目的研究人工真皮复合组织瓣修复对断指再植大鼠皮瓣成活及炎症反应、细胞凋亡的影响。方法选择20只成年雄性SD大鼠,体重180~200 g,10周龄,按照随机数字表法分为对照组和实验组,各10只。切断股骨后内固定的方法建立断指再植模型,对照组进行组织瓣修复,实验组进行人工真皮复合组织瓣修复。观察皮瓣存活率及存活面积;取皮瓣并采用蛋白质印迹检测核因子-κB(NF-κB)、NOD样受体热蛋白结构域相关蛋白3(NLRP3)、B淋巴细胞瘤-2基因(Bcl-2)、Bcl-2相关x蛋白(Bax)、裂解型含半胱氨酸的天冬氨酸蛋白水解酶-3(Cleaved caspase-3)的表达水平,TUNEL法检测细胞凋亡率,酶联免疫吸附试验检测肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β、IL-18的含量。结果实验组大鼠的皮瓣存活率及存活面积、皮瓣中Bcl-2的表达水平为(85.12±11.31)%、(7.41±1.25)cm 2、0.94±0.16,均高于对照组[(64.59±8.39)%、(5.75±0.95)cm 2、0.29±0.06],皮瓣中NF-κB、NLRP3、Bax、Cleaved caspase-3的表达水平、TNF-α、IL-1β、IL-18的含量、细胞凋亡率为0.33±0.07、0.47±0.0、0.67±0.14、0.34±0.07、(4.54±0.95)ng/mg、(4.09±0.78)ng/mg、(67.62±11.34)pg/mg、(7.55±1.58)%,均低于对照组[0.83±0.12、0.95±0.15、1.05±0.18、1.05±0.18、(7.52±1.15)ng/mg、(6.26±1.09)ng/mg、(93.49±13.85)pg/mg、(12.12±3.52)%],差异均有统计学意义(P<0.05)。结论人工真皮复合组织瓣修复用于断指再植大鼠能够促进皮瓣成活、抑制炎症反应及细胞凋亡。

Objective To study the effects of artificial dermal composite tissue flap on the flap survival,inflammatory response and apoptosis of severed finger replantation rats.Methods Twenty adult male SD rats,weighing 180~200 g,10 weeks old were selected and divided into control group and experimental group according to the random number table method,each 10 rats.The severed finger replantation model was established by internal fixation after cutting off the femur.The control group was replanted with tissue flap,and the experimental group was replanted with artificial dermal composite tissue flap.The survival rate and survival area of the flap were observed.Then the flap was taken and nuclear factor-κB(NF-κB),Nod-like Receptor,Pyrin Domain Containing 3(NLRP3),B lymphocytoma-2 gene(Bcl-2),bcl-2 associated X protein(Bax)and cleaved caspase-3 were detected by western blot,the contents of tumor necrosis factor-α(TNF-α),interleukin(IL)-1β,IL-18 was detected by enzyme-linked immunosorbent assay,the apoptosis rate was detected by TUNEL.Results The flap survival rate and survival area,and the level of Bcl-2 expression in the flap of rats in the the experimental group were(85.12±11.31)%,(7.41±1.25)cm 2,0.94±0.16,which were higher than those in the control group[(64.59±8.39)%,(5.75±0.95)cm 2,0.29±0.06],the level of NF-κB,NLRP3,Bax,Cleaved caspase-3 expression in the flap,the content of TNF-α,IL-1β,and IL-18,the apoptotic rate were 0.33±0.07,0.47±0.0,0.67±0.14,0.34±0.07,(4.54±0.95)ng/mg,(4.09±0.78)ng/mg,(67.62±11.34)pg/mg,(7.55±1.58)%,which were lower than those in the control group[0.83±0.12,0.95±0.15,1.05±0.18,1.05±0.18,(7.52±1.15)ng/mg,(6.26±1.09)ng/mg,(93.49±13.85)pg/mg,(12.12±3.52)%],the differences were statistically significant(P<0.05).Conclusion Artificial dermal composite tissue flap can promote the survival of flap,inhibit inflammatory reaction and cell apoptosis in rats with severed finger replantation.