保留蒂部筋膜对大鼠肋间后动脉穿支蒂皮瓣成活的影响
Effect of preserved pedicle fascia on the survival of rat posterior intercostal artery perforator pedicle flap
摘要目的:探讨保留蒂部筋膜对大鼠肋间后动脉穿支蒂皮瓣成活的影响。方法:50只SD大鼠,雌雄不限,采用随机数字表法分为A、B 2组,每组25只。于每只SD大鼠右侧背部设计并切取以肋间后动脉穿支为蒂的皮瓣,大小为10.0 cm×2.5 cm,A组保留穿支及其周围2.5 cm×1.0 cm的筋膜组织,B组仅保留穿支,尽可能剔除穿支周围的筋膜组织,最后均将皮瓣原位缝合。观察术后2组皮瓣成活情况,并计算皮瓣成活面积百分比;切取2组皮瓣组织行HE染色,于光镜下观察标本血管生成情况,并应用Image-Pro Plus 6.0软件统计术后第7天HE染色切片中的血管密度,测量并计算血管直径及血管壁厚度;应用免疫组织化学检测2组皮瓣组织中低氧诱导因子-1α(HIF-1α)和细胞珠蛋白(CYGB)表达的情况;蛋白质印迹法检测2组皮瓣组织中HIF-1α和CYGB的蛋白相对表达量,并比较2组间的差异。计量资料以 ± s表示,2组间数据比较采用双侧 t检验, P<0.05表示差异有统计学意义。 结果:A组皮瓣术后成活良好,B组术后7 d皮瓣远端缺血坏死,A组皮瓣成活面积百分比为96.19%±0.65%,高于B组的88.97%±3.24%,差异有统计学意义( P<0.01)。HE染色结果显示,2组皮瓣组织均可见新生血管,A组新生血管多于B组;A组血管密度、血管直径、血管壁厚度分别为(7.77±0.92)条/mm 2、(65.23±14.62) μm、(10.49±1.99) μm,均高于B组的(5.82±0.85)条/mm 2、(40.61±7.78) μm、(6.27±2.63) μm,差异均有统计学意义( P均<0.05)。免疫组织化学结果显示,术后第1天A组HIF-1α表达高于B组,并逐渐趋于稳定;A组和B组CYGB表达于术后第7天达到高峰。蛋白质印迹结果显示,术后第1、3天A组中HIF-1α表达量高于B组( P均<0.01),于术后第5天达到峰值后逐渐下降,并趋于稳定;术后第5、7、9天B组中HIF-1α表达量高于A组( P < 0.01),直至第9天出现缓慢下降的趋势。术后第3、5、9天A组中CYGB表达量均高于B组( P<0.01),A组和B组CYGB表达量均于术后第7天出现峰值。 结论:大鼠肋间后动脉穿支蒂皮瓣切取时在蒂部保留部分筋膜,可提高皮瓣成活面积,可能是其中的HIF-1α的表达稳定及CYGB的表达上调所致。
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abstractsObjective:To investigate the effect of preserved pedicle fascia on the survival of rat posterior intercostal artery perforator pedicle flap.Methods:Fifty SD rats (male and female) were randomly divided into two groups, A and B, with 25 rats in each group. The flap pedicled with the perforating branch of the posterior intercostal artery was designed and cut from the right back of each SD rat, with the size of 10.0 cm×2.5 cm. The perforator branch and the surrounding fascia tissue of 2.5 cm×1.0 cm were retained in group A, while only the perforator branch was retained in group B, the fascia tissue around the perforating branch was removed as much as possible. Finally, the two groups of flaps were sutured in situ. Flap survival of the two groups was observed and the survival rates were calculated. The two groups of flaps were cut and stained with HE, and the angiogenesis of the specimens was observed under a light microscope. Image-Pro Plus 6.0 software was used to calculate the blood vessel density in the sections stained with HE on the 7th day after surgery, vessel diameter and vessel wall thickness were measured and calculated. The expressions of hypoxic-inducible factor-1α (HIF-1α) and cytoglobin (CYGB) in the two groups of flaps were detected by immunohistochemistry. The relative expressions of HIF-1α and CYGB in the two groups were detected by Western blotting, and the differences between the two groups were compared. The measurement data were expressed as Mean±SD. Bilateral t-test was used to compare the data between the two groups, and P<0.05 indicated statistically significant difference. Results:The flaps in group A survived well, while the flaps in group B had distal ischemic necrosis. The survival rate of the flaps in group A (96.19%±0.65%) was higher than that in group B (88.97%±3.24%) at 7 days after operation, and the difference was statistically significant ( P<0.01). HE staining showed neovascularization in both groups. The new blood vessels in group A were higher than those in group B. Blood vessel density, blood vessel diameter and blood vessel wall thickness in group A were (7.77±0.92) vessels/mm 2, (65.23±14.62) μm and (10.49±1.99) μm, respectively, which were higher than those in group B [(5.82±0.85) vessels/mm 2, (40.61±7.78) μm and (6.27±2.63) μm]. The differences were statistically significant( P<0.05). Immunohistochemistry showed that HIF-1α expression in group A was higher than that in group B on day 1 after surgery, and gradually tended to be stable. CYGB expression in group A and Group B peaked on the 7th day after surgery. The results of Western blotting showed that the expression level of HIF-1α in group A was higher than that in group B on the first and third day after surgery, with statistical significance ( P<0.01). HIF-1α in group A reached its peak on the fifth day after surgery, then gradually decreased and tended to be stable. HIF-1α expression in group B was higher than that in group A on the 5th, 7th and 9th day after surgery ( P<0.01), and showed a slow decreasing trend until the 9th day. The expression level of CYGB in group A was higher than that in group B on the third, fifth and ninth day after surgery ( P<0.01), which had statistical significance. The expression of CYGB in group A and Group B reached a peak on day 7 after surgery. Conclusion:The survival rate of the rat posterior intercostal artery perforator flap can be improved by keeping part of fascia in the pedicle, which may be caused by the stable expression of HIF-1α and the upregulated expression of CYGB.
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