锌对皮瓣缺血再灌注损伤细胞保护作用的研究

STUDY ON PROTECTIVE ROLES OF ZINC ON CELLS IN RAT FLAPS WITH ISCHEMIA-REPERFUSION INJURY

目的观察外源性锌在皮瓣缺血再灌注(ischem ia-reperfus ion,IR)损伤中的细胞保护作用,并探讨其机制。方法48只健康雄性W istar大鼠,制备以腹壁浅动脉为蒂的轴型皮瓣IR模型。大鼠随机分为对照组、IR组和补锌-IR实验组,每组16只。各组分别取8只大鼠,IR组和实验组于掀起皮瓣即刻、再灌注后1 h和24 h分别于皮瓣中段边缘取全层皮瓣组织1.0 cm×0.5 cm,对照组于掀起皮瓣即刻、9 h和32 h同法取材,分别行免疫组织化学检测金属硫蛋白(m eta lloth ione in,M T)的表达部位,并对切片进行图像分析;透射电镜观察IR后超微结构的改变和皮瓣成活率;生物化学检测皮瓣组织中丙二醛(m a lond ia ldehyde,M DA)含量和髓过氧化物酶(m ye loperox idase,M PO)活性。结果掀起皮瓣即刻对照组、IR组和实验组皮瓣M DA、M PO水平相近,差异无统计学意义(P>0.05)。缺血再灌注后1、24 h,IR组M DA含量分别高于对照组62.2%和136.4%(P<0.01),高于实验组11.3%和33.2%(P<0.05);M PO活性分别高于对照组238.4%和503.4%(P<0.01),高于实验组17.9%、24.1%(P<0.05)。掀起皮瓣即刻对照组和IR组M T含量极低,无法定量分析,实验组为45.30±7.60。缺血再灌注后1、24 h,实验组M T含量分别较IR组高41.5%和44.9%(P<0.01),IR组M T含量分别较对照组高119.9%和234.6%(P<0.01)。对照组皮瓣成活率为100%,IR组皮瓣成活率为19.65%±4.38%,实验组皮瓣成活率为24.99%±5.12%,较IR组提高27.2%(P<0.05)。结论外源性锌能对皮瓣内多种细胞的IR损伤产生保护作用,可明显提高皮瓣成活率。

Objective To observe the protective role of the ectogenesis zinc on the cells in rat flap with isehemia reperfusion injury and study the mechanisms. Methods A right low abdominal island flap was created in Wistar rats. Forty-eight rats were randomly divded into 3 groups （n--16） ：the control group, the isehemia reperfusion group and adding zinc isehemia reperfusion group. The content of malondialdehyde （MDA） and the activity of myeloperoxidase （MPO） were measured by thiobarbiturie acid methods and eolorimetry. The location of expression of MT was observed ,and the image analysis was performed. The quantity of MT was represented by the integratialoptical density. The ultrastrueture changes of skin flap with isehemia reperfusion injury and the flap viability were observed. Results In the isehemia reperfusion injury flaps, the content of MDA and MPO show no statistically significant difference among the control group, IR group and the adding-zine-IR group （P〉0.05）. Compared with the control group at 1 h and 24 h of reperfusion, the level of MDA increased 62.2% and 136.4%（P〈0.01） in the IR group, which increased 11. 3% and 33. 2%（P〈0. 01） in the adding zincdR group. The activity of MPO increased 238. 4% and 503.4% （P〈0.01） in the IR group when compared with the control group, and increased 17. 9% and 24.1%（P〈 0.05） when compared with the adding-zinc-IR group. In the ischemia reperfusion injury falps, the content of MT in the control group and the IR group is too minimal to measure. While the content of MT in the adding-zinc-IR group is 45.30± 7.60. At 1 h and 24 h of reperfusiion, the content of MT in the adding-zinc IR group increased 41. 5%o and 44.9% （P〈0.01） compared with the IR group, and increased 119. 9% and 234.6% （P〈0.01） compared with the control group. The flap viability is 100% in the control group, 19.65%±4.38% in the IR group, and 24.99%±5. 12% in the adding zinc IR group, which increased 27.2% （P〈0.05） compared with IR group. Conclusion Many kinds of cells in skin flap with ischemia reperfusion injury can be protected by ectogenesis zinc and the flap viability increases significantly.