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目的观察甘草甜素(GL)及地塞米松(DEX)对受损足细胞凋亡的影响,探讨其细胞学作用机制。方法建立嘌呤霉素(PAN)致足细胞损伤模型,应用不同水平PAN(12.5 mg.L-1、25.0 mg.L-1、50.0 mg.L-1、75.0 mg.L-1、100.0 mg.L-1)作用于足细胞,培养48 h后检测细胞凋亡率,选取合适的PAN作用质量浓度。将体外培养小鼠足细胞(MPC5)分为对照组、PAN组、DEX1组、DEX2组、DEX3组、GL1组、GL2组、GL3组,对照组加入等体积RPMI 1640培养液培养;PAN组加入PAN,终质量浓度50 mg.L-1;DEX1、2、3组同时加入PAN(终质量浓度50 mg.L-1)和DEX(终浓度分别为0.1μmol.L-1、1.0μmol.L-1、10.0μmol.L-1);GL1、2、3组同时加入PAN(终质量浓度50.0 mg.L-1)和GL(终质量浓度分别为400 mg.L-1、600 mg.L-1、800 mg.L-1),培养48 h。流式细胞仪检测各组细胞凋亡率。结果 50.0 mg.L-1 PAN诱导足细胞凋亡较对照组明显升高(P<0.05),75.0 mg.L-1、100.0 mg.L-1PAN诱导足细胞凋亡率显著升高,大部分足细胞死亡(Pa<0.01)。DEX1组、2组足细胞凋亡率明显低于PAN组(Pa<0.05),DEX 3组(10.0μmol.L-1)干预后足细胞凋亡率显著低于PAN组(P<0.01)。不同质量浓度GL组干预后足细胞凋亡率均显著低于PAN组(Pa<0.01),随GL质量浓度升高,其抗足细胞凋亡作用降低(P<0.05)。结论 PAN可呈剂量依赖性诱导足细胞损伤,50.0 mg.L-1 PAN为诱导合适的质量浓度。GL及DEX对PAN诱导足细胞损伤均有保护作用,DEX呈剂量依赖方式抑制足细胞凋亡,GL发挥抗足细胞凋亡作用与剂量有关。
Objective To observe the effect of glycyrrhizin (GL) and dexamethasone (DEX) on the apoptosis of injured podocytes and to explore their cytological mechanism. Methods To establish a model of puromycin (PAN) -induced podocyte injury, different levels of PAN (12.5 mg.L-1, 25.0 mg.L-1, 50.0 mg.L-1, 75.0 mg.L-1, 100.0 mg. L-1) on podocytes. After 48 h of culture, the apoptosis rate was detected and the appropriate concentration of PAN was selected. The cultured mouse podocytes (MPC5) were divided into control group, PAN group, DEX1 group, DEX2 group, DEX3 group, GL1 group, GL2 group, GL3 group and control group with equal volume of RPMI 1640 medium; PAN, the final concentration of 50 mg.L-1; DEX1,2,3 group were added PAN (final concentration of 50 mg.L-1) and DEX (final concentrations were 0.1μmol.L-1, 1.0μmol.L (10, 10μmol.L-1); GL1,2,3 group were added simultaneously PAN (final concentration of 50.0 mg.L-1) and GL (final concentration of 400 mg.L-1,600 mg.L -1,800 mg.L-1), cultured for 48 h. Flow cytometry was used to detect the apoptosis rate in each group. Results The apoptosis of podocytes induced by 50.0 mg.L-1 PAN was significantly higher than that of the control group (P <0.05). The apoptosis rate of podocytes induced by 75.0 mg.L-1, 100.0 mg.L-1PAN was significantly increased Podocyte death (Pa <0.01). The apoptosis rate of podocytes in DEX1 and 2 groups was significantly lower than that in PAN group (P <0.05). The apoptosis rate of podocytes in DEX1 group was significantly lower than that in PAN group (P <0.01) after intervention with DEX3 (10.0μmol.L-1). The apoptosis rate of podocytes in GL group was significantly lower than that in PAN group (P <0.01), and the anti-podocyte apoptosis was decreased with the increase of GL concentration (P <0.05). Conclusion PAN can induce podocyte injury in a dose-dependent manner, and 50.0 mg.L-1 PAN is suitable for inducing mass concentration. GL and DEX had protective effects on PAN-induced podocyte injury, DEX inhibited podocyte apoptosis in a dose-dependent manner, and GL exerted anti-podocyte apoptosis related to dose.