胰激肽原酶对他克莫司诱导肾损伤的保护作用

Protective effect of pancreatic kininogenase on tacrolimus-induced renal injury

目的研究胰激肽原酶(PK)在临床应用中对他克莫司(TAC)诱导的肾小管上皮细胞的保护作用及其机制。方法采用体外培养的方式将HK-2细胞划分为PK(6 pg/mL)组、TAC(50 ug/mL)组、对照组(CON组)、TAC(50 ug/mL)+PK(6 pg/mL)组。采用CCK-8检测每组细胞情况测定细胞活力;通过二氯二氢荧光素双醋酸盐(DCFH-DA)荧光与流式细胞法的组合应用,对细胞内部的活性氧分子(ROS)活跃度和数量进行检测;采用免疫印迹法对Bcl-2、Bax、LC3B、Beclin蛋白水平进行检测。结果PK组细胞活力显著低于CON组,TAC+PK组的细胞活力显著高于TAC组(P<0.05)。TAC组的细胞ROS表达能力要显著高于CON组;相较于TAC组,TAC+PK组细胞ROS表达能力略低(P<0.05)。TAC组的细胞自噬功能高于CON组,且细胞的LC3B-Ⅱ、Beclin表达能力较强;TAC+PK组的细胞LC3B-Ⅱ、Beclin表达能力要低于TAC组(P<0.05)。TAC组细胞的死亡数量大于CON组,Bcl-2/Bax比值下降;TAC+PK组Bcl-2/Bax比值显著大于TAC组(P<0.05)。结论胰激肽原酶对他克莫司诱导肾小管上皮细胞具有保护作用,其机制与胰激肽原酶抑制抗氧化应激及调节自噬有关。

Objective To study the pancreatic excitation peptide enzyme(pancreatic kallikrein,PK)in the clinical application of tacrolimus(tacrolimus,TAC)on the renal tubular epithelial cells induced by protective effect and its mechanism.Methods HK 2 cells were divided into pK(6 pg/mL)group,TAC(50 UG/mL)group,control group(CON group),TAC(50 UG/mL)+pK(6 pg/mL)group.CCK 8 was used to detect the condition of cells in each group and to determine the cell viability;The activity and quantity of reactive oxygen species(ROS)in cells were detected by the combination of dichlorodihydrofluorescein diacetate(DCFH DA)fluorescence and flow cytometry.The levels of bcl-2,Bax,lc3b and Beclin were detected by Western blot.Results The cell viability of PK group was significantly lower than that of con group,and the cell viability of TAC+PK group was significantly higher than that of TAC group(P<0.05).The expression of ROS in TAC group was significantly higher than that in con group.Compared with TAC group,the expression of ROS in TAC+PK group was slightly lower(P<0.05).The autophagy function of TAC group was higher than that of con group,and the expression of lc3bⅡand Beclin was stronger.The expression of lc3bⅡand Beclin in TAC+PK group was lower than that in TAC group(P<0.05).The number of cell death in TAC group was greater than that in con group,and the ratio of bcl-2/Bax decreased.The ratio of bcl-2/Bax in TAC+PK group was significantly higher than that in TAC group(P<0.05).Conclusion Pancreatic Kallidinogenase has a protective effect on renal tubular epithelial cells induced by tacrolimus.Its mechanism is related to the inhibition of antioxidant stress and the regulation of autophagy by Pancreatic Kallidinogenase.