右美托咪定对大鼠肋间后动脉穿支皮瓣缺血再灌注损伤保护作用的初步研究

doi:10.13591/j.cnki.kqyx.2022.05.002

摘要/Abstract

摘要： 目的探讨右美托咪定对大鼠背部肋间后动脉穿支皮瓣缺血再灌注损伤的保护作用。方法将24只SD大鼠随机分成空白对照组(对照组)、缺血再灌注组(I/R组)、缺血再灌注右美托咪定干预组(DEX+I/R组)。对照组进行大鼠肋间后动脉穿支皮瓣的制备,I/R组制备皮瓣后予以血管蒂夹闭6 h,DEX+I/R组在进行前述缺血处理后予以右美托咪定100 μg/kg干预并于24 h后重复给药。分组手术7 d后,通过大体观察判断皮瓣成活率,以HE染色观察大鼠皮瓣组织病理情况,采用TUNEL荧光染色检测各组大鼠皮瓣组织细胞凋亡情况,采用酶联免疫吸附法检测大鼠血浆中SOD、MDA含量,采用蛋白免疫印迹法检测大鼠皮瓣组织中凋亡相关蛋白(C-caspase-3、Bcl-2)、PI3K/Akt通路蛋白(PI3K、pAkt/Akt)表达情况。结果对照组、I/R组及DEX+I/R组大鼠皮瓣的成活率分别为(94.15±2.28)%、(38.15±10.48)%、(57.71±10.22)%。I/R组皮瓣成活率相较于与对照组显著降低(P＜0.001);DEX+I/R组相较于I/R组皮瓣成活率显著提高(P=0.006)。在距离血管蒂1 cm处,I/R组皮瓣组织细胞凋亡率相较于对照组升高(P＜0.001);DEX+I/R组中皮瓣组织细胞凋亡率较I/R组降低(P＜0.001)。在距离血管蒂2 cm处,I/R组皮瓣组织细胞凋亡率相较于对照组升高(P＜0.001);DEX+I/R组中皮瓣组织细胞凋亡率较I/R组降低(P=0.004)。I/R组大鼠血清SOD活力相较于对照组降低(P＜0.001);与I/R组比较,DEX+I/R组大鼠血清中SOD活力升高(P＜0.001);I/R组大鼠血清MDA含量相较于对照组升高(P<0.001);与I/R组相比,DEX+I/R组大鼠血清中MDA含量降低(P＜0.001)。I/R组中Bcl-2(P=0.001)、PI3K(P=0.003)、Akt(P=0.023)及pAkt(P=0.002)的蛋白相对表达量相较于对照组明显下降,而C-caspase-3的相对表达量明显上升(P=0.002);DEX+I/R组Bcl-2(P=0.003)、PI3K(P＜0.001)、Akt(P＜0.001)及pAkt(P＜0.001)的蛋白相对表达量相较于I/R组明显增高,而C-caspase-3的表达量下降(P＜0.001)。结论右美托咪定对大鼠背部肋间后动脉穿支皮瓣缺血再灌注损伤具有保护作用。

关键词: 肋间后动脉穿支皮瓣, 缺血再灌注损伤, 右美托咪定

Abstract: Objective To probe into the protective effect of dexmedetomidine on ischemia-reperfusion injury of posterior intercostal artery perforator flap in rats. Methods Twenty-four SD rats were randomly divided into control group, Ischemia-reperfusion group (I/R group) and dexmedetomidine+Ischemia-reperfusion intervention group (DEX+I/R group). Rats in control group was prepared with the posterior intercostal artery perforator flap; rats in I/R group were prepared with vascular pedicle clipping for 6 hours after flap harvesting, and the DEX+I/R group was treated with dexmedetomidine 100 μg/kg after ischemia process and repeated dosing 24 hours later. On day 7 after flap construction, survival rate of the flap was determined by gross observation; HE staining was used to observe the pathological condition of rat skin flap tissue; TUNEL fluorescence staining was used to detect the apoptosis of rat skin flap tissue; contents of SOD and MDA in rat plasma were determined by enzyme-linked immunosorbent assay. The expressions of apoptosis-related proteins (C-caspase-3, Bcl-2) and PI3K/Akt pathway proteins (PI3K, pAkt/Akt) in rat skin flap tissues were detected by western blotting. Results Survival rate of the flap in control group, I/R group and DEX+I/R group was (94.15±2.28)%, (38.15±10.48)% and (57.71±10.22)%. Compared with control group, the flap survival rate in I/R group was significantly lower (P＜0.001); compared with I/R group, the flap survival rate in DEX+I/R group was significantly higher (P=0.006). At the distance of 1cm from vascular pedicle, the apoptosis rate of skin flap tissue in I/R group was higher than that in control group (P＜ 0.001); the apoptosis rate of skin flap tissue in DEX+I/R group was lower than that in I/R group (P＜ 0.001). At the distance of 2 cm from vascular pedicle, the apoptosis rate of skin flap tissue in I/R group was higher than that in control group (P＜0.001); the apoptosis rate of flap tissue in DEX+I/R group was lower than that in I/R group (P=0.004). Compared with control group, SOD activity in serum of rats in I/R group was lower (P＜0.001); compared with I/R group, SOD activity in serum of DEX+I/R group was increased (P＜0.001). The serum MDA content in I/R group was higher than that in control group (P＜ 0.001); compared with I/R group, MDA content in serum of DEX+I/R group was decreased (P＜0.001). The expression of Bcl-2 (P=0.001), PI3K (P=0.003), Akt (P=0.023) and pAkt (P=0.002) in I/R group were lower compared with control group, while the expression of C-caspase-3 was significantly increased (P=0.002). The expression of Bcl-2(P=0.003), PI3K(P＜0.001), Akt(P＜0.001) and pAkt(P＜0.001) in I/R group were increased compared with control group, while the expression of C-caspase-3 was significantly decreased (P＜0.001). Conclusion Dexmedetomidine can alleviate ischemia-reperfusion injury of posterior intercostal artery perforator flap in rats.

Key words: posterior intercostal artery perforator flap, ischemia-reperfusion injury, dexmedetomidine

中图分类号:

R782.25

尹忠浩, 朱载瓯, 杨月美, 卢鹏, 宋晓萌, 吴煜农. 右美托咪定对大鼠肋间后动脉穿支皮瓣缺血再灌注损伤保护作用的初步研究[J]. 口腔医学, 2022, 42(5): 391-398.

YIN Zhonghao, ZHU Zaiou, YANG Yuemei, LU Peng, SONG Xiaomeng, WU Yunong. Protective effect of dexmedetomidine on ischemia-reperfusion injury of posterior intercostal artery perforator flap in rats[J]. Stomatology, 2022, 42(5): 391-398.

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