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阿托伐他汀钙影响大鼠背部超长随意皮瓣成活的研究

Effects of Atorvastatin Calcium on the Survival of Ultra-long Dorsal Random Skin Flaps in Rats

    摘要
  • 摘要:
      目的  探讨不同剂量阿托伐他汀钙（atorvastatin calcium, ATR）对大鼠背部超长随意皮瓣成活的影响。
      方法  将30只SD大鼠随机分为5组，每组6只：0.9%生理盐水对照组（CON组）、ATR（10 mg/kg）组（P10组）、ATR（20 mg/kg）组（P20组）、ATR（30 mg/kg）组（P30组）、ATR（40 mg/kg）组（P40组）。各组的每只大鼠背部制作8 cm×2 cm的随意皮瓣并原位回植。在术前3 d和术后3 d连续6 d按分组灌胃对应剂量的ATR或0.9%生理盐水，每天1次。术后第7 天，肉眼观察大体皮瓣外观，计算皮瓣的成活率并切取皮瓣存活区域组织（距坏死区0.5 cm的存活皮瓣），分别行HE染色观察皮瓣组织学变化，免疫组织化学法检测皮瓣微血管数量，实时荧光定量PCR检测血管内皮生长因子（vascular endothelial growth factor, VEGF）、碱性成纤维细胞生长因子（basic fibroblast growth factor, bFGF）mRNA的表达，酶联免疫吸附法测定（enzyme linked immunosorbent assay, ELISA）超氧化物歧化酶（superoxide dismutase, SOD）、一氧化氮（nitrogen monoxide, NO）、丙二醛（malonaldehyde, MDA）的含量。
      结果  术后第7天，大鼠大体可见CON组皮瓣出现大面积坏死，坏死部分形成痂壳，痂壳质硬，无弹性，并见大量组织液渗出，筋膜层组织呈暗紫色；P10、P20、P30、P40组皮瓣未见组织液渗出，痂壳自然脱落，皮毛正常生长。HE染色结果显示CON组皮瓣组织中出现大量炎症细胞浸润，伴有表皮缺失、坏死等病理性改变，但ATR治疗后皮瓣组织的病理性改变得到明显改善。与CON组比较，随著ATR剂量提高，皮瓣存活率，皮瓣微血管数量，皮瓣中VEGF mRNA、bFGF mRNA、SOD和NO水平也相应提高，在ATR 30 mg/kg达峰（P<0.05），但皮瓣中MDA水平却随ATR剂量提高而降低，在ATR 30 mg/kg最低（P<0.05）。
      结论  30 mg/kg的ATR对提高大鼠背部超长随意皮瓣的成活率效果最佳，这与ATR促进微血管生成、抗炎及抑制氧化应激相关。

     

    Abstract:
      Objective  To investigate the effects of atorvastatin calcium (ATR) on the survival of ultra-long dorsal random skin flaps in rats.
      Methods  Thirty SD rats were divided into five groups (n=6) according to the random number table: normal saline control group (CON group), ATR 10 mg/kg group (P10 group), ATR 20 mg/kg group (P20 group), ATR 30 mg/kg group (P30 group), and ATR 40 mg/kg group (P40 group). After pretreatment with ATR or 0.9% saline for 3 days, an ultra-long dorsal random skin flaps with size of 8 cm×2 cm was made on the back of each rat and replanted in situ. After the operation, the ATR or saline treatment lasted for 3 d. Seven days after operation, the appearance of skin flaps was observed with naked eyes, the survival rate of skin flaps was calculated. The pathological changes in the surviving areas of skin flaps were observed by HE staining, the number of microvessels in skin flaps was observed by immunohistochemistry staining, the mRNA expressions of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) were tested by quantitative real-time PCR, and the contents of superoxide dismutase, nitrogen monoxide and malonaldehyde were tested by enzyme linked immunosorbent assays (ELISA).
      Results  On the 7th day after operation, the skin flap of the CON group showed a large area of necrosis, and the necrotic part formed crusts. Crusts were hard and inelastic, and a large amount of tissue fluid exudated. The fascial layer showed dark purple. No exudation of tissue fluid was observed in the flaps of P10, P20, P30 and P40 groups. The scab shell fell off naturally and the fur grew normally. HE staining of CON group showed that a large number of inflammatory cell infiltration, epidermal loss and necrosis in skin flaps, but the pathological changes in skin flaps were significantly improved after treatment with ATR. Compared with the CON group, the survival rate of skin flaps, the number of microvessels in skin flaps and the levels of VEGF mRNA, bFGF mRNA, SOD, NO in skin flaps also increased with the dose of ATR, which reached a peak at 30 mg/kg ATR (P<0.05). However, the level of MDA in skin flaps decreased with the dose of ATR, which reached the lowest at 30 mg/kg ATR (P<0.05).
      Conclusion  ATR can enhance the survival of ultra-long dorsal random skin flaps in rats, which may be related to promoting microangiogenesis and inhibiting inflammatory and oxidative stress.

     

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