Effects of pretreatment with dimethyloxalylglycine on the survival of multi-territory perforator flap in rat and related mechanism
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摘要: 目的 观察二甲基乙二酰基甘氨酸(DMOG)预处理对大鼠跨区穿支皮瓣成活及闭塞区域2血管的影响,并探讨相关机制。 方法 取60只成年SD大鼠,按照随机数字表法分为DMOG组和生理盐水组,每组30只。在2组大鼠背部右侧制作三血管体穿支皮瓣,包括旋髂深动脉穿支、肋间动脉穿支、胸背动脉穿支和闭塞区域1、2。DMOG组大鼠于术前2 d、2 h及术后2 d腹腔注射2 mL含DMOG的生理盐水(DMOG剂量为40 mg/kg),生理盐水组大鼠于相同时相点腹腔注射等量生理盐水。术后7 d,行大体观察、计算皮瓣成活率;采用血管造影术观察皮瓣闭塞区域2及潜在区血管情况;HE染色观察皮瓣闭塞区域2的新生血管并计算微血管密度(MVD),免疫组织化学染色法和蛋白质印迹法检测该区域的血管内皮生长因子(VEGF)表达(分别以积分吸光度值与灰度值比值表示),激光多普勒血流成像仪检测该区域血管的血流量。每组各项指标的样本数均为6。对数据行
t 检验。 结果 (1)术后7 d,2组大鼠均存活,皮瓣未出现感染。DMOG组大鼠皮瓣的坏死区域大致位于胸背动脉穿支入皮点以远,痂皮呈暗黄色、质地较软;生理盐水组大鼠皮瓣的坏死区域大致位于闭塞区域2以远,痂皮呈棕黑色、质地较硬。DMOG组大鼠皮瓣成活率为(88±3)%,显著高于生理盐水组的(82±3)%(t =3.38,P <0.01)。(2)术后7 d,DMOG组大鼠皮瓣闭塞区域2的血管结构较清晰、新生血管较多,潜在区血管结构较完整;生理盐水组大鼠皮瓣闭塞区域2的血管结构模糊不清、新生血管较少,潜在区血管结构较紊乱。(3)术后7 d,DMOG组大鼠皮瓣闭塞区域2的MVD为(29.2±2.2)个/mm2,明显多于生理盐水组[(20.3±3.6)个/mm2,t =5.10,P <0.01]。(4)术后7 d,DMOG组大鼠皮瓣闭塞区域2 VEGF的免疫组织化学染色法和蛋白质印迹法表达结果分别为5 060±432、0.48±0.04,均明显高于生理盐水组(2 811±382、0.26±0.06,t 值分别为9.54、5.67,P 值均小于0.01)。(5)术后7 d,DMOG组大鼠皮瓣闭塞区域2血管的血流量为(58±4)灌注单位(PU),明显多于生理盐水组[(46±4)PU,t =5.20,P <0.01]。 结论 DMOG可通过促进大鼠背部跨区穿支皮瓣闭塞区域2的血管新生,改善皮瓣血供,从而提高皮瓣的成活率。-
关键词:
- 创伤和损伤 /
- 外科皮瓣 /
- 血管内皮生长因子类 /
- 闭塞血管 /
- 二甲基乙二酰基甘氨酸
Abstract: Objective To observe the effects of pretreatment with dimethyloxalylglycine (DMOG) on the survival of multi-territory perforator flap and the vessels of choke zone (CZ) 2 in rat, and to explore related mechanism. Methods Sixty adult SD rats were divided into group DMOG and normal saline group (NS) according to the random number table, with 30 rats in each group. Perforator flap with three angiosomes was made on the right dorsal side of rat, including deep iliac circumflex artery perforator, intercostal artery perforator, thoracodorsal artery perforator, as well as CZ 1 and CZ 2. Rats in group DMOG were intraperitoneally injected with 2 mL NS containing DMOG (40 mg/kg) 2 days before operation, 2 hours before operation, and 2 days after operation. Rats in group NS were intraperitoneally injected with equivalent volume of NS at the same time point. On post operation day (POD) 7, gross observation was conducted, and the survival rate of flap was calculated. On POD 7, the vascularity in CZ 2 and potential zone of flap was observed using angiography. On POD 7, new vessel in CZ 2 of flap was observed with HE staining, and the microvessel density (MVD) was calculated. On POD 7, the expression of vascular endothelial growth factor (VEGF) in CZ 2 of flap was detected by immunohistochemistry and Western blotting (respectively denoted as integral absorbance values and ratio of gray value), and blood flow volume of vessel in CZ 2 of flap was examined by laser Doppler perfusion imager. The sample number of each index was 6 in each group. Data were processed witht test. Results (1) On POD 7, rats in two groups all survived, and the flaps were not infected. In group DMOG, the necrotic area of flaps of rats with dark yellow crust and soft texture was observed approximately at the distal end of skin entry point of thoracodorsal artery perforator. In group NS, the necrotic area of flaps of rats with brownish black crust and hard texture was observed approximately at the distal end of CZ 2. The survival rate of flap of rats in group DMOG was (88±3) %, which was significantly higher than that in group NS [(82±3) %,t =3.38,P <0.01]. (2) On POD 7, there were clear vascular structure and many new vessels in CZ 2 of flaps of rats in group DMOG, with intact vascular structure in potential zone. On POD 7, there were unclear vascular structure and few new vessels in CZ 2 of flaps of rats in group NS, with disorder vascular structure in potential zone. (3) On POD 7, MVD in CZ 2 of flaps in rats of group DMOG was (29.2±2.2)/mm2, which was significantly higher than that of group NS [(20.3±3.6)/mm2,t =5.10,P <0.01]. (4) On POD 7, the expressions of VEGF in CZ 2 of flaps in rats of group DMOG detected by immunohistochemistry and Western blotting were 5 060±432 and 0.48±0.04 respectively, which were significantly higher than those of group NS (2 811±382 and 0.26±0.06, witht values respectively 9.54 and 5.67,P values below 0.01). (5) On POD 7, blood flow volume of vessel in CZ 2 of flaps in rats of group DMOG was (58±4) perfusion units (PU), which was significantly more than that of group NS [(46±4) PU,t =5.20,P <0.01]. Conclusions DMOG can increase the survival rate of multi-territory perforator flap through promoting angiogenesis in CZ 2 of flap on the back of rat and improving blood supply of flap.
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