Department of Plastic Surgery, Peking Union Medical College Hospital, Beijing 100005, China.
Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100005, China.
Chin Med J (Engl). 2019 Jan 5;132(1):42-50. doi: 10.1097/CM9.0000000000000005.
Necroptosis is a new form of cell death that has been identified as a third pathway causing cell death. In this study, necrostatin-1 (Nec-1) was used to determine whether necroptosis exists in a rat ischaemia/reperfusion injury flap model.
In this study, twenty male Sprague-Dawley rats were divided randomly into two groups: a control group (CTL group) and a Nec-1 group. Each abdominal skin flap underwent 3 h of ischaemia and then reperfusion. Fifteen minutes before and after reperfusion, phosphate buffer saline (PBS) was administered intraperitoneally to the CTL group, while Nec-1 was administered intraperitoneally to the Nec-1 group. Twenty-four hours after reperfusion, the whole flap was divided equally into 54 sections. Flap blood perfusion was measured. One sample was taken randomly from each row. Morphological changes, apoptosis, receptor-interacting protein-1 (RIP-1) expression and caspase-3 activity were observed and detected. The measurements between the two groups were compared with the independent t-test, and a P value of <0.05 was considered statistically significant.
Compared to flaps in the CTL group, flaps in the Nec-1 group showed longer survival rates, better blood perfusion and less inflammatory infiltration. The total flap area considered to have survived was 70.88 ± 10.28% in the CTL group, whereas 80.56 ± 5.40% of the area was found to be living in the Nec-1 group (Nec-1 vs. CTL, t = -2.624, P < 0.05). For some rows, there were significant differences in cell apoptosis between the two groups, the apoptosis index (AI) in rows "9 cm", "7 cm", "6 cm" and "5 cm" was significantly lower in the Nec-1 group than that in the CTL group (Nec-1 vs. CTL, P < 0.05). RIP-1 expression was much lower in the Nec-1 group than that in the CTL group in rows "5 cm" to "9 cm" (Nec-1 vs. CTL, P < 0.05). No significant differences in caspase-3 activity were found.
According to the results, necroptosis was present in a rat abdominal ischaemia/reperfusion injury flap model.
坏死性凋亡是一种新的细胞死亡形式，已被确定为导致细胞死亡的第三种途径。在这项研究中，使用坏死抑制剂-1（Nec-1）来确定大鼠缺血/再灌注损伤皮瓣模型中是否存在坏死性凋亡。
在这项研究中，将 20 只雄性 Sprague-Dawley 大鼠随机分为两组：对照组（CTL 组）和 Nec-1 组。每个腹部皮瓣经历 3 小时的缺血，然后再灌注。在再灌注前 15 分钟和再灌注后，分别向 CTL 组腹腔内给予磷酸盐缓冲盐水（PBS），而向 Nec-1 组腹腔内给予 Nec-1。再灌注后 24 小时，将整个皮瓣均等分为 54 个部分。测量皮瓣的血流灌注。从每一行中随机抽取一个样本。观察并检测形态学变化、细胞凋亡、受体相互作用蛋白-1（RIP-1）表达和半胱天冬酶-3 活性。两组之间的测量值采用独立 t 检验进行比较，P 值<0.05 认为具有统计学意义。
与 CTL 组皮瓣相比，Nec-1 组皮瓣的存活率更高，血流灌注更好，炎症浸润更少。CTL 组总皮瓣面积被认为存活的有 70.88±10.28%，而 Nec-1 组有 80.56±5.40%的面积被认为存活（Nec-1 与 CTL，t=-2.624，P<0.05）。对于某些行，两组之间的细胞凋亡存在显著差异，Nec-1 组的凋亡指数（AI）在行“9 cm”、“7 cm”、“6 cm”和“5 cm”显著低于 CTL 组（Nec-1 与 CTL，P<0.05）。Nec-1 组 RIP-1 表达在行“5 cm”至“9 cm”的表达均明显低于 CTL 组（Nec-1 与 CTL，P<0.05）。半胱天冬酶-3 活性无显著差异。
根据结果，大鼠腹部缺血/再灌注损伤皮瓣模型中存在坏死性凋亡。
