硝酸盐转运蛋白Sialin与游离组织瓣缺血再灌注损伤的相关性研究

Research for relevance between nitrate transporters （Sialin） and ischemia-reperfusion injury in free flaps

目的探讨硝酸盐转运蛋白Sialin与游离组织瓣缺血再灌注损伤的相关性。方法2015年7月-2015年9月。将20只大鼠随机分为对照组和实验组．每组10只。在大鼠左下腹部制备大小4cmx8cm的游离组织瓣，实验组缺血6h后再灌注，对照组不进行缺血处理。组织瓣原位缝合。术后观察7d，于组织瓣成活范围最远端取样本。通过Real—timePCR、Western blot技术分析Sialin的表达水平，利用化学发光法检测技术分析硝酸盐含量的变化。结果采用Real—timePCR进行相对定量分析，实验组SialinmRNA表达水平为0．284±0．0486，显著低于对照组（0．945±0．0530）（P〈0．05）。应用蛋白印迹（Westernblot）技术分析，实验组Sialin蛋白表达水平为（0．1413±0．0446）。显著低于对照组（0．3519±0．0368）（P〈0．05）。硝酸盐浓度检测结果发现，硝酸盐转运蛋白Sialin与组织瓣硝酸盐浓度呈线性相关，Slain表达水平下降，硝酸盐浓度下降（P〈0．05，R=0．81）。结论硝酸盐浓度与Sialin表达水平呈线性相关，即Sialin表达水平下降，硝酸盐浓度下降，硝酸盐转运蛋白Sialin可能与组织瓣缺血再灌注损伤存在相关性。

Objective To explore the relevance between nitrate transporters （Sialin） and ischemia-reperfusion injury （IRI） in free flaps. Methods Twenty rats were randomly divided into control group and experimental group, 10 in each group. After preparatiing 4 cm×8 cm free tissue flap on left lower abdominal, the rats in experimental group were experienced ischemia in 6 hours, then reperfusion. The rats in control group were without ischemia treatment, and tissue flaps were sutured in situ. After postoperative observation for 7 days, the furthest edge of survival flaps were sampled. The expression of Sialin was detected by the Real-time PCR and Western blot. The change of nitrate content was analysed by chemiluminescence. Results Both of results of Real-time quantitative PCR and Western blot, it was found that expression levels of Sialin in the experimental group were significantly lower than the control group（0.945±0.0530）（P〈0.05）. Applying Real-time quantitative PCR relative quantitative analysis, it was found that the experimental group Sialin mRNA expression level was 0.284±0.0486, significantly lower than that of control group （0.945±0.0530） （P〈0.05）. Using Western blot technical analysis, experimental Sialin protein expression level was（0.1413±0.0446）, significantly lower than the control group （0.3519±0.0368） （P〈0.05）. The concentration of nitrate of test results found that the expression of nitrate trans- porters （Sialin） linearly related with the concentration of nitrate in free flaps, which means the Sialin expression level de- creased, the nitrate content decreased（P〈0.05, R=0.81）. Conclusion The concentration of nitrate was linearly related to the level of Sialin expression： when Sialin expression levels dropped, the concentration of nitrate decreased. Nitrate trans- porters（Sialin） may be correlated with IRI in free flaps.