依达拉奉对氧剥夺引起的大鼠肠隐窝上皮细胞损伤的保护效应研究

Therapeutic efficacy of Edaravone on oxygen/glucose deprivation induced rat intestinal epithelial cells damage in vitro

关键词:  依达拉奉  氧剥夺  肠隐窝上皮细胞  凋亡

Key words:  Edaravone  oxygen/glucose deprivation  intestinal epithelial cells  apoptosis

摘要:

[目的] 研究依达拉奉对氧剥夺导致的IEC-6细胞株损伤的保护效应。 [方法] 将预先培养好的细胞分为3组：正常组(N组)，氧剥夺组(OGD，IR组)和依达拉奉治疗组(OGD+依达拉奉，E组)。本研究中所用氧剥夺模型是将细胞株换无糖无血清的培养基后放入含5% CO2和 95% N2的密闭培养箱中 2 h，后重新复氧复糖。在复氧复糖的同时，将依达拉奉盐溶液(10 mg/mL)加入细胞培养基中。在复氧复糖2 h，收集细胞用DHE荧光探针进行氧自由基(ROS)测定；伤后12 h，富集细胞用于超氧化物歧化酶(SOD)和丙二醛(MDA)的测定；伤后24 h收集细胞，用流式细胞仪测定细胞凋亡和 Alamar Blue 测定细胞活力。 [结果] DHE荧光探针结果显示OGD导致的ROS，E组比IR组减少；OGD刺激后，IEC-6细胞内的MDA浓度上升为（1.52±0.21） mmol/mg，而E组的含量为（1.21±0.16）mmol/mg，P<0.05。OGD后，IEC-6细胞内的SOD活力下降至（7.35±0.84）U/mg，而E组的SOD活力为（8.25±1.12）U/mg，P<0.05。OGD后细胞活力下降为0.48±0.081，而E组的细胞活力为0.56±0.049，P<0.01。同时，流式细胞仪的结果显示，依达拉奉抑制了OGD导致的细胞过度凋亡 。 [结论] 依达拉奉可以减轻OGD造成的IEC-6细胞的损伤。

Abstract:

[Objective] To investigate the protective effect of Edaravone on oxygen/glucose deprivation (OGD)-induced rat intestinal epithelial cells damage. [Methods] Cells were divided into normal group (with out OGD, group N), OGD group (OGD, group IR) and Edaravone treatment group (OGD+ edaravone treatment, group E). IEC-6 cells, cultured with serum and sugar free medium, were put in closed incubator filled with 5% CO2 and 95% N2 mixed gas and two hours latter, cells were changed to normal condition. Edaravone (10 mg/mL) were administered just after OGD treatment in group E. At 2 hs in normal condition, the cells were collected for ROS determination with the DHE probe. At 12 hs post injury, the cells were collected for detections of malonaldehyde (MDA) and superoxide dismutase (SOD). At 24 hs post injury, the cells were used for detection apoptosis with the flow cytometry and cell viability measurement with AlamarBlue. The DHE probe results showed that the ROS caused by OGD was less in goup E than IR. After OGD, MDA levels were increased to (1.52±0.21) mmol/mg and in Edaravone treatment group, the MDA levels were (1.21±0.16) mmol/mg (P<0.05). After OGD, SOD activities were decreased to (7.35±0.84) U/mg and in Edaravone treatment group, the SOD activities were (8.25±1.12) U/mg (P<0.05). After OGD, cell viability was decreased to 0.48±0.081 and in Edaravone treatment group, cell viability was 0.56±0.049 (P<0.01). Furthermore, the results from the flow cytometry showed that edaravone reduced the OGD-induced cell apoptosis. [Conclusion] Edaravone is an effective for OGD-induced IEC-6 cell damage.