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[Tempol对大鼠随意型皮瓣存活的影响]

[EFFECTS OF Tempol ON SURVIVAL OF RANDOM PATTERN SKIN FLAPS IN RATS].

作者信息

Li Guodong, Xu Yongqing, He Xiaoqing, Yang Fan, Chen Dewei, Luo Haotian, Yang Xi

机构信息

Brigade of Postgraduate Managementy, College of High Altitude Military Medicine, Third Military Medical University, Chongqing, 400038, P. R. China.

Department of Orthopedics, Kunming General Hospitaly, College of High Altitude Military Medicine, Third Military Medical University, Chongqing, 400038, P. R. China.

出版信息

Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2016 Oct 8;30(10):1264-1269. doi: 10.7507/1002-1892.20160258.

Abstract

OBJECTIVE

To study the effects of the new small molecular oxygen free radical scavenger Tempol on the survival and vasculogenesis of the long random pattern skin flap (LRPSF) and its mechanism.

METHODS

Eighty-four male Sprague Dawley rats were randomly divided into control and Tempol groups (42 rats in each group). LRPSF of 9 cm×3 cm in size were prepared on the backs of rats in two groups based on the Mcfarlane flap. Rats were administered with Tempol (100 mg/kg) in the Tempol group and with normal saline in the control group by intraperitoneal injection at 15 minutes before operation and at 1-7 day after operation. The rat and the skin flap survival conditions were observed after operation; the survival rate of skin flap was measured, and the vascular structure, vascular volume, and total length of blood vessels were analyzed with Micro-CT three-dimensional imaging after 7 days; HE staining was used to observe the structure of the skin flaps and inflammation, immumohistochemical staining to observe vascular endothelial growth factor (VEGF) expression; water-soluble tetrazolium-1 method was used to measure the content of superoxide dismutase (SOD) and malondialdehyde (MDA), and ELISA to detect the expressions of tumor necrosis factor α (TNF-α) and interleukin 6 (IL-6) after 1, 3, and 7 days.

RESULTS

All of rats survived after operation, without hemorrhage, edema, and infection. With the extension of time, necrosis occurred in the distal part of the skin flaps in 2 groups, but the necrosis degree of the Tempol group was lower than that of control group; meanwhile, the blood vessel distribution and continuity were better than those of control group. The skin flaps survival rate, vascular volume, and total length of blood vessels of Tempol group were significantly higher than those of control group after 7 days (<0.05). The clearer skin flaps structure, lighter inflammation reaction and inflammation cell infiltration, and higher VEGF staining intensity were observed in the Tempol group than the control group after 7 days. There was no significant difference in SOD, MDA, and TNF-α, and IL-6 contents between the 2 groups at immediate after operation. SOD significantly increased, but MDA, TNF-α, and IL-6 contents significantly decreased in the Tempol group when compared with control group after 1, 3, and 7 days (<0.05).

CONCLUSIONS

Tempol can significantly promote the LRPSF survival rates, its mechanism is closely related to the promotion of vasculogenesis and reduction of oxidative stress and inflammation.

摘要

目的

研究新型小分子氧自由基清除剂Tempol对长随意型皮瓣(LRPSF)存活及血管生成的影响及其机制。

方法

84只雄性Sprague Dawley大鼠随机分为对照组和Tempol组,每组42只。两组大鼠均参照Mcfarlane皮瓣在背部制备大小为9 cm×3 cm的LRPSF。Tempol组大鼠于术前15分钟及术后1~7天腹腔注射Tempol(100 mg/kg),对照组大鼠注射生理盐水。术后观察大鼠及皮瓣存活情况;测量皮瓣存活率,术后7天采用Micro-CT三维成像分析血管结构、血管容积及血管总长度;HE染色观察皮瓣结构及炎症情况,免疫组织化学染色观察血管内皮生长因子(VEGF)表达;采用水溶性四氮唑盐-1法检测术后1、3、7天超氧化物歧化酶(SOD)和丙二醛(MDA)含量,ELISA法检测肿瘤坏死因子α(TNF-α)和白细胞介素6(IL-6)表达。

结果

术后所有大鼠均存活,无出血、水肿及感染。随着时间延长,两组皮瓣远端均出现坏死,但Tempol组坏死程度低于对照组;同时,Tempol组血管分布及连续性优于对照组。术后7天,Tempol组皮瓣存活率、血管容积及血管总长度均显著高于对照组(<0.05)。术后7天,Tempol组皮瓣结构更清晰,炎症反应及炎症细胞浸润更轻,VEGF染色强度更高。术后即刻两组SOD、MDA、TNF-α及IL-6含量差异无统计学意义。术后1、3、7天,Tempol组SOD含量显著升高,MDA、TNF-α及IL-6含量显著降低,与对照组比较差异有统计学意义(<0.05)。

结论

Tempol可显著提高LRPSF存活率,其机制与促进血管生成、减轻氧化应激及炎症反应密切相关。

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