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目的探讨LPA_2在PC12细胞缺血再灌注损伤致细胞凋亡中发挥的作用。方法测最适缺氧时间:将PC12细胞分为6组,分别在三气培养箱中糖氧剥夺0、3、6、9、12、15 h。换高糖培养基普通培养箱中复氧24 h,MTT测细胞存活率;将PC12细胞分为5组:正常组、缺血再灌注组(OGD组)、缺血再灌注+溶剂对照组(OGD+DMSO组)、缺血再灌注+LPA_2激动剂(Dodecylphosphate)组(OGD+激动剂组)、缺血再灌注+LPA_2抑制剂(H2L5186303)组(OGD+抑制剂组),缺氧最适时间后再复氧24 h,MTT测细胞存活率,Western Blotting检测LPA_2及p-akt蛋白表达水平。结果缺氧处理12 h是模拟缺血再灌注损伤的最适时间;缺血再灌注组与正常组比较LPA_2表达水平降低;激动剂组与溶剂对照组比较LPA2表达水平增高,p-akt表达水平增高。结论升高LPA_2表达水平可提高细胞的存活率,LPA_2在缺血再灌注损伤中发挥保护作用。
Objective To investigate the role of LPA_2 in the apoptosis of PC12 cells induced by ischemia-reperfusion injury. Methods The optimal hypoxia time: the PC12 cells were divided into 6 groups, respectively, in a three-gas incubator oxygen and oxygen deprived 0,3,6,9,12,15 h. The cells were cultured for 24 hours in reoxygenation medium with high glucose medium for 24 h, MTT cell viability was measured. PC12 cells were divided into 5 groups: normal group, ischemia reperfusion group (OGD group), ischemia reperfusion + solvent control group OGD + DMSO group), ischemia-reperfusion + LPA2 (OGD + agonist group), ischemia reperfusion + LPA2 inhibitor (H2L5186303) After reoxygenation for 24 h, cell viability was measured by MTT assay. The expression of LPA 2 and p-akt protein was detected by Western Blotting. Results The hypoxia for 12 h was the best time to simulate ischemia-reperfusion injury. The expression of LPA 2 in ischemia-reperfusion group was lower than that in normal group. The expression of LPA2 was increased in agonist group and solvent control group, and the expression of p-akt Increase. Conclusions Increasing the expression level of LPA 2 can increase the survival rate of cells, and LPA 2 can play a protective role in ischemia / reperfusion injury.